Original Article
Prevalence and Spectrum of Germline Mutations of the p53 Gene among Patients with Sarcoma
N Engl J Med 1992; 326:1301-1308May 14, 1992
- Abstract
- Abstract
Background
Recent studies have identified germline mutations of the p53 tumor-suppressor gene in families with the Li—Fraumeni syndrome, a rare inherited disorder characterized by a high risk of sarcomas of bone and soft tissue, breast cancer, and other tumors. In this report, we address the possibility that some sporadic sarcomas may be associated with new germline mutations of the p53 gene, which would not be manifested as familial cancer unless the patient survived to reproduce.
Methods
We studied DNA from peripheral leukocytes of 196 patients with sarcoma and from 200 controls. Of the 196 patients with sarcoma, 15 were selected because they had had multiple primary cancers or had a family history of cancer. The entire coding sequence and splice junctions of the p53 gene were analyzed for mutations.
Results
Eight germline mutations were found, three in patients with no known family history of cancer and five in patients with an unusual personal or family history of cancer. Four mutations caused amino acid substitutions, and four caused stop codons. These mutations were not present in any of the 200 controls.
Conclusions
New germline mutations of the p53 gene are rare among patients with "sporadic" sarcoma but may be common in patients with sarcoma whose background includes either multiple primary cancers or a family history of cancer. Diverse mutations of this gene were associated with an increased likelihood of cancer; hence, the entire gene should be considered a target for heritable mutation. It appears that the group of patients with cancer who carry germline mutations of the p53 gene is more diverse than is suggested by the clinical definition of the Li—Fraumeni syndrome. The identification of carriers could be of substantial clinical importance. (N Engl J Med 1992;326: 1301–8.)
Article Activity
- Article
HEREDITARY mutations of tumor-suppressor genes have been correlated with a genetic predisposition to cancer, and several such genes have been linked to distinct familial cancer syndromes.1 The best-studied example is retinoblastoma; about 40 percent of the cases are due to germline mutations of the retinoblastoma gene (the remaining 60 percent of cases are nonhereditary and are due to somatic mutations).2 Patients with the hereditary form have a high risk of multifocal retinoblastoma and other tumors, and they may pass the disease to their progeny as an autosomal dominant trait. Three fourths of the hereditary cases of retinoblastoma occur in patients with no family history of the disease and are due to new mutations in the germline; the nonhereditary cases are unifocal, with an older average age of onset. If retinoblastoma is unifocal at the time of diagnosis, it is not always possible to differentiate the hereditary from the nonhereditary form of the disease.
Recent findings indicate that inheritance of a variant allele of the p53 gene may also predispose patients with the Li—Fraumeni syndrome to cancer, leading to a high risk of bone and soft-tissue sarcomas, breast cancer, and other neoplasms.3 , 4 Families with this syndrome are characterized by a proband with sarcoma and at least two other close relatives with sarcoma or carcinoma.5 The p53 gene product was first described as a cellular protein that forms a complex with large-T antigen in SV40-transformed cells.6 Recent studies have shown that the function of the wildtype p53 gene product is to control the normal cell cycle by regulating transcription7 , 8 or DNA replication,9 10 and other studies make it clear that the p53 gene is best classified, like the retinoblastoma gene, as a tumor suppressor.11 , 12 Somatic mutations of this gene have been found in many types of cancer, including all the major constituent tumors of the Li–Fraumeni syndrome.13 14 15 16
As compared with familial retinoblastoma, however, the Li—Fraumeni syndrome is relatively rare.6 , 17 One explanation could be that the spectrum of cancers associated with this syndrome is diverse, occurring early in life and in many cases having a poor prognosis.18 , 19 By contrast, the percentage of patients with retinoblastoma who survive into adulthood exceeds 90 percent.17 An alternative explanation could be that the Li—Fraumeni syndrome, as clinically defined, includes only a subgroup of the patients with cancer who have germline mutations of the p53 gene. This might be true if the clinical features associated with such mutations are very heterogeneous, depending on the specific mutation present or on other genetic or environmental factors. Studies have shown that both large deletions or insertions and small (point) mutations may be oncogenic in the p53 gene,20 , 21 but the majority of somatic point mutations cluster in five separate regions of the gene that were all highly conserved during evolution.14 , 22 The seven previously reported germline p53 mutations are even less diverse: all are amino acid substitutions occurring between codons 242 and 258.3 , 4 , 23
In this study, we have addressed several questions in an effort to define the role of the p53 gene in human cancers. First, we have examined the possibility that by analogy with retinoblastoma, some patients with sarcomas of bone and soft tissue and no family history of cancer may be carriers of new germline mutations at the p53 locus. This group might be indistinguishable from other patients with sarcomas at the time of the diagnosis of their first tumor, but they would carry a high risk of additional primary cancers during their lifetime and could pass this predisposition to their offspring. Second, we have examined the possibility that some patients with sarcoma whose personal or family history of cancer suggests a predisposition to cancer, but who do not necessarily fit the criteria for the Li—Fraumeni syndrome, carry germline mutations of the p53 gene. Finally, our study allows a direct comparison of the clinical manifestations of disease in two groups of patients with sarcoma: those who carry germline p53 gene mutations and those who do not. We have addressed these questions by screening 196 patients with sarcoma for germline mutations of the p53 gene using a combination of polymerase-chain-reaction (PCR) amplification of genomic DNA, analysis with single-strand conformation polymorphism (SSCP), and direct genomic sequencing. For comparison, we also analyzed 200 unrelated control subjects: 175 whose personal or family history of cancer was unknown and 25 with benign bone and soft-tissue tumors.
Methods
Subjects and DNA Samples
High-molecular-weight DNA was isolated from peripheral leukocytes of subjects as previously described.24 Peripheral-blood DNA samples were isolated from 196 patients with sarcoma; the majority (167) were of Japanese origin and living in Japan, whereas 29 were of mixed North American or Northern European descent and living in the United States or Australia. Among these patients the diagnosis was osteosarcoma in 95 cases; malignant fibrous histiocytoma in 26; chondrosarcoma in 21; synovial sarcoma in 15; liposarcoma in 8; rhabdomyosarcoma in 6; Ewing's sarcoma and malignant schwannoma in 5 each; chordoma in 3; fibrosarcoma, alveolar soft-parts sarcoma, leiomyosarcoma, and malignant hemangiopericytoma in 2 each; and epithelioid sarcoma, teratoma, malignant melanoma, and peripheral neuroepithelioma in 1 each. The patients were grouped in two ways. For the first group, 181 patients were randomly chosen from the patient population at several clinics in Japan and the United States without regard to whether they had a family or personal history of cancer. There were no other special criteria for inclusion in this group except for a diagnosis of sarcoma and a willingness to participate in a research study. For the second group, 15 patients with sarcoma were specifically selected as having had either multiple primary cancers or a family history of cancer. This group included seven patients who had multiple cancers thought likely to be of independent origin: three patients had multifocal osteosarcomas, one of whom had multifocal osteosarcoma after treatment for hepatoblastoma; three patients had a sarcoma in the field of irradiation used to treat primary breast cancer (two with malignant fibrous histiocytoma and one with synovial sarcoma); and one patient had osteosarcoma in the field of irradiation used to treat primary malignant fibrous histiocytoma. Among the remaining eight patients, three had a family history of cancer fitting the criteria for the Li—Fraumeni syndrome (that is, the proband was given a diagnosis of sarcoma before the age of 45 and had a first-degree relative with cancer in this age group and another first- or second-degree relative with cancer before the age of 45 or a sarcoma at any age)5 and five patients had a family history of cancer that could not be definitely classified as being part of the Li—Fraumeni syndrome (three patients had only one known relative with cancer, one patient had a grandmother with breast cancer and a niece with pinealoblastoma, and one patient had at least one relative with osteosarcoma but the exact number affected was unknown). The control group included 100 unrelated spouses of patients with a common hereditary (nonmalignant) disease, all of mixed North American descent, whose personal or family history of cancer was unknown; 75 unrelated adults of Japanese descent whose personal or family history of cancer was unknown; and 25 Japanese patients with benign bone or soft-tissue tumors.
Screening for Mutations
Initial screening for mutations was done by PCR amplification from genomic DNA followed by SSCP analysis of the amplified fragments.25 , 26 This approach has been shown to be a rapid and sensitive method for the detection of DNA sequence variations as small as a single-base substitution.25 DNA samples showing a variant band by SSCP analysis were analyzed by direct genomic sequencing to determine the underlying sequence variation.
SSCP Analysis
SSCP analysis was performed according to the procedure of Orita et al., with minor modifications.26 A separate pair of oligonucleotide primers was used to amplify each of the p53 exons 2 through 11 except for exon 5, in which two overlapping PCR fragments were analyzed. Each primer sequence was obtained from published data or by sequencing genomic clones of the human p53 gene provided by Dr. L. Crawford. PCR fragments included at least 10 base pairs (bp) of the 5' and 3' flanking regions of each exon. To limit the total length of the PCR fragments to less than 200 bp, thereby increasing the sensitivity of the SSCP technique, restriction-enzyme digestion was performed in some cases. PCR fragments were generated from 50 ng of genomic DNA in a 50-μl mixture containing 20 μmol of deoxyadenosine triphosphate, deoxythymidine triphosphate, or deoxyguanosine triphosphate per liter; 2 μmol of deoxycytidine triphosphate per liter; 1.0 to 2.5 mmol of magnesium chloride per liter; 20 pmol of each primer; 20 mmol of TRIS buffer per liter (pH 8.4 or 8.6); 50 mmol of potassium chloride per liter; 50 μg of bovine serum albumin per milliliter; 0.5 unit of Taq polymerase (Perkin—Elmer Cetus); and 0.1 μl (7 nmol per liter) of [α-32P]deoxycytidine triphosphate (3000 Ci per millimole). The reaction was performed for 30 cycles, consisting of 30 seconds of denaturation at 94°C, 90 seconds of annealing at 52 to 60°C, and 120 seconds of extension at 71°C with a programmable thermal controller (MJ Research). One tenth of the crude amplified product was mixed with 15 to 40 μl of a mixture of 0.1 percent sodium dodecyl sulfate and 10 mmol of EDTA per liter, followed by a 1:1 dilution with a loading solution consisting of 95 percent formamide, 89 mmol of TRIS buffer per liter, 2 mmol of EDTA per liter, 89 mmol of boric acid per liter, 0.05 percent bromophenol blue, and 0.05 percent xylene cyanol. Diluted samples were denatured at 95°C for two minutes, and then loaded on 6 percent nondenaturing polyacrylamide gels, once on a gel containing 10 percent glycerol and a second time on a gel that did not contain glycerol. Electrophoresis was carried out at room temperature for 8 to 16 hours at a constant power of 8 W. Autoradiography was performed for 12 to 24 hours without an intensifying screen.
Direct Genomic Sequencing
Direct genomic sequencing of double-stranded PCR fragments was performed according to protocols we have previously described.27 Briefly, amplified DNA samples were purified through Sepharose CL-6B (Pharmacia) and combined with 7 pmol of primers end-labeled with phosphorus-32. Mixtures of primers and template were heat-denatured, and sequencing reactions were carried out with the Sequenase enzyme (U.S. Biochemical). The product was analyzed by electrophoresis on 6 percent denaturing polyacrylamide gels. Autoradiography was carried out for 12 to 16 hours without an intensifying screen.
Results
Of the 196 patients with sarcoma we studied, 8 carried germline mutations of the p53 gene (Table 1Table 1
Germline Mutations of the p53 Gene in Eight Patients with Sarcoma.). Three germline mutations were identified in patients with sarcoma who had no family history of cancer at the time of diagnosis; the remaining five were found in patients who had a family history of cancer. Six of the 15 patients we selected for analysis on the basis of their unusual history of cancer (see the Methods section) had germline p53 mutations (3 of the 6 had a family history of cancer), whereas only 2 of the 181 randomly selected patients with sarcoma carried such mutations (both had a family history of cancer). Seven of the eight germline mutations were found in patients with a diagnosis of osteosarcoma, and one was found in a patient with malignant fibrous histiocytoma. In contrast to other investigators,3 , 4 , 23 we found that three mutations occurred outside the conserved domains of the p53 gene (codons 71 through 72, 151 through 152, and 209 through 210). The other five mutations were identified within the conserved domain (codons 120, 241, 245, 248, and 282). The types of mutations that we found were also heterogeneous: four caused amino acid substitutions, or missense mutations (codons 241, 245, 248, and 282); the remainder caused premature termination of protein translation, or nonsense mutations, through a single-base frame-shift mutation involving insertion (two cases), a two-base deletion (one case), or the direct creation of a stop codon (one case).None of the eight mutations found in the germline of patients with sarcoma were found in the control subjects.
Patients with No Unusual Family History of Cancer
Of the three patients with germline mutations and no unusual family history of cancer, the first was a patient with sarcoma who had two different primary tumors in her lifetime (Patient 1 in Table 1). At 17 years of age she was given a diagnosis of osteosarcoma on her right femur. Two years after treatment for this tumor, a focus of osteosarcoma was found on her right forearm. She was disease-free until the age of 28, when bilateral breast cancer was diagnosed. Orbital rhabdomyosarcoma developed in her daughter at the age of five years (Fig. 1Figure 1
Pedigree and Analysis of the New Germline Missense Mutation of the p53 Gene in the Family of Patient 1.A). Peripheral-leukocyte samples of DNA extracted from the patient and her daughter showed the same variant band on SSCP analysis of exon 7 of the p53 gene (Fig. 1B). The variant band was not present in any unaffected members of her family, including her parents (Fig. 1B). DNA sequence analysis showed that one p53 allele carried by this patient had a transition from guanine to adenine at the second nucleotide of p53 codon 248 (Fig. 1C). The mutation results in the substitution of a glutamine residue where arginine normally occurs in the p53 protein. Although we cannot exclude the possibility of mosaicism in either of her parents, it is reasonable to regard this mutation as a new germline mutation. Because she had no known family history of cancer and no personal history of cancer before osteosarcoma was diagnosed, she would have been regarded as having a "sporadic" case of osteosarcoma at presentation.The second patient also had two different primary tumors in her lifetime (Patient 2 in Table 1). Hepatoblastoma was diagnosed at the age of three months; treatment included radiation therapy. Multiple foci of osteosarcoma were later diagnosed (at the age of eight years) both within and outside the field of radiation. Examination of peripheral-leukocyte DNA revealed that the patient carried one mutated p53 allele with a mutation of serine to phenylalanine at codon 241. SSCP analysis of her family showed that the variant band caused by the mutant p53 allele was not present in either of her parents or her only sibling, suggesting that this mutation was also a new germline mutation (Fig. 2Figure 2
Pedigrees and SSCP Analyses of Germline Mutations of the p53 Gene in Two Patients with No Known Family History of Cancer.A).In the third patient, osteosarcoma was diagnosed at 18 years of age (Patient 3 in Table 1). His disease followed a rapid course, with multiple foci of osteosarcoma, and he could not be successfully treated. The variant band detected on SSCP analysis of his peripheral blood results from a transition from guanine to adenine at codon 245, causing the substitution of a serine residue where glycine normally occurs. The variant p53 allele was also found in his father and younger brother (Fig. 2B). The proband's father was healthy and in his mid-50s at the time of the study; although he had no history of cancer, he had numerous pigmented, benign nevi. The younger brother of the proband had a single osteosarcoma at the age of 18 years that was successfully treated; he also had skin lesions like those of his father. The proband would probably have been regarded as having sporadic osteosarcoma that was unremarkable until osteosarcoma also developed in his sibling.
Patients with an Unusual Personal or Family History of Cancer
We found five germline mutations of the p53 gene in patients who were known to have an unusual family history of cancer at the time of diagnosis. In the first patient, osteosarcoma was diagnosed at 19 years of age (Patient 4 in Table 1); he had a family history of cancer that was consistent with the Li—Fraumeni syndrome (Fig. 3Figure 3
Pedigree and Analysis of the Hereditary Germline Nonsense Mutation of the p53 Gene in Patient 4.A). The mutation he carried was an insertion of one cytosine in a stretch of five cytosines spanning codons 151 through 152 (Fig. 3C). This insertion leads to a frame-shift error and a premature stop codon downstream at codon 180. This change would presumably lead to the truncation of 212 amino acids from the p53 protein produced by the mutant allele. In addition to the proband, his apparently healthy 4-year-old daughter and 12-year-old nephew were also carriers of the same variant p53 allele (Fig. 3B); other affected family members could not be tested.Three additional nonsense mutations and one missense mutation were found in patients with a family history of cancer (Fig. 4Figure 4
Pedigrees and SSCP Analyses of Germline Mutations of the p53 Gene in Four Patients with a Family History of Cancer.). A deletion of two bases at codons 209 through 210 that results in a premature stop at codon 214 was found in a blood sample from an eight-year-old girl with malignant fibrous histiocytoma (Patient 5 in Table 1). On the paternal side of her family there was a history of cancer, especially tumors of neural-tissue origin, although cancer had never been diagnosed in the girl's father (Fig. 4A). In the second case, we identified an insertion of one cytosine in a stretch of six cytosines spanning codons 71 through 72 in a young woman with osteosarcoma (Patient 6 in Table 1 and Fig. 4B). This mutation results in a premature stop at codon 148. In the third case, a transversion from adenine to thymine at the first nucleotide in codon 120 was found in a patient with osteosarcoma in whom other primary tumors subsequently developed (Patient 7 in Table 1 and Fig. 4C). This mutation results directly in a new stop codon. Although other members of these three families were not available for DNA analysis, the mothers of both probands with osteosarcoma had premenopausal bilateral breast cancer (Fig. 4B and 4C).In the fourth case, a transition from cytosine to thymine at codon 282 was found on one p53 allele carried constitutionally (Patient 8 in Table 1). This mutation results in the substitution of a tryptophan where an arginine residue normally occurs in the p53 protein. Although this mutation is within the evolutionarily conserved regions of the p53 gene, it is downstream from the other missense mutations that we found and from all previously published germinal missense mutations in the p53 gene.3 , 4 , 23 The proband of this family (Patient 8) was given a diagnosis of osteosarcoma at the age of 10 years and had an extensive family history of malignant tumors with an unusual prevalence of gastric cancer on the paternal side. Our analysis showed that the germline mutation carried by the proband and her affected father was also present in her two apparently healthy sisters, who were 15 and 9 years old at the time of the study (Fig. 4D).
Polymorphisms in the Coding Sequences of the p53 Gene
In addition to the mutations described above that increase one's susceptibility to cancer, we found six hereditary variations in the coding sequences of the p53 gene (Table 2Table 2
Rare Polymorphisms in the Coding Sequence of the p53 Gene.*). One of them is the previously reported neutral polymorphism at codon 72 (72Pro to 72Arg), with no known or predicted phenotypic consequences.28 Two of the other five changes are silent mutations: one at codon 36 (36Pro to 36Pro) and a second at codon 213 (213Arg to 213Arg). We found three novel nucleotide substitutions that result in amino acid substitutions. Two lead to conservative amino acid changes: one at codon 11(1 lGlu to 11Gln), and the other at codon 31 (31Val to 31Ile). A third amino acid change at codon 49 (49Asp to 49His) is a relatively unconservative substitution. All three alleles were found in both the patients with sarcoma and the healthy control populations that we studied. This variant allele could have phenotypic consequences, but our data do not exclude the possibility that it is a neutral polymorphism, since it lies outside the conserved regions of the protein.Discussion
Mutations of the p53 tumor-suppressor gene are common in the progression of many cancers, in which both recessive "loss of function" and dominant "activating" mutations may occur. Although evidence suggests that its role lies in the later stages of malignant progression,29 the recent discovery of germline mutations of this gene in cancer-prone families with the Li—Fraumeni syndrome raises several important questions.3 , 4 The first concerns the prevalence of germline mutations of the p53 gene among patients with sarcoma who have no unusual family history of cancer. In this study, three such patients were identified. Each was affected by osteosarcoma, and each carried a mutated p53 allele as a constitutional defect. Because one of these patients (Patient 2) had a personal history of cancer before the diagnosis of osteosarcoma, only 2 of the 8 carriers of germline p53 mutations identified among 196 patients with sarcoma in our study might be regarded as having sporadic cases of osteosarcoma at presentation. Therefore, the fraction of patients with apparently sporadic sarcoma who carry a germline mutation of the p53 gene is likely to be very low, on the order of 1 to 2 percent. For comparison, 12 percent of patients with simplex retinoblastoma carry new germline mutations of the retinoblastoma gene, and three fourths of the hereditary cases involve new germline mutations.2 The paucity of new germline mutations at the p53 locus detected in our study could indicate that many missense mutations in the p53 gene are lethal during embryogenesis, and only a limited number of subtle changes are nonlethal. Recent evidence supports this argument. One study has shown that p53 proteins with a substitution at codon 248, similar to three of seven germline missense mutations described elsewhere, do not form a complex with the wildtype protein.30 Since mutations of the p53 gene that act dominantly appear to inactivate the wild-type protein through the formation of this complex,31 it seems likely that codon 248 mutations are nonlethal and are allowed in the germline because they are recessive.
The unexpected finding of germline nonsense mutations of the p53 gene sheds additional light on the role of this gene in human cancer. There were equal numbers of nonsense and missense mutations of the p53 gene in the germline of patients with sarcoma. The clinical phenotype, penetrance, and spectrum of cancers associated with the two types of mutation were indistinguishable, as can be seen by comparing the pedigree of Patient 1 with that of Patient 4. The identification of nonsense mutations in the germline of these cancer-prone patients reveals that, although some areas with a high likelihood of having somatic missense p53 mutations15 may also carry an increased likelihood of having germline missense mutations, nonsense mutations in the p53 gene may be equally likely to predispose patients to cancer. It appears that although nonlethal germline missense mutations will be found only in strictly defined regions of the p53 gene, the target for heritable nonsense mutations is much larger — an issue that is of importance for future studies involving large-scale screening of patients with cancer for germline mutations of the p53 gene. In addition, we identified several polymorphisms in coding regions of the p53 gene; accurate predictions of the increased likelihood of cancer conferred by variant p53 alleles may require that such apparently "neutral" genetic polymorphisms be analyzed in terms of the hypothesized functions of the p53 protein7 8 9 10 and its potential transforming activity when mutated.21
The average age at diagnosis of seven patients with osteosarcoma who carried germline mutations was 15 years (range, 8 to 19). This is not remarkably different from the age of the patients with osteosarcoma in this study who did not have a p53 gene defect (18 years; range, 4 to 52). Thus, the presence of a germline mutation of the p53 gene does not necessarily accelerate the appearance of osteosarcoma in carriers, although it increases their overall susceptibility to the disease. On the other hand, in three of seven patients with osteosarcoma who carried a germline mutation multiple foci of tumor developed after the primary focus was treated; although metastasis cannot be excluded in all cases, several are thought to have been treatment-induced tumors. By analogy with retinoblastoma, it is reasonable to assume that cells with one mutant allele of the p53 gene are potentially more vulnerable than normal cells to iatrogenic carcinogens, such as anticancer drugs or ionizing radiation. In particular, ionizing radiation is a known cause of second, treatment-induced cancers in patients with retinoblastoma.32 This point may be relevant in the choice of treatments of patients with a germline mutation of the p53 gene, since the p53 gene is analogous to the retinoblastoma gene in its role as a tumor suppressor, and a recessive p53 mutation carried in the germline may be equally susceptible to malignant expression in somatic cells after radiation therapy.
The data shown for the family of Patient 3 emphasize the involvement of other environmental or genetic factors in the progression of sarcoma. In this family, both affected siblings inherited the same germline mutation from their father and had osteosarcoma at the same age, but the course of disease was remarkably different in each. Furthermore, the father in this family was a carrier and was apparently healthy in midlife, indicating that there may be considerable clinical heterogeneity within a single family. Familial cancer associated with the inheritance of a variant p53 allele may be heterogeneous, and it could be subject to environmental factors such as those that drive nonfamilial cancer. In one family (that of Patient 8), for example, there was a preponderance of gastric cancer, which is thought to be rare in families affected by the Li—Fraumeni syndrome. This family is from Japan, where gastric cancer is much more common than in North America33; recently, somatic mutations of the p53 gene have been reported in advanced-stage gastric cancers.34 , 35 It is tempting to speculate that different genetic or environmental factors determine the types of tumors that will occur in patients who carry the same p53 gene mutation.
Among 181 randomly selected patients with sarcoma, we identified only 2 (Patients 6 and 8) who had germline mutations of the p53 gene. This fraction (1 to 2 percent) is small but not negligible; both patients had a family history of cancer. Six of the 15 patients with sarcoma that we identified as potentially prone to cancer, on the basis of a personal or family history of cancer, were carriers of a mutated p53 allele. This fraction should not be taken as representative of any specific group of patients or of cancer-prone patients in general, since our ascertainment of patients who are potentially prone to cancer was not based on a specific clinical definition. We found germline mutations of the p53 gene both in families that fit the definition of the Li—Fraumeni syndrome and in families that did not; thus, the clinical picture appears to be broader than was previously suspected.
Knowledge of the germline mutation carried by these patients may allow accurate DNA-based carrier diagnosis and genetic counseling in their families. However, the occurrence of unaffected carriers of cancer-predisposing mutations in these families raises serious questions about what represents appropriate methods of cancer surveillance and counseling for these persons. We believe that the screening method we used, which was based on a combination of SSCP and direct genomic sequencing, is rapid and sensitive enough to be used for large-scale screening and may help address some of the questions this study has raised.
Supported by grants from the National Institutes of Health (D.W.Y., R.R.W., J.B.L.), the American Cancer Society (D.W.Y.), and the Japanese Ministry of Education, Science, and Culture (M.S.S., T. Yamamuro). Dr. Yandell is a Research to Prevent Blindness Dolly Green scholar.
We are indebted to Drs. Y. Kotoura, N. Takada, N. Kawaguchi, Y. Kaneko, B. Ritchie, D. Brachman, I. Spiro, A. Rosenberg, B. Leventhal, and H. Halprin for their assistance in collecting clinical materials; to Drs. T. Dryja, B. Ludeke, R. Chung, J. Whaley, and G. Cowley for their cooperation and technical advice; to Drs. L. Crawford and S. Tuck for genomic clones containing p53; and to Dr. L.C. Strong for helpful discussions regarding the manuscript.
Source Information
From the Massachusetts Eye and Ear Infirmary, Boston (J.T., S.H.D., R.L.B., G.E.H., D.W.Y.); the Radiation Biology Center (T. Yamaguchi, K.I., M.S.S.) and Faculty of Medicine (T. Yamaguchi, T. Yamamuro), Kyoto University, Kyoto, Japan; Memorial Sloan-Kettering Cancer Center, New York (P.A.M.); Harvard School of Public Health, Boston (J.B.L., D.W.Y.); and the University of Chicago Medical Center, Chicago (R.R.W.). Address reprint requests to Dr. Yandell at the Howe Laboratory, Massachusetts Eye and Ear Infirmary, 243 Charles St., Boston, MA 02114.
- References
References
1
Knudson
Web of Science | Medline2
Vogel
CrossRef | Web of Science | Medline3
Matkin
CrossRef | Web of Science4
Srivastava
CrossRef | Web of Science | Medline5
Li
Web of Science | Medline6
Lane
CrossRef | Web of Science | Medline7
Fields
CrossRef | Web of Science | Medline8
Raycroft
CrossRef | Web of Science | Medline9
Iguchi-Ariga
Web of Science | Medline10
Kern
CrossRef | Web of Science | Medline11
Finlay
CrossRef | Web of Science | Medline12
Baker
CrossRef | Web of Science | Medline13
Masuda
CrossRef | Web of Science | Medline14
Nigro
CrossRef | Web of Science | Medline15
Ahuja
CrossRef | Web of Science | Medline16
Mulligan
CrossRef | Web of Science | Medline17
Tamboli
Web of Science | Medline18
Goorin
Full Text | Web of Science | Medline19
Crist
Web of Science | Medline20
Mowat
CrossRef | Web of Science | Medline21
Hinds
Web of Science | Medline22
Soussi
Web of Science | Medline23
Metzger
CrossRef | Web of Science | Medline24
Kunkel
CrossRef | Web of Science | Medline25
Saiki
CrossRef | Web of Science | Medline26
Orita
CrossRef | Web of Science | Medline27
Yandell
Web of Science | Medline28
Matlashewski
Web of Science | Medline29
Baker
Web of Science | Medline30
Milner
CrossRef | Web of Science | Medline31
Kraiss
Web of Science | Medline32
Abramson
Web of Science | Medline33
Takebe H, Utsunomiya J, eds. Genetics of human tumors in Japan. Gann monograph on cancer research no. 35. Tokyo: Japan Scientific Societies Press, 1988.
34
Tamura
Web of Science | Medline35
Kim
CrossRef | Web of Science
- Citing Articles (79)
Citing Articles
1
Christopher Poremba. 2011. , 353-370.
CrossRef2
François Goldwasser. 2011. Clinical Approach in Soft Tissue Tumors. , 1-19.
CrossRef3
L. M. Almquist, M. R. Karagas, B. C. Christensen, M. M. Welsh, A. E. Perry, C. A. Storm, H. H. Nelson. (2011) The role of TP53 and MDM2 polymorphisms in TP53 mutagenesis and risk of non-melanoma skin cancer. Carcinogenesis 32:3, 327-330
CrossRef4
Anne E. Newcomer, Diane Dylinski, Brian P. Rubin, Michael J. Joyce, Gerald Hoeltge, Boris Bershadsky, Steven A. Lietman. (2011) Prognosticators in thigh soft tissue sarcomas. Journal of Surgical Oncology 103:1, 85-91
CrossRef5
Simona Ognjanovic, Magali Oliver, Tracy L. Bergemann, Pierre Hainaut. (2011) Sarcomas in TP53 germline mutation carriers. Cancern/a-n/a
CrossRef6
Mandy Geryk-Hall, Dennis P. M. Hughes. (2009) Critical signaling pathways in bone sarcoma: Candidates for therapeutic interventions. Current Oncology Reports 11:6, 446-453
CrossRef7
Emilios E. Pakos, Andreas D. Nearchou, Robert J. Grimer, Haris D. Koumoullis, Adesegun Abudu, Jos A.M. Bramer, Lee M. Jeys, Alessandro Franchi, Guido Scoccianti, Domenico Campanacci, Rodolfo Capanna, Jorge Aparicio, Marie-Dominique Tabone, Gerold Holzer, Fashid Abdolvahab, Philipp Funovics, Martin Dominkus, Inci Ilhan, Su G. Berrak, Ana Patino-Garcia, Luis Sierrasesumaga, Mikel San-Julian, Moira Garraus, Antonio Sergio Petrilli, Reynaldo Jesus Garcia Filho, Carla Renata Pacheco Donato Macedo, Maria Teresa de Seixas Alves, Sven Seiwerth, Rajaram Nagarajan, Timothy P. Cripe, John P.A. Ioannidis. (2009) Prognostic factors and outcomes for osteosarcoma: An international collaboration. European Journal of Cancer 45:13, 2367-2375
CrossRef8
Mwe Mwe Chao, John E. Levine, Robert E. Ruiz, Wendy K. Kohlmann, Matthew A. Bower, Elizabeth M. Petty, Rajen J. Mody. (2007) Malignant triton tumor in a patient with Li-Fraumeni syndrome and a novelTP53 mutation. Pediatric Blood & Cancer 49:7, 1000-1004
CrossRef9
Ikuo Kudawara, Akihiko Matsumine, Kenji Ohzono. (2007) Analysis of germline and tumor mutations of p53 gene in familial occurrence of soft tissue sarcomas. Journal of Surgical Oncology 95:4, 347-350
CrossRef10
Karl T. Kelsey, Tomoko Hirao, Shuya Hirao, Tara Devi-Ashok, Heather H. Nelson, Angeline Andrew, Joanne Colt, Dalsu Baris, J. Steven Morris, Alan Schned, Margaret Karagas. (2005) TP53 alterations and patterns of carcinogen exposure in a U.S. population-based study of bladder cancer. International Journal of Cancer 117:3, 370-375
CrossRef11
Purificación García-Miguel, y Manuel López Santamaría. (2005) Estado actual del diagnóstico y tratamiento del hepatoblastoma. Clinical and Translational Oncology 7:7, 328-334
CrossRef12
Koichi Nishijo, Tomitaka Nakayama, Tomoki Aoyama, Takeshi Okamoto, Tatsuya Ishibe, Ko Yasura, Yasuko Shima, Kotaro R. Shibata, Tadao Tsuboyama, Takashi Nakamura, Junya Toguchida. (2004) Mutation analysis of theRECQL4 gene in sporadic osteosarcomas. International Journal of Cancer 111:3, 367-372
CrossRef13
Ana Patiño-García, Elena Sotillo Piñeiro, Marta Zalacaín Díez, Leire Gárate Iturriagagoitia, Federico Antillón Klüssmann, Luis Sierrasesúmaga Ariznabarreta. (2003) Genetic and Epigenetic Alterations of the Cell Cycle Regulators and Tumor Suppressor Genes in Pediatric Osteosarcomas. Journal of Pediatric Hematology/Oncology 25:5, 362-367
CrossRef14
Shih-Jen Hwang, Guillermina Lozano, Christopher I. Amos, Louise C. Strong. (2003) Germline p53 Mutations in a Cohort with Childhood Sarcoma: Sex Differences in Cancer Risk. The American Journal of Human Genetics 72:4, 975-983
CrossRef15
Anna Patrikidou, Jon Bennett, Patrick Abou-Sleiman, Joy D.A. Delhanty, Malcolm Harris. (2002) A novel, de novo germline TP53 mutation in a rare presentation of the Li-Fraumeni syndrome in the maxilla. Oral Oncology 38:4, 383-390
CrossRef16
Brian D Ragland, Walter C Bell, Robert R Lopez, Gene P Siegal. (2002) Cytogenetics and Molecular Biology of Osteosarcoma. Laboratory Investigation 82:4, 365-373
CrossRef17
Maria do Socorro Maciel, Leda C. Viegas, Suely Nonogaki, Inês N. Nishimoto, Fauzer S. Abrão, Mário Mourão Neto, M. Mitzi Brentani. (2002) p53 expression is a factor for prognostic assessment in breast sarcoma. Breast Cancer Research and Treatment 71:3, 193-202
CrossRef18
Kenji Kimura, Kazuya Shinmura, Kimio Yoshimura, Kimihiro Shimizu, Hitoshi Katai, Yasuo Beppu, Hideshige Moriya, Jun Yokota. (2000) Absence of Germline CHK2 Mutations in Familial Gastric Cancer. Cancer Science 91:9, 875-879
CrossRef19
Sevan Hopyan, Nalan Gokgoz, Robert Stuart Bell, Irene Louise Andrulis, Benjamin Aaron Alman, Jay Steven Wunder. (1999) Expression of osteocalcin and its transcriptional regulatorscore-binding factor alpha 1 andMSX2 in osteoid-forming tumours. Journal of Orthopaedic Research 17:5, 633-638
CrossRef20
Andreas K. Nussler, Gianna Vergani, Susanne M. Gollin, Kenneth Dorko, Susanne Gansauge, Sidney M. Morris, Antony J. Demetris, Minoru Nomoto, Hans G. Beger, Stephen C. Strom. (1999) Isolation and characterization of a human hepatic epithelial-like cell line (AKN-1) from a normal liver. In Vitro Cellular & Developmental Biology - Animal 35:4, 190-197
CrossRef21
S.J. Ham, W.T.A. van der Graaf, E. Pras, W.M. Molenaar, E. van den Berg, H.J. Hoekstra. (1998) Soft tissue sarcoma of the extremities. A multimodality diagnostic and therapeutic approach. Cancer Treatment Reviews 24:6, 373-391
CrossRef22
S.J. Ham, H. Schraffordt Koops, W.T.A. van der Graaf, J.R. van Horn, L. Postma, H.J. Hoekstra. (1998) Historical, current and future aspects of osteosarcoma treatment. European Journal of Surgical Oncology (EJSO) 24:6, 584-600
CrossRef23
George Kijima, Yoshinori Murakami, Noriaki Ohuchi, Susumu Satomi, Takao Sekiya. (1998) Nonsense Mutation at Codon 63 of the BRCA1 Gene in Japanese Breast Cancer Patients. Cancer Science 89:8, 837-841
CrossRef24
Kathleen J. Smith, Terry L. Barrett, William F. Smith, Henry M. Skelton. (1998) A Review of Tumor Suppressor Genes in Cutaneous Neoplasms With Emphasis on Cell Cycle Regulators. The American Journal of Dermatopathology 20:3, 302-313
CrossRef25
Stephanie S. Martin, W. Grear Hurt, Lora K. Hedges, Merlin G. Butler, Herbert S. Schwartz. (1998) Microsatellite instability in sarcomas. Annals of Surgical Oncology 5:4, 356-360
CrossRef26
MAKOTO AKASHI, H. PHILLIP KOEFFLER. (1998) Li-Fraumeni Syndrome and the Role of the p53 Tumor Suppressor Gene in Cancer Susceptibility. Clinical Obstetrics and Gynecology 41:1, 172-199
CrossRef27
Kim E. Nichols, Frederick P. Li, Daniel A. Haber, Lisa Diller. (1998) Childhoood cancer predisposition: Applications of molecular testing and future implications. The Journal of Pediatrics 132:3, 389-397
CrossRef28
Ryohei Yokoyama, Regine Schneider-Stock, Kathrin Radig, Thomas Wex, Albert Roessner. (1998) Clinicopathologiclmplications of MDM2, p53 and K-ras gene alterations in osteosarcomas: MDM2 amplification and p53 mutations found in progressive tumors. Pathology - Research and Practice 194:9, 615-621
CrossRef29
Dolphine Oda, Linda M. Bavisotto, Rodney A. Schmidt, Michael McNutt, James D. Bruckner, Ernest U. Conrad, Ernest A. Weymuller. (1997) Head and neck osteosarcoma at the University of Washington. Head & Neck 19:6, 513-523
CrossRef30
Reinaldo Manhani, Lilian Maria Cristofani, Vicente Filho, Israel Bendit. (1997) Concomitant p53 mutation and MYCN amplification in neuroblastoma. Medical and Pediatric Oncology 29:3, 206-207
CrossRef31
C. S. Brock, M. Bower. (1997) Current perspectives in gliomas. Medical Oncology 14:2, 103-120
CrossRef32
RS Cornelis, M van Vliet, MJ van de Vijver, HFA Vasen, PA Voute, B Top, P. Meera Khan, P Devilee, CJ Cornelisse. (1997) Three germline mutations in theTP53 gene. Human Mutation 9:2, 157-163
CrossRef33
Paul L. Baron, Christopher E. Gates, Carolyn E. Reed, Roberta L. D. Dikeman, Jay J. Drosieko, Roger N. Passmore, Jonathan S. Bromberg, Mark C. Willingham. (1997) p53 Overexpression in squamous cell carcinoma of the esophagus. Annals of Surgical Oncology 4:1, 37-45
CrossRef34
Eleftherios P Diamandis. (1997) Clinical applications of tumor suppressor genes and oncogenes in cancer. Clinica Chimica Acta 257:2, 157-180
CrossRef35
Carl W. Miller, Abdulkarim Aslo, Alvina Won, Michael Tan, Beatrice Lampkin, H. Phillip Koefflar. (1996) Alterations of thep53, Rb andMDM2 genes in osteosarcoms. Journal of Cancer Research and Clinical Oncology 122:9, 559-565
CrossRef36
S Moore. (1996) Absence of germline mutations in exons 5–9 of the p53 gene in patients with Li-Fraumeni-like (SBLA) and familial adenomatous polyposis heritable cancer syndromes. Cancer Genetics and Cytogenetics 90:2, 125-129
CrossRef37
Concha López-Ginés, Carmen Carda-Batalla, Lola López-Terrada, Antonio Llombart-Bosch. (1996) Presence of double minutes and monosomy 17p in xenografted human osteosarcomas. Cancer Genetics and Cytogenetics 90:1, 57-62
CrossRef38
D Zhu. (1996) No evidence for the amplifications of MDM2 and C-myc genes involved in the genetic susceptibility to esophageal cancer in a high-risk area of North China. Cancer Genetics and Cytogenetics 89:2, 184-185
CrossRef39
(1996) Evidence of a hereditaryp53 syndrome in cancer-prone families. International Journal of Cancer 65:4, 554-557
CrossRef40
Atsushi Sasaki, Makoto Nagashima, Masayuki Shiseki, Hitoshi Katai, Keiichi Maruyama, Ritsuko Iwanaga, Yoshimitsu Akiyama, Yasuhito Yuasa, Jun Yokota. (1996) Microsatellite instability in gastric cancer prone families. Cancer Letters 99:2, 169-175
CrossRef41
C.J. Cornelisse, R.S. Cornelis, P. Devilee. (1996) Genes Responsible for Familial Breast Cancer. Pathology - Research and Practice 192:7, 684-693
CrossRef42
David W. Yandell, Christopher Poremba. (1996) Genetics of retinoblastoma: Implications for other human cancers. Medical and Pediatric Oncology 27:S1, 25-28
CrossRef43
You-Jun Li, Marc Sanson, Khê Hoang-Xaun, Jean-Yves Delattre, Michel Poisson, Gilles Thomas, Richard Hamelin. (1995) Incidence of germ-linep53 mutations in patients with gliomas. International Journal of Cancer 64:6, 383-387
CrossRef44
SHOSHANA PELLER, YULIA KOPILOVA, SAM SLUTZKI, ARIEL HALEVY, KATIA KVITKO, VARDA ROTTER. (1995) A Novel Polymorphism in Intron 6 of the Human p53 Gene: A Possible Association with Cancer Predisposition and Susceptibility. DNA and Cell Biology 14:12, 983-990
CrossRef45
Cornelius Schmaltz, Siegfried Sauter, Oliver Opitz, Dieter Harms, Bernhard Kremens, Michael Lohner, Klaus Metz, Matthias Brandis, Charlotte Niemeyer. (1995) Pleuro-pulmonary blastoma: A case report and review of the literature. Medical and Pediatric Oncology 25:6, 479-484
CrossRef46
Wolfram G. Scheurlen, Jürgen Krauss, Joachim Kühl. (1995) No preferential loss of one parental allele of chromosome 17p13.3 in childhood medulloblastoma. International Journal of Cancer 63:3, 372-374
CrossRef47
C. Poremba, D. W. Yandell, W. Mellin, K. W. Schmid, B. Reers, W. Böcker, B. Dockhorn-Dworniczak. (1995) Adenocarcinoma of the Cardia in a Young Man: Detection of Somatic p53 Mutation by Immunohistochemistry and Automated Direct Sequencing. Pathology - Research and Practice 191:10, 1004-1009
CrossRef48
P Chen. (1995) Constitutional p53 mutations associated with brain tumors in young adults. Cancer Genetics and Cytogenetics 82:2, 106-115
CrossRef49
Eleftherios P. Diamandis. (1995) Clinical applications of the p53 tumor suppressor gene. Clinica Chimica Acta 237:1-2, 79-90
CrossRef50
Benedikt L. Ziegler, Melanie Weiss, Stefan Thoma, Christa Lamping, Theodor M. Fliedner. (1995) Chapter 8: Perspectives for the use of biological indicators for the assessment of radiation induced responses and impairments: Biologic indicators of exposure: Are markers associated with oncogenesis useful as biologic markers of effect?. Stem Cells 13:S1, 326-338
CrossRef51
Toyoaki Uchida, Chieki Wada, Hironori Ishida, Chunxi Wang, Shin Egawa, Eiji Yokoyama, Toru Kameya, Ken Koshiba. (1995) p53 Mutations and Prognosis in Bladder Tumors. The Journal of Urology 153:4, 1097-1104
CrossRef52
Christine Le Bihan, Céline Moutou, Laurence Brugières, Jean Feunteun, Catherine Bonaïti-Pellié. (1995) ARCAD: A method for estimating age-dependent disease risk associated with mutation carrier status from family data. Genetic Epidemiology 12:1, 13-25
CrossRef53
Inci Ilhan, Selim Erekul, Sabri Ateşalp, Osman Ilhan, Nejat Akar. (1995) p53 Codon 213 (A-G) Polymorphism in a Turkish Population. Pediatric Hematology-Oncology 12:5, 499-501
CrossRef54
Vijay S. Tonk, Kathleen S. Wilson, Charles F. Timmons, Nancy R. Schneider. (1994) Trisomy 2, trisomy 20, and del(17p) as sole chromosomal abnormalities in three cases of hepatoblastoma. Genes, Chromosomes and Cancer 11:3, 199-202
CrossRef55
Raphael E. Pollock. (1994) Molecular determinants of soft tissue sarcoma proliferation. Seminars in Surgical Oncology 10:5, 315-322
CrossRef56
Richard Hamelin, Fernande Barichard, Isabelle Henry, Claudine Junien, Gilles Thomas. (1994) Single base pair germ-line deletion in the p53 gene in a cancer predisposed family. Human Genetics 94:1, 88-90
CrossRef57
Massimo Loda. (1994) Polymerase chain reaction-based methods for the detection of mutations in oncogenes and tumor suppressor genes. Human Pathology 25:6, 564-571
CrossRef58
Alan G. Casson, Nancy Kerkvliet, Frances O'Malley, Richard Inculet, Michael Troster. (1994) P53 immunoreactivity in carcinosarcoma of the esophagus. Journal of Surgical Oncology 56:2, 132-135
CrossRef59
Hitoshi Nakagawa, Hiroshi Fujii, Hiroyuki Nakai, Johji Inazawa, Shinichi Misawa. (1994) Myelomonocytic crisis with t(5;17) and a p53 mutation in a patient with chronic myelogenous leukemia. American Journal of Hematology 45:4, 335-340
CrossRef60
W. Alexandra Hoogerwerf, Anita L. Hawkins, Constance A. Griffin, Elizabeth J. Perlman. (1994) Chromosome analysis of nine osteosarcomas. Genes, Chromosomes and Cancer 9:2, 88-92
CrossRef61
Walter W. Piegorsch, Clarice R. Weinberg, Jack A. Taylor. (1994) Non-hierarchical logistic models and case-only designs for assessing susceptibility in population-based case-control studies. Statistics in Medicine 13:2, 153-162
CrossRef62
Carolyn L. Russo, James McIntyre, Allen M. Goorin, Michael P. Link, Mark C. Gebhardt, Stephen H. Friend. (1994) Secondary breast cancer in patients presenting with osteosarcoma: Possible involvement of germline p53 mutations. Medical and Pediatric Oncology 23:4, 354-358
CrossRef63
Alejandro Espaillat, Soon Jin Lee, Victor Arrunategui-Correa, C. Stephen Foster, Albert Vitale, Sabatino Dimeo, Robert B. Colvin. (1994) Expression of fibronectin isoforms in rat cornea after an epithelial-scrape wound. Current Eye Research 13:5, 325-330
CrossRef64
Michele Harvey, Mark J. McArthur, Charles A. Montgomery, Janet S. Butel, Allan Bradley, Lawrence A. Donehower. (1993) Spontaneous and carcinogen–induced tumorigenesis in p53–deficient mice. Nature Genetics 5:3, 225-229
CrossRef65
Harris, Curtis C.Hollstein, Monica. (1993) Clinical Implications of the p53 Tumor-Suppressor Gene. New England Journal of Medicine 329:18, 1318-1327
Full Text66
Chikashi Ishioka, Thierry Frebourg, Yu-Xin Yan, Marc Vidal, Stephen H. Friend, Susanne Schmidt, Richard Iggo. (1993) Screening patients for heterozygous p53 mutations using a functional assay in yeast. Nature Genetics 5:2, 124-129
CrossRef67
Masayuki Shiseki, Ryo Nishikawa, Hiroshi Yamamoto, Atsushi Ochiai, Haruhiko Sugimura, Nobuyuki Shitara, Yuichi Sameshima, Hideaki Mizoguchi, Takashi Sugimura, Jun Yokota. (1993) Germ-line p53 mutation is uncommon in patients with triple primary cancers. Cancer Letters 73:1, 51-57
CrossRef68
Lawrence A. Donehower, Allan Bradley. (1993) The tumore suppressor p53. Biochimica et Biophysica Acta (BBA) - Reviews on Cancer 1155:2, 181-205
CrossRef69
Ann L. Hartley, Jillian M. Birch, Val Blair, Anna M. Kelsey, Martin Harris, Patricia H. Morris Jones. (1993) Patterns of cancer in the families of children with soft tissue sarcoma. Cancer 72:3, 923-930
CrossRef70
Spiro, Ira J.Yandell, David W.Li, ChuanSaini, SanjayFerry, JudithPowelson, JohnKatkov, William N.Cosimi, A. Benedict. (1993) Lymphoma of Donor Origin Occurring in the Porta Hepatis of a Transplanted Liver. New England Journal of Medicine 329:1, 27-29
Full Text71
Jochen Schütte, Bertram Opalka, Reinhard Becher, Walter Bardenheuer, Susanne Szymanski, Andreas Lux, Siegfried Seeber. (1993) Analysis of the p53 gene in patients with isochromosome 17q and Ph1-positive or -negative myeloid leukemia. Leukemia Research 17:6, 533-539
CrossRef72
Ju Chen, Amrik Sahota, Glenn F. Martin, Masayuki Hakoda, Naoyuki Kamatani, Peter J. Stambrook, Jay A. Tischfield. (1993) Analysis of germline and in vivo somatic mutations in the human adenine phosphoribosyltransferase gene: Mutational hot spots at the intron 4 splice donor site and at codon 87. Mutation Research/Fundamental and Molecular Mechanisms of Mutagenesis 287:2, 217-225
CrossRef73
Sang Wook Choi, Prodromos Hytiroglou, Stephen A. Geller, Sun Moo Kim, Kyu Won Chung, Doo Ho Park, Neil D. Theise, Swan N. Thung. (1993) The expression of p53 antigen in primary malignant epithelial tumors of the liver: an immunohistochemical study. Liver 13:3, 172-176
CrossRef74
(1993) REVIEW. Biological Chemistry Hoppe-Seyler 374:1-6, 227-236
CrossRef75
Birgitte Smith-Sørensen, Mark C. Gebhardt, Peter Kloen, Jim McIntyre, Fernando Aguilar, Peter Cerutti, Anne-Lise Børresen. (1993) Screening for TP53 mutations in osteosarcomas using constant denaturant gel electrophoresis (CDGE). Human Mutation 2:4, 274-285
CrossRef76
Ikuo Yana, Tsutomu Nakamura, Eisei Shin, Katsu Karakawa, Hiroki Kurahashi, Yoshihiro Kurita, Tetsuro Kobayashi, Takesada Mori, Isamu Nishisho, Shin-ichiro Takai. (1992) Inactivation of the p53 Gene Is not Required for Tumorigenesis of Medullary Thyroid Carcinoma or Pheochromocytoma. Cancer Science 83:11, 1113-1116
CrossRef77
Orna Mor, Zehava Grossman, Orit Jakobovitz, Frida Brok-Simoni, Gideon Rechavi. (1992) Artifactual p53 point mutations: possible effect of gene secondary structure on PCR and direct sequence analysis. The Lancet 340:8829, 1236
CrossRef78
Carole Hennessy Martz. (1992) Introduction. Seminars in Oncology Nursing 8:4, 227-228
CrossRef79
Levine, Arnold J.. (1992) The p53 Tumor-Suppressor Gene. New England Journal of Medicine 326:20, 1350-1352
Full Text
