TP53 mutations and loss of heterozygosity of chromosome 17 in colorectal tumors

Yamamoto, Lidia; Lopes, Ademar A.; Harb-Gama, Angelito; Nagai, Maria A.

doi:10.1590/S0100-84551996000400022

Abstract

The incidence of TP53 point mutations and loss of heterozygosity (LOH) of chromosome 17 in colorectal tumors was determined in a group of Brazilian patients. We screened DNA samples from tumors and distal normal mucosa of 39 patients with colorectal cancer, for TP53 mutations by PCR-SSCP (single-strand conformation polymorphism) analysis. Chromosome 17 LOH was investigated using six PCR-based polymorphic markers and one VNTR probe. TP53 mutations were demonstrated in 15/39 of the cases. Mutations were distributed among all exons examined (five to eight), the majority of them being G/C to A/T transitions. LOH of chromosome 17p and 17q was detected in 70 and 46% of the tumors, respectively. There was a significant association between TP53 mutations and LOH in chromosome 17p (P = 0.0035) and 17q (P = 0.03). Although no correlation was observed between TP53 genetic alterations and clinical/ pathological characteristics, the association of TP53 mutations with loss of both chromosome 17 arms may indicate that TP53 inactivation provokes an unstable phenotype in tumor cells in colorectal tumors.

TP53 mutations; heterozygosity; chromosome 17; colorectal tumors

Full text in pdf format

TP53 mutations and loss of heterozygosity of chromosome 17 in colorectal tumors

Lidia Yamamoto^I; Ademar A. Lopes ^II; Angelito Harb-Gama^III; Maria A. Nagai^I

^IDisciplina de Oncologia, Departamento de Radiologia, Faculdade de Medicina, Universidade de São Paulo, Av. Dr. Arnaldo 455, 40 andar, 01246-093 São Paulo, SP, Brasil. Fax: (55) (11) 282-6580. Send correspondence to M.A.N.

^IIFundação Antônio Prudente, São Paulo, SP, Brasil

^IIIDepartamento de Gastroenterologia, Faculdade de Medicina, Universidade de São Paulo, São Paulo, SP, Brasil

ABSTRACT

The incidence of TP53 point mutations and loss of heterozygosity (LOH) of chromosome 17 in colorectal tumors was determined in a group of Brazilian patients. We screened DNA samples from tumors and distal normal mucosa of 39 patients with colorectal cancer, for TP53 mutations by PCR-SSCP (single-strand conformation polymorphism) analysis. Chromosome 17 LOH was investigated using six PCR-based polymorphic markers and one VNTR probe. TP53 mutations were demonstrated in 15/39 of the cases. Mutations were distributed among all exons examined (five to eight), the majority of them being G/C to A/T transitions. LOH of chromosome 17p and 17q was detected in 70 and 46% of the tumors, respectively. There was a significant association between TP53 mutations and LOH in chromosome 17p (P = 0.0035) and 17q (P = 0.03). Although no correlation was observed between TP53 genetic alterations and clinical/ pathological characteristics, the association of TP53 mutations with loss of both chromosome 17 arms may indicate that TP53 inactivation provokes an unstable phenotype in tumor cells in colorectal tumors.

Keywords: TP53 mutations; heterozygosity; chromosome 17; colorectal tumors.

Baker, S.J., Preisinger, A.C., Jessup, J.M., Paraskeva, C., Markowitz, S., Wilson, J.K.V., Hamilton, S. and Vogelstein, B. (1990). p53 gene mutations occur in combination with 17p allelic deletions as late events in colorectal tumorigenesis. Cancer Res. 50: 7717-7722.
Bartek, J., Bartkova, J., Vojtesek, B., Stakova, Z., Lukas, J., Rejthar, A., Kovarik, J., Midgley, C.A., Gannon, J.V. and Lane, D.P. (1991). Aberrant expression of the p53 oncoprotein is a common feature of a wide spectrum of human malignancies. Oncogene 6: 1699-1703.
Bedi, A., Pasricha, P.J., Akhtar, A.J., Barber, J.P., Bedi, G.C., Giardiello, F.M., Zehnbauer, S.R. and Jones, R.J. (1995). Inhibition of apoptosis during development of colorectal cancer. Cancer Res. 55: 1811-1816.
Bosari, S., Viale, G., Bossi, P., Maggioni, M., Coggi, G., Murray, J.J. and Lee, A.K.C. (1994). Cytoplasmic accumulation of p53 protein: an independent prognostic indicator in colorectal adenomas. J. Natl. Cancer Inst. 86: 681-687.
Cho, Y., Gorina, S., Jeffrey, P.D. and Pavletich, N.P. (1994). Crystal structure of a p53 tumor suppressor-DNA complex: Understanding tumorigenic mutations. Science 265: 346-355.
Cohn, K.H., Wang, F., DeSoto-Lapaix, F., Solomon, W.B., Patterson, L.G., Arnold, M.R., Wemar, J., Feldman, J.G., Levy, A.T., Leone, A. and Steeg, P.S. (1991). Association of nm23-H1 allelic deletions with distant metastases in colorectal carcinoma. Lancet 338: 722-724.
Coulondre, C., Miller, J.H., Farabaugh, P.J. and Gilbert, W. (1978). Molecular basis of base substitution hotspots in Escherichia coli. Nature 274: 775-780.
Cunningham, J., Lust, J.A., Schaid, D.J., Bren, G.D., Carpenter, H.A., Rizza, E., Kovach, J.S. and Thibodeau, S.N. (1992). Expression of p53 and 17p allelic loss in colorectal carcinoma. Cancer Res. 52: 1974-1980.
Eyfjord, J.E., Thorlacius, S., Steinarsdottir, M., Valgardsdottir, R., Ogmundsdottir, H.M. and Anamthawat-Jonsson, K. (1995). p53 abnormalities and genomic instability in primary human breast carcinomas. Cancer Res. 55: 646-651.
Fearon, E.R. and Vogelstein, B. (1990). A genetic model for colorectal tumorigenesis. Cell 61: 759-767.
Finlay, C.A., Hinds, P.W., Tan, T.-H., Eliyahu, D., Oren, D. and Levine, A.J. (1988). Activating mutations for transformation by p53 produce a gene product that forms an hsc70-p53 complex with an altered half-life. Mol. Cell. Biol. 8: 531-539.
Futreal, P.A., Barret, J.C. and Weiseman, R.W. (1991). An Alu polymorphism intragenic to the TP53 gene. Nucleic Acids Res. 19: 6977.
Gannon, J.V., Greaves, R., Iggo, R. and Lane, D.P. (1990). Activating mutations in p53 produce a common conformational effect. A monoclonal antibody specific for the mutant form. EMBO J. 9: 1595-1602.
Goh, H.-S., Chan, C.-S., Khine, K. and Smith, D.R. (1994). p53 and behaviour of colorectal cancer. Lancet 344: 233-234.
Hall, J.M., Friedman, L., Guenther, C., Lee, M.K., Weber, J.L., Black, D.M. and King, M.-C. (1992). Closing in on a breast cancer gene on chromosome 17q. Am. J. Hum. Genet. 50: 1235-1242.
Harris, C.C. and Hollstein, M. (1993). Clinical implications of the p53 tumor suppressor gene. N. Engl. J. Med. 329: 1318-1327.
Hasegawa, H., Ueda, M., Furukawa, K., Watanabe, M., Teramoto, T., Mukai, M. and Kitajima, M. (1995). p53 gene mutations in early colorectal carcinoma. De novo vs. adenoma-carcinoma sequence. Int. J. Cancer 64: 47-51.
Hoffman, B. and Liebermann, D.A. (1994). Molecular controls of apoptosis: differentiation/growth arrest primary response genes, proto-oncogenes, and tumor suppressor genes as positive and negative modulators. Oncogene 9: 1807-1812.
Hollstein, M., Sidransky, D., Vogelstein, B. and Harris, C.C. (1991). p53 in human cancers. Science 253: 49-53.
Jones, M.H. and Nakamura, Y. (1992). Detection of loss of heterozygosity at the human TP53 locus using a dinucleotide repeat. Genes Chromosom. Cancer 5: 89-90.
Khine, K., Smith, D.R. and Goh, H.-S. (1993). High frequency of allelic deletion on chromosome 17p in advanced colorectal cancer. Cancer 73: 28-35.
Kikuchi-Yanoshita, R., Konishi, M., Ito, S., Seki, M., Tanaka, K., Maeda, Y., Iino, H., Fukayama, M., Koike, M., Mori, T., Sakamura, H., Fukunari, H., Iwama, T. and Miyaki, M. (1992). Genetic changes of both p53 alleles associated with the conversion from colorectal adenoma to early carcinoma in familial adenomatous polyposis and non-familial adenomatous polyposis patients. Cancer Res. 52: 3965-3967.
Knudson, A.G. (1971). Mutation and cancer: Statistical study of retinoblastoma. Proc. Natl. Acad. Sci. USA 68: 820-823.
Lane, D.P. (1994). The regulation of p53 function: Steiner award lecture. Int. J. Cancer 57: 623-627.
Levine, A.J., Perry, M.P., Chang, A., Silver, A., Dittmer, D., Wu, M. and Welsh, D. (1994). The 1993 Walter Hubert Lecture: The role of the p53 tumor-suppressor gene in tumorigenesis. Br. J. Cancer 69: 409-416.
Livingstone, L.R., White, A., Sprouse, J., Livanos, E., Jacks, T. and Tisty, T.D. (1992). Altered cell cycle arrest and gene amplification potential accompany loss of wild-type p53. Cell 70: 932-935.
Meling, G.I., Lothe, R.A., Borresen, A.-L., Graue, C., Hauge, S., Clausen, O.P.F. and Rognum, T.O. (1993). The TP53 tumor suppressor gene in colorectal carcinomas. II. Relation to DNA ploidy pattern and clinicopathological variables. Br. J. Cancer 67: 93-98.
Murakami, Y., Hayashi, K. and Seikiya, T. (1991). Detection of aberrations of the p53 alleles and the gene transcript in human tumor cell lines by Single-Strand Conformation Polymorphism analysis. Cancer Res. 52: 3356-3361.
Nagai, M.A., Pacheco, M.M., Brentani, M.M., Marques, L.A., Brentani, R.R., Ponder, B.A.J. and Mulligan, L.M. (1994). Allelic loss on distal chromosome 17p is associated with poor prognosis in a group of Brazilian breast cancer patients. Br. J. Cancer 69: 754-758.
Nakamura, Y., Ballard, L., Leppert, M., O'Connell, P., Lathrop, G.M., Lalouel, J.M. and White, R. (1988). Isolation and mapping of a polymorphic DNA sequence pYNZ22 on chromosome 17p (D17S30). Nucleic Acids Res. 16: 5707.
Remvikos, Y., Tominaga, O., Hammel, P., Laurent-Puig, P., Salmon, R.J., Dutrillax, B. and Thomas, G. (1992). Increased p53 protein content of colorectal tumours correlates with poor survival. Br. J. Cancer 66: 758-764.
Rodrigues, N.R., Rowan, A., Smith, M.E.F., Kerr, I.B., Bodmer, W.F., Gannon, J.V. and Lane, D.P. (1990). p53 mutations in colorectal cancer. Proc. Natl. Acad. Sci. USA 87: 7555-7559.
Scott, N., Sagar, P., Stewart, J., Blair, G.E., Dixon, M.F. and Quirke, P. (1991). p53 in colorectal cancer: clinico-pathological correlation and prognostic significance. Br. J. Cancer 63: 317-319.
Soussi, T., Legros, Y., Lubin, R., Ory, K. and Schlichtholz, B. (1994). Multifactorial analysis of p53 alteration in human cancer: A review. Int. J. Cancer 57: 1-9.
Tornaletti, S. and Pfeifer, G.P. (1995). Complete and tissue-independent methylation of CpG sites in the p53 gene: implications for mutations in human cancers. Oncogene 10: 1493-1499.
Wang, L., Patel, U., Ghosh, L., Chen, H.C. and Banerjee, S. (1993). Mutation in the nm23 gene is associated with metastasis in colorectal cancer. Cancer Res. 53: 717-720.
Weber, J.L., Kwitek, A.E., May, P.E., Wallace, M.R., Collins, F.S. and Ledberter, H.D. (1990). Dinucleotide repeat polymorphisms at the D17S250 and D17S261 loci. Nucleic Acids Res 18: 4640.
Weissenbach, J., Gyapay, G., Dib, C., Vignal, A., Morissette, J., Millasseau, P., Vaysseix, G.A. and Lathrop, M. (1992). A second generation linkage map of the human genome. Nature 359: 794-801.
Yin, Y., Tainski, M.A., Bischoff, F.Z., Strong, L.C. and Wahl, G.M. (1992). Wild-type p53 restores cell cycle control and inhibits gene amplification in cells with mutant p53 alleles. Cell 70: 937-948.

Publication Dates

Publication in this collection
26 Oct 2006
Date of issue
1996

This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.

[1] Baker, S.J., Preisinger, A.C., Jessup, J.M., Paraskeva, C., Markowitz, S., Wilson, J.K.V., Hamilton, S. and Vogelstein, B. (1990). p53 gene mutations occur in combination with 17p allelic deletions as late events in colorectal tumorigenesis. Cancer Res. 50: 7717-7722.

[2] Bartek, J., Bartkova, J., Vojtesek, B., Stakova, Z., Lukas, J., Rejthar, A., Kovarik, J., Midgley, C.A., Gannon, J.V. and Lane, D.P. (1991). Aberrant expression of the p53 oncoprotein is a common feature of a wide spectrum of human malignancies. Oncogene 6: 1699-1703.

[3] Bedi, A., Pasricha, P.J., Akhtar, A.J., Barber, J.P., Bedi, G.C., Giardiello, F.M., Zehnbauer, S.R. and Jones, R.J. (1995). Inhibition of apoptosis during development of colorectal cancer. Cancer Res. 55: 1811-1816.

[4] Bosari, S., Viale, G., Bossi, P., Maggioni, M., Coggi, G., Murray, J.J. and Lee, A.K.C. (1994). Cytoplasmic accumulation of p53 protein: an independent prognostic indicator in colorectal adenomas. J. Natl. Cancer Inst. 86: 681-687.

[5] Cho, Y., Gorina, S., Jeffrey, P.D. and Pavletich, N.P. (1994). Crystal structure of a p53 tumor suppressor-DNA complex: Understanding tumorigenic mutations. Science 265: 346-355.

[6] Cohn, K.H., Wang, F., DeSoto-Lapaix, F., Solomon, W.B., Patterson, L.G., Arnold, M.R., Wemar, J., Feldman, J.G., Levy, A.T., Leone, A. and Steeg, P.S. (1991). Association of nm23-H1 allelic deletions with distant metastases in colorectal carcinoma. Lancet 338: 722-724.

[7] Coulondre, C., Miller, J.H., Farabaugh, P.J. and Gilbert, W. (1978). Molecular basis of base substitution hotspots in Escherichia coli. Nature 274: 775-780.

[8] Cunningham, J., Lust, J.A., Schaid, D.J., Bren, G.D., Carpenter, H.A., Rizza, E., Kovach, J.S. and Thibodeau, S.N. (1992). Expression of p53 and 17p allelic loss in colorectal carcinoma. Cancer Res. 52: 1974-1980.

[9] Eyfjord, J.E., Thorlacius, S., Steinarsdottir, M., Valgardsdottir, R., Ogmundsdottir, H.M. and Anamthawat-Jonsson, K. (1995). p53 abnormalities and genomic instability in primary human breast carcinomas. Cancer Res. 55: 646-651.

[10] Fearon, E.R. and Vogelstein, B. (1990). A genetic model for colorectal tumorigenesis. Cell 61: 759-767.

[11] Finlay, C.A., Hinds, P.W., Tan, T.-H., Eliyahu, D., Oren, D. and Levine, A.J. (1988). Activating mutations for transformation by p53 produce a gene product that forms an hsc70-p53 complex with an altered half-life. Mol. Cell. Biol. 8: 531-539.

[12] Futreal, P.A., Barret, J.C. and Weiseman, R.W. (1991). An Alu polymorphism intragenic to the TP53 gene. Nucleic Acids Res. 19: 6977.

[13] Gannon, J.V., Greaves, R., Iggo, R. and Lane, D.P. (1990). Activating mutations in p53 produce a common conformational effect. A monoclonal antibody specific for the mutant form. EMBO J. 9: 1595-1602.

[14] Goh, H.-S., Chan, C.-S., Khine, K. and Smith, D.R. (1994). p53 and behaviour of colorectal cancer. Lancet 344: 233-234.

[15] Hall, J.M., Friedman, L., Guenther, C., Lee, M.K., Weber, J.L., Black, D.M. and King, M.-C. (1992). Closing in on a breast cancer gene on chromosome 17q. Am. J. Hum. Genet. 50: 1235-1242.

[16] Harris, C.C. and Hollstein, M. (1993). Clinical implications of the p53 tumor suppressor gene. N. Engl. J. Med. 329: 1318-1327.

[17] Hasegawa, H., Ueda, M., Furukawa, K., Watanabe, M., Teramoto, T., Mukai, M. and Kitajima, M. (1995). p53 gene mutations in early colorectal carcinoma. De novo vs. adenoma-carcinoma sequence. Int. J. Cancer 64: 47-51.

[18] Hoffman, B. and Liebermann, D.A. (1994). Molecular controls of apoptosis: differentiation/growth arrest primary response genes, proto-oncogenes, and tumor suppressor genes as positive and negative modulators. Oncogene 9: 1807-1812.

[19] Hollstein, M., Sidransky, D., Vogelstein, B. and Harris, C.C. (1991). p53 in human cancers. Science 253: 49-53.

[20] Jones, M.H. and Nakamura, Y. (1992). Detection of loss of heterozygosity at the human TP53 locus using a dinucleotide repeat. Genes Chromosom. Cancer 5: 89-90.

[21] Khine, K., Smith, D.R. and Goh, H.-S. (1993). High frequency of allelic deletion on chromosome 17p in advanced colorectal cancer. Cancer 73: 28-35.

[22] Kikuchi-Yanoshita, R., Konishi, M., Ito, S., Seki, M., Tanaka, K., Maeda, Y., Iino, H., Fukayama, M., Koike, M., Mori, T., Sakamura, H., Fukunari, H., Iwama, T. and Miyaki, M. (1992). Genetic changes of both p53 alleles associated with the conversion from colorectal adenoma to early carcinoma in familial adenomatous polyposis and non-familial adenomatous polyposis patients. Cancer Res. 52: 3965-3967.

[23] Knudson, A.G. (1971). Mutation and cancer: Statistical study of retinoblastoma. Proc. Natl. Acad. Sci. USA 68: 820-823.

[24] Lane, D.P. (1994). The regulation of p53 function: Steiner award lecture. Int. J. Cancer 57: 623-627.

[25] Levine, A.J., Perry, M.P., Chang, A., Silver, A., Dittmer, D., Wu, M. and Welsh, D. (1994). The 1993 Walter Hubert Lecture: The role of the p53 tumor-suppressor gene in tumorigenesis. Br. J. Cancer 69: 409-416.

[26] Livingstone, L.R., White, A., Sprouse, J., Livanos, E., Jacks, T. and Tisty, T.D. (1992). Altered cell cycle arrest and gene amplification potential accompany loss of wild-type p53. Cell 70: 932-935.

[27] Meling, G.I., Lothe, R.A., Borresen, A.-L., Graue, C., Hauge, S., Clausen, O.P.F. and Rognum, T.O. (1993). The TP53 tumor suppressor gene in colorectal carcinomas. II. Relation to DNA ploidy pattern and clinicopathological variables. Br. J. Cancer 67: 93-98.

[28] Murakami, Y., Hayashi, K. and Seikiya, T. (1991). Detection of aberrations of the p53 alleles and the gene transcript in human tumor cell lines by Single-Strand Conformation Polymorphism analysis. Cancer Res. 52: 3356-3361.

[29] Nagai, M.A., Pacheco, M.M., Brentani, M.M., Marques, L.A., Brentani, R.R., Ponder, B.A.J. and Mulligan, L.M. (1994). Allelic loss on distal chromosome 17p is associated with poor prognosis in a group of Brazilian breast cancer patients. Br. J. Cancer 69: 754-758.

[30] Nakamura, Y., Ballard, L., Leppert, M., O'Connell, P., Lathrop, G.M., Lalouel, J.M. and White, R. (1988). Isolation and mapping of a polymorphic DNA sequence pYNZ22 on chromosome 17p (D17S30). Nucleic Acids Res. 16: 5707.

[31] Remvikos, Y., Tominaga, O., Hammel, P., Laurent-Puig, P., Salmon, R.J., Dutrillax, B. and Thomas, G. (1992). Increased p53 protein content of colorectal tumours correlates with poor survival. Br. J. Cancer 66: 758-764.

[32] Rodrigues, N.R., Rowan, A., Smith, M.E.F., Kerr, I.B., Bodmer, W.F., Gannon, J.V. and Lane, D.P. (1990). p53 mutations in colorectal cancer. Proc. Natl. Acad. Sci. USA 87: 7555-7559.

[33] Scott, N., Sagar, P., Stewart, J., Blair, G.E., Dixon, M.F. and Quirke, P. (1991). p53 in colorectal cancer: clinico-pathological correlation and prognostic significance. Br. J. Cancer 63: 317-319.

[34] Soussi, T., Legros, Y., Lubin, R., Ory, K. and Schlichtholz, B. (1994). Multifactorial analysis of p53 alteration in human cancer: A review. Int. J. Cancer 57: 1-9.

[35] Tornaletti, S. and Pfeifer, G.P. (1995). Complete and tissue-independent methylation of CpG sites in the p53 gene: implications for mutations in human cancers. Oncogene 10: 1493-1499.

[36] Wang, L., Patel, U., Ghosh, L., Chen, H.C. and Banerjee, S. (1993). Mutation in the nm23 gene is associated with metastasis in colorectal cancer. Cancer Res. 53: 717-720.

[37] Weber, J.L., Kwitek, A.E., May, P.E., Wallace, M.R., Collins, F.S. and Ledberter, H.D. (1990). Dinucleotide repeat polymorphisms at the D17S250 and D17S261 loci. Nucleic Acids Res 18: 4640.

[38] Weissenbach, J., Gyapay, G., Dib, C., Vignal, A., Morissette, J., Millasseau, P., Vaysseix, G.A. and Lathrop, M. (1992). A second generation linkage map of the human genome. Nature 359: 794-801.

[39] Yin, Y., Tainski, M.A., Bischoff, F.Z., Strong, L.C. and Wahl, G.M. (1992). Wild-type p53 restores cell cycle control and inhibits gene amplification in cells with mutant p53 alleles. Cell 70: 937-948.