p57～（kip2）在人体肝癌中杂合性缺失及mRNA和蛋白表达关系研究
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摘要
目的：研究P57~(kip2)基因在人体肝细胞癌(Hepatocellular carcinoma，HCC)不同阶段的杂合性缺失(Loss of heterozygosity，LOH)及其mRNA和蛋白表达情况，以及三者之间的关系，以探讨HCC发生的分子生物学机制。
方法：运用PCR—聚丙烯酰胺凝胶电泳—银染法对30例肝癌组织与P57~(kip2)基因同一区域的3个微卫星位点(D11S1397，D11S1318和D11S4046)进行LOH的检测，同时运用原位分子杂交和免疫组化技术分别检测30例正常肝组织、30例癌周肝硬化和30例肝癌组织中P57~(kip2)mRNA及P57~(kip2)蛋白的表达情况。
结果：1．LOH检测：30例肝癌组织中在3个微卫星位点均未出现LOH。2．P57~(kip2)mRNA表达：正常肝组织未见表达，癌周肝硬化组与肝癌组阳性表达率为26.7％(8／30)。肝癌组分化好和分化差阳性表达率之间差异无统计学意义。3．P57~(kip2)蛋白表达：正常肝组织未见表达，癌周肝硬化组与肝癌组阳性表达率分别为56.67％(17／30)和63.33％(19／30)。肝癌组分化好者与分化差者阳性表达率之间差异无统计学意义。4．肝癌3个微卫星位点LOH与P57~(kip2)mRNA和蛋白表达均无关联性。5．肝癌P57~(kip2)mRNA与蛋白表达之间有关联性。
结论：1．P57~(kip2)mRNA和蛋白表达异常提示其可能在原发性HCC的发生中起重要作用。2．与P57~(kip2)基因同一区域的3个微卫星位点未出现LOH，提示P57~(kip2)表达异常与该3个微卫星位点无关，可能存在其它机制。
Objective: To study the loss of hererozygosity(LOH) of P57~(kip2) and expressions of P57~(kip2) mRNA & protein in human hepatocellular carcinoma (HCC) in order to explore the molecular mechanism of hepatocarcinogenesis.
Methods: LOH of three microsatellite loci(D11S1397 D11S1318 and D11S4046) were detected where the P57~(kip2) gene is in by using PCR-Polyacrylamide gel electrophoresis - silver staining method in 30 cases of HCC. Immunohistochemistry staining and in situ hybridization (ISH) were also used to show the expression of P57~(kip2) protein and mRNA in pericancerous cirrhosis and HCC.
Results: 1. LOH detected: LOH was not identified in 30 cases on three microsatellite locies. 2. Expression of P57~(kip2) mRNA: There were no expressions in normal liver tissue. Expressions both in pericancerous cirrhosis and hepatocellular carcinoma were 26. 67%(8/30). There was no significant difference between the expressions in high differentiated HCC tissues and low differentiated HCC tissues. 3. Expression of P57~(kip2) protein: There were no expressions in normal liver tissue. Expressions in precancerous cirrhosis and hepatocellular carcinoma were 56. 67% (17/30) and 63.33 %( 19/30) respectively. There was no significant difference between the expressions in high differentiated HCC tissues and low differentiated HCC tissues. 4. There was no correlation between the LOH of three locies and the expression of P57~(kip2)mRNA; there was no correlation between the LOH of three locies and the expression of P57~(kip2) protein. 5. There was positive correlation between expression of P57~(kip2) mRNA and expression of P57~(kip2) protein.
Conclusion: 1. The disorder expressions of P57~(kip2) mRNA and protein indicated that P57~(kip2) may be involved in hepatocarcinogenesis and prognosis. 2. LOH was not identified on three microsatellite locies , which indicated that there may be another reason for the disorder expression of P57~(kip2).
方法：运用PCR—聚丙烯酰胺凝胶电泳—银染法对30例肝癌组织与P57~(kip2)基因同一区域的3个微卫星位点(D11S1397，D11S1318和D11S4046)进行LOH的检测，同时运用原位分子杂交和免疫组化技术分别检测30例正常肝组织、30例癌周肝硬化和30例肝癌组织中P57~(kip2)mRNA及P57~(kip2)蛋白的表达情况。
结果：1．LOH检测：30例肝癌组织中在3个微卫星位点均未出现LOH。2．P57~(kip2)mRNA表达：正常肝组织未见表达，癌周肝硬化组与肝癌组阳性表达率为26.7％(8／30)。肝癌组分化好和分化差阳性表达率之间差异无统计学意义。3．P57~(kip2)蛋白表达：正常肝组织未见表达，癌周肝硬化组与肝癌组阳性表达率分别为56.67％(17／30)和63.33％(19／30)。肝癌组分化好者与分化差者阳性表达率之间差异无统计学意义。4．肝癌3个微卫星位点LOH与P57~(kip2)mRNA和蛋白表达均无关联性。5．肝癌P57~(kip2)mRNA与蛋白表达之间有关联性。
结论：1．P57~(kip2)mRNA和蛋白表达异常提示其可能在原发性HCC的发生中起重要作用。2．与P57~(kip2)基因同一区域的3个微卫星位点未出现LOH，提示P57~(kip2)表达异常与该3个微卫星位点无关，可能存在其它机制。
Objective: To study the loss of hererozygosity(LOH) of P57~(kip2) and expressions of P57~(kip2) mRNA & protein in human hepatocellular carcinoma (HCC) in order to explore the molecular mechanism of hepatocarcinogenesis.
Methods: LOH of three microsatellite loci(D11S1397 D11S1318 and D11S4046) were detected where the P57~(kip2) gene is in by using PCR-Polyacrylamide gel electrophoresis - silver staining method in 30 cases of HCC. Immunohistochemistry staining and in situ hybridization (ISH) were also used to show the expression of P57~(kip2) protein and mRNA in pericancerous cirrhosis and HCC.
Results: 1. LOH detected: LOH was not identified in 30 cases on three microsatellite locies. 2. Expression of P57~(kip2) mRNA: There were no expressions in normal liver tissue. Expressions both in pericancerous cirrhosis and hepatocellular carcinoma were 26. 67%(8/30). There was no significant difference between the expressions in high differentiated HCC tissues and low differentiated HCC tissues. 3. Expression of P57~(kip2) protein: There were no expressions in normal liver tissue. Expressions in precancerous cirrhosis and hepatocellular carcinoma were 56. 67% (17/30) and 63.33 %( 19/30) respectively. There was no significant difference between the expressions in high differentiated HCC tissues and low differentiated HCC tissues. 4. There was no correlation between the LOH of three locies and the expression of P57~(kip2)mRNA; there was no correlation between the LOH of three locies and the expression of P57~(kip2) protein. 5. There was positive correlation between expression of P57~(kip2) mRNA and expression of P57~(kip2) protein.
Conclusion: 1. The disorder expressions of P57~(kip2) mRNA and protein indicated that P57~(kip2) may be involved in hepatocarcinogenesis and prognosis. 2. LOH was not identified on three microsatellite locies , which indicated that there may be another reason for the disorder expression of P57~(kip2).
引文
1. Simonetti RG, Camma C, Fiorello F, et al. Hepatocellular carcinoma. A worldwide problem and the major risk factors. Dig Dis Sci, 1991, 36:962-972.
2. Barbacid T, Sanuits W, Civin C, et al. P53 gene mRNA expression and chromosome 17p allele loss in hepatocellular carcinomas. J Chin Oncol, 1996,14(8):2224-2231
3. Reid LH, Crider Miller SJ, Ande West, et al. Genomic organization of the human p57~(kip2) gene and its analysis in the G401 Wilms' Tumor assay. Cancer Res, 1996, 56(6): 1214
4. Matsuka S, Thompson J, Edwards M, et al. Imprinting of the gene encoding a human Cyclin-dependent kinase inhibitor p57~(Kip2)on chromosome 11p15.5. Proc Natl Acad Sci USA, 1996, 93(8): 3026-3030.
5. Kawamata N, Morosetti R, Miller CW, et al. Molecular analysis of the cyclin-dependent kinase inhibitor gene p27/Kipl in human malignancies. Cancer Res, 1995, 55(11): 2266-2269.
6. Russo AA, Jeffrey PD, Patten AK, et al. Crystal structure of the p27Kipl cyclin-dependent-kinase inhibitor bound to the cyclin A-Cdk2 complex. Nature, 1996, 382(6589): 325-331.
7. Luo Y, Hurwitz J, Massagen J, et al. Cell-cyclin inhibition by independent CDK and PCNA binding domains in p21Cipl. Nature, 1995, 375(6527): 159-161.
8. Reid LH, Crider Miller SJ, hnde West, et al. Genomic organization of the human p57kip2 gene and its analysis in the G401Wilms' Tumor assay. Cancer Res, 1996, 56(6): 1214-1218
9. Rosenberg E, Demopoulos RI, Jacquotte AZ, et al. Expression of cell cycle regulation regulators p57~(kip2), CyclinD_1 and CyclinE in epithelial ovarian tumors and survival. Hum Pathol 2001; 32:808-813.
10. Zhang PM, Liegeosis NJ, Wong C, et al. Altered cell differentiation and proliferation in mice lacking p57~(kip2) indicates a role in Beckwith-Wiedemann syndrome. Nature, 1997,387:151
11．曹守强等。p57~(kip2)蛋白在进展期胃癌中的表达及临床意义。哈尔滨医科大学学报2004．38(6)：561-563
12. Ito Y, takeda T, Wakasa k, et al. Expression of p57~(kip2) protein in pancreatic adenocarcinoma. Pancreas. 2001,23(3):246-256
13. Noura S, Yamamoto H, Sekimoto M, et al. Expression of second class of KIP protein p57~(kip2) in human colorectal carcinoma. Int J Oncol, 2001, 1991(1):39-47
14．p57~(kip2)和cyclinE在人宫颈癌中的表达及其与临床病理及预后的关系。第四军医大学学报，2006，27(15)：1434-1437
15．邹琼等。人类肝癌发生过程中CyclinD_1和CDK_4及P16蛋白的表达。肿瘤防治杂志，2005，12(15)：1127-113l
16．孔萌萌。Rb与p57~(kip2)在肝细胞癌发生发展过程中的表达研究：[硕士学位论文]。昆明医学院：基础医学院，2004。
17．周峥珍。DEN诱发大鼠肝癌的病理学及细胞周期调控因子表达的研究：[硕士学位论文]。昆明医学院：基础医学院，2005。
18．郭卉等。p57~(kip2)在肝癌组织中的表达及其于临床病理因素、PCNA和P53的关系。第四军医大学学报，2004，25(17)：1562-1565
19. Nakai S, Maszki T, Shiratori Y, et al. Expression of p57kip2 in hepatocellular carcinoma: relationship between tumor differentiation and patient survival. Int Oncol, 2002, 20(4): 769-775
20. Stewart MC, Kadlcek RM, Robbins PD, et al. Expression and activity of the CDK inhibitor p57Kip2 in chondrocytes undergoing hypertrophic differentiation. Bone Miner Res. 2004 Jan; 19(1):123-32.
21. Wang S, Kim JH, Moon KC, et al. Cell-cycle mechanisms involved in podocyte proliferation in cellular lesion of focal segmental glomerulosclerosis. Am J Kidney Dis. 2004 Jan; 43(1): 19-27.
22. Hoffmann MJ, Florl AR, Seifert HH, et al. Multiple mechanisms downregulate CDKNIC in human bladder cancer. Int J Cancer, 2005 Apt 10; 114(3): 406-413.
23. Christine Sempoux, Yves Guiot, Karin Dahan, et al. The focal form of persistent hyperinsulinemic hypoglycemia of infancy. Diabetes, 52:784-794, 2003
24. Syeling Lai, Helmuth Goepfert, Ann M. Gillenwater, et al. Loss of Imprinting and Genetic Alterationsof the Cyclin-dependent Kinase Inhibitor p57KIP2 Gene in Head and Neck Squamous Cell Carcinoma. Clinical Cancer Research, 2000, 6(8), 3172-3176.
25. Kassem SA, Ariel I, Thornton PS ,etal. p57(KIP2) expression in normal islet cells and in hyperinsulinism of infancy. Diabetes, 2001,50: 2763-2769.
26. Bourcigaux N, Gaston V, Logie A etal. High expression of cyclin E and G1 CDK and loss of function of p57KIP2 are involved in proliferation of malignant sporadic adrenocortical tumors. Clin Endocrinol Metab, 2000,85(1):322-327.
27. Esteller M, Levine R, Baylin SB, et aI. MLHI promoter hypermethylation is associ-ated with the microsatellite instability phenotype in sporadic endometrial carcinomas PMID,1998, IT:2413-2417
28. Macdonald GA, Green son TK, Ssitok, et al. Microsatellite instability ad loss of hetero zygosity at DNA mismatch repair gene loci occurs during hepatic carcinogenesis. Hepatology, 199828(1):90-97
1. Loeb LA. A mutator phenotype in cancer. Cancer Res, 2001,61(8):3230-3239
2．丛文铭。肿瘤抑制基因变异的分子，病理学研究进展中华病理学杂志，2001，30(1)：58-60
3. Bocker T, Diermann J, Friedl W, et al. Microsatellite instability analysis: a multicenter study for reliability and quality control. Cancer Res, 1997, 57:4739-4743
4. Fishel, LescoeMK, Rao MRS, et al .The human mutation gene homology MSHZ and its association witb hereditary monpolyposis colon cancer. Cell, 1993, 75(5): 1027
5. Esteller M, Levine R, Baylin SB, et al. MLHlpromoter hypermethylation is associated with the microsatellite instability phenotype in sporadic endometrial carcinomas .PMID, 1998,17:2413-2417
6．邓锡云、雷亚，基因组不稳定性与肿瘤，国外遗传学分册，1997，20：1621
7. Muta H, Moguchi M, perucho M, et al. Clinical implication of microsatellite instability in cdorectal cancers .cancer ,1996, T:265-270
8. Take mura S, Yashiro M, sunami T, et al. Novel models for human scirrhous gastric carcinoma in vivo. Cancer sci, 2004, 95:893-900
9．王永信，柯杨，宁涛等。中国人胃癌微卫星DNA的不稳定性研究。中华医学遗传学杂志，1998，15：155-157
10. kazachkov Y, Yoffe B, khaoustov VI et al .Liver ,1998;18(3):156-161
11. Macdonald GA, Green son JK, et al. Microsatellite instability ad loss of heterozygosity at DNA mismatch repair gene loci occurs during hepatic carcinogenesis. Hepatology, 1998,28(1): 90-97
12. CODEGONI A M, BERTONI F, COLELLA G, et al. Microsatellite instability and frameshift mutations in genes involved in cell cycle progression or apoptosis in ovarian cancer. Oncol Res, 1999, 11(7): 297
13. KOBAYASHI K, SAGAE S, KUDOR, et al. Microsatellite instability in endometrial carcinomas:frequent replication errors in tumors of early onset and/or of poorly differentiated type. Genes chromosomes cancer, 1995, 14(2):128
14. Fujita M, Enomoto T, Yoshinok ,et al.Microsatellite instability and alterations in the hMSH2 gene in human ovarian cancer . Int J Cancer, 1995;64(6):361-366
15. Mao L, Schoenberg MP, scicchitano M, et al ,science ,1996,271:659-662
16. Ramon Parsons, Guo min Li ,Matthew J, et al. Cell ,1993;75:1227-1236
17. Reitmair AH, schmits R, Ewel A ,et al. Nat Genet, 1995;11:64-70
18. Peltomaki P, Aaltonen LA, Sistonen P, et al.Gentic mapping of a locus predisposing to human colorectal cancer. Science, 1993,260:810-812
19. Rushoff T, Bocker J, schlegel T, et al. Microsatellite instability:new aspect in the carcinogenesis of colorectal carcinoma .Virchow Arch, 1995,426:215-222
20. Wooster R, Bignell G, Lancaster JA, et al ,Identification of the breast cancer susceptibilisy gene BRCA2. Nature ,1995,378:789-792
21. Hahn SA, schutte M, hogue AT, et al. DPC4 a candidate tumor suppressor gene at human chromosome 18q21.1 .Science ,1996,271:350-353
22. Li YL, Tian Z, Wu DY, et al. Loss of heterozygosity on 10q23.3 and mutation of tumor,suppressor gene PTEN in gastric cancer and precancerous lesions.World J Gastroenterol, 2005 11(2):285-288
23．于观贞等，P53和nm23杂合性缺失及表达异常在晚期胃肠癌转移中的作用。肿瘤，2006，26(8)：717-720
24．于颖彦等。胃癌5号染色体短臂杂合性缺失的精细作图及其相关基因探讨。中华肿瘤杂志，2006，28(2)：84-87
25．丛文铭，吴孟超，陈汉。肝内胆管癌多基因变异表型分析。中华医学杂志，2001，81(5)：271-273
26．丛文铭，Finkelstein SD，吴孟超。胆管癌基因变异与临床病理学特征的关系。中华病理学杂志，2001，30(3)：183-187
27. Cong WM, Bakker A, Swalsky PA, et al .Multiple genetic alterations involved in the tum origenesis of human cholangiocarcinoma:a molecular genetic and clinicopathological study .J Cancer Res Clin Oncol ,2001,127(3):187-192
28. Piao Z, kim H, Jeon BK ,et al. Pelationship between loss of heterozygosity of tumor suppressor genes and histologic differentiation in hepatocellular carcinoma. Cancer 1997,80:865-872
29．丛文铭等。肝细胞癌肿瘤抑制基因的杂合性缺失和微卫星不稳定性研究。中华病理学杂志，2005，(34)2：71-74
30. Elamidi A, Hamoudi RA, Kocjan G; et al.Cerrical intraepithelial neoplasia:prognosis by combined LOH analysis of multiple loci.Gynecol Oncol,2004,94(3):671-679
2. Barbacid T, Sanuits W, Civin C, et al. P53 gene mRNA expression and chromosome 17p allele loss in hepatocellular carcinomas. J Chin Oncol, 1996,14(8):2224-2231
3. Reid LH, Crider Miller SJ, Ande West, et al. Genomic organization of the human p57~(kip2) gene and its analysis in the G401 Wilms' Tumor assay. Cancer Res, 1996, 56(6): 1214
4. Matsuka S, Thompson J, Edwards M, et al. Imprinting of the gene encoding a human Cyclin-dependent kinase inhibitor p57~(Kip2)on chromosome 11p15.5. Proc Natl Acad Sci USA, 1996, 93(8): 3026-3030.
5. Kawamata N, Morosetti R, Miller CW, et al. Molecular analysis of the cyclin-dependent kinase inhibitor gene p27/Kipl in human malignancies. Cancer Res, 1995, 55(11): 2266-2269.
6. Russo AA, Jeffrey PD, Patten AK, et al. Crystal structure of the p27Kipl cyclin-dependent-kinase inhibitor bound to the cyclin A-Cdk2 complex. Nature, 1996, 382(6589): 325-331.
7. Luo Y, Hurwitz J, Massagen J, et al. Cell-cyclin inhibition by independent CDK and PCNA binding domains in p21Cipl. Nature, 1995, 375(6527): 159-161.
8. Reid LH, Crider Miller SJ, hnde West, et al. Genomic organization of the human p57kip2 gene and its analysis in the G401Wilms' Tumor assay. Cancer Res, 1996, 56(6): 1214-1218
9. Rosenberg E, Demopoulos RI, Jacquotte AZ, et al. Expression of cell cycle regulation regulators p57~(kip2), CyclinD_1 and CyclinE in epithelial ovarian tumors and survival. Hum Pathol 2001; 32:808-813.
10. Zhang PM, Liegeosis NJ, Wong C, et al. Altered cell differentiation and proliferation in mice lacking p57~(kip2) indicates a role in Beckwith-Wiedemann syndrome. Nature, 1997,387:151
11．曹守强等。p57~(kip2)蛋白在进展期胃癌中的表达及临床意义。哈尔滨医科大学学报2004．38(6)：561-563
12. Ito Y, takeda T, Wakasa k, et al. Expression of p57~(kip2) protein in pancreatic adenocarcinoma. Pancreas. 2001,23(3):246-256
13. Noura S, Yamamoto H, Sekimoto M, et al. Expression of second class of KIP protein p57~(kip2) in human colorectal carcinoma. Int J Oncol, 2001, 1991(1):39-47
14．p57~(kip2)和cyclinE在人宫颈癌中的表达及其与临床病理及预后的关系。第四军医大学学报，2006，27(15)：1434-1437
15．邹琼等。人类肝癌发生过程中CyclinD_1和CDK_4及P16蛋白的表达。肿瘤防治杂志，2005，12(15)：1127-113l
16．孔萌萌。Rb与p57~(kip2)在肝细胞癌发生发展过程中的表达研究：[硕士学位论文]。昆明医学院：基础医学院，2004。
17．周峥珍。DEN诱发大鼠肝癌的病理学及细胞周期调控因子表达的研究：[硕士学位论文]。昆明医学院：基础医学院，2005。
18．郭卉等。p57~(kip2)在肝癌组织中的表达及其于临床病理因素、PCNA和P53的关系。第四军医大学学报，2004，25(17)：1562-1565
19. Nakai S, Maszki T, Shiratori Y, et al. Expression of p57kip2 in hepatocellular carcinoma: relationship between tumor differentiation and patient survival. Int Oncol, 2002, 20(4): 769-775
20. Stewart MC, Kadlcek RM, Robbins PD, et al. Expression and activity of the CDK inhibitor p57Kip2 in chondrocytes undergoing hypertrophic differentiation. Bone Miner Res. 2004 Jan; 19(1):123-32.
21. Wang S, Kim JH, Moon KC, et al. Cell-cycle mechanisms involved in podocyte proliferation in cellular lesion of focal segmental glomerulosclerosis. Am J Kidney Dis. 2004 Jan; 43(1): 19-27.
22. Hoffmann MJ, Florl AR, Seifert HH, et al. Multiple mechanisms downregulate CDKNIC in human bladder cancer. Int J Cancer, 2005 Apt 10; 114(3): 406-413.
23. Christine Sempoux, Yves Guiot, Karin Dahan, et al. The focal form of persistent hyperinsulinemic hypoglycemia of infancy. Diabetes, 52:784-794, 2003
24. Syeling Lai, Helmuth Goepfert, Ann M. Gillenwater, et al. Loss of Imprinting and Genetic Alterationsof the Cyclin-dependent Kinase Inhibitor p57KIP2 Gene in Head and Neck Squamous Cell Carcinoma. Clinical Cancer Research, 2000, 6(8), 3172-3176.
25. Kassem SA, Ariel I, Thornton PS ,etal. p57(KIP2) expression in normal islet cells and in hyperinsulinism of infancy. Diabetes, 2001,50: 2763-2769.
26. Bourcigaux N, Gaston V, Logie A etal. High expression of cyclin E and G1 CDK and loss of function of p57KIP2 are involved in proliferation of malignant sporadic adrenocortical tumors. Clin Endocrinol Metab, 2000,85(1):322-327.
27. Esteller M, Levine R, Baylin SB, et aI. MLHI promoter hypermethylation is associ-ated with the microsatellite instability phenotype in sporadic endometrial carcinomas PMID,1998, IT:2413-2417
28. Macdonald GA, Green son TK, Ssitok, et al. Microsatellite instability ad loss of hetero zygosity at DNA mismatch repair gene loci occurs during hepatic carcinogenesis. Hepatology, 199828(1):90-97
1. Loeb LA. A mutator phenotype in cancer. Cancer Res, 2001,61(8):3230-3239
2．丛文铭。肿瘤抑制基因变异的分子，病理学研究进展中华病理学杂志，2001，30(1)：58-60
3. Bocker T, Diermann J, Friedl W, et al. Microsatellite instability analysis: a multicenter study for reliability and quality control. Cancer Res, 1997, 57:4739-4743
4. Fishel, LescoeMK, Rao MRS, et al .The human mutation gene homology MSHZ and its association witb hereditary monpolyposis colon cancer. Cell, 1993, 75(5): 1027
5. Esteller M, Levine R, Baylin SB, et al. MLHlpromoter hypermethylation is associated with the microsatellite instability phenotype in sporadic endometrial carcinomas .PMID, 1998,17:2413-2417
6．邓锡云、雷亚，基因组不稳定性与肿瘤，国外遗传学分册，1997，20：1621
7. Muta H, Moguchi M, perucho M, et al. Clinical implication of microsatellite instability in cdorectal cancers .cancer ,1996, T:265-270
8. Take mura S, Yashiro M, sunami T, et al. Novel models for human scirrhous gastric carcinoma in vivo. Cancer sci, 2004, 95:893-900
9．王永信，柯杨，宁涛等。中国人胃癌微卫星DNA的不稳定性研究。中华医学遗传学杂志，1998，15：155-157
10. kazachkov Y, Yoffe B, khaoustov VI et al .Liver ,1998;18(3):156-161
11. Macdonald GA, Green son JK, et al. Microsatellite instability ad loss of heterozygosity at DNA mismatch repair gene loci occurs during hepatic carcinogenesis. Hepatology, 1998,28(1): 90-97
12. CODEGONI A M, BERTONI F, COLELLA G, et al. Microsatellite instability and frameshift mutations in genes involved in cell cycle progression or apoptosis in ovarian cancer. Oncol Res, 1999, 11(7): 297
13. KOBAYASHI K, SAGAE S, KUDOR, et al. Microsatellite instability in endometrial carcinomas:frequent replication errors in tumors of early onset and/or of poorly differentiated type. Genes chromosomes cancer, 1995, 14(2):128
14. Fujita M, Enomoto T, Yoshinok ,et al.Microsatellite instability and alterations in the hMSH2 gene in human ovarian cancer . Int J Cancer, 1995;64(6):361-366
15. Mao L, Schoenberg MP, scicchitano M, et al ,science ,1996,271:659-662
16. Ramon Parsons, Guo min Li ,Matthew J, et al. Cell ,1993;75:1227-1236
17. Reitmair AH, schmits R, Ewel A ,et al. Nat Genet, 1995;11:64-70
18. Peltomaki P, Aaltonen LA, Sistonen P, et al.Gentic mapping of a locus predisposing to human colorectal cancer. Science, 1993,260:810-812
19. Rushoff T, Bocker J, schlegel T, et al. Microsatellite instability:new aspect in the carcinogenesis of colorectal carcinoma .Virchow Arch, 1995,426:215-222
20. Wooster R, Bignell G, Lancaster JA, et al ,Identification of the breast cancer susceptibilisy gene BRCA2. Nature ,1995,378:789-792
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