人胃癌组织WAF1基因的缺失与不表达

探讨ＷＡＦ１与胃癌的关系，为胃癌基因治疗提供有价值的依据。方法３２例胃癌和３０例正常胃粘膜标本提取细胞ＤＮＡ、总ＲＮＡ，ＭＴＳ１ｃＤＮＡ探针采用随机引物法进行３２Ｐ－ｄＡＴＰ掺入标记，ＷＡＦ１与所提ＤＮＡ、ＲＮＡ进行Ｓｏｕｔｈｅｒｎ和Ｎｏｒｔｈｅｒｎ杂交分析，检测标本中有无ＷＡＦ１的缺失和ｍＲＮＡ的不表达。ｐ５３抗体与提取的蛋白质进行Ｗｅｓｔｅｒｎ印迹分析，检测标本中有无ｐ５３的表达。结果ＷＡＦ１Ｓｏｕｔｈｅｒｎ和Ｎｏｒｔｈｅｒｎ杂交分析结果显示，３２例胃癌标本ＷＡＦ１基因缺失率及ｍＲＮＡ不表达率分别为３４．４％和４０．６％，明显高于正常胃粘膜组。ＷＡＦ１基因缺失率及ｍＲＮＡ不表达与胃癌浸润的深度及分化程度有关，胃癌侵及肌层组ＷＡＦ１基因缺失及ｍＲＮＡ不表达率分别为５２．６％和５７．９％，胃癌未侵及肌层组分别为７．７％和１５．４％，高、中分化胃癌组ＷＡＦ１基因缺失及ｍＲＮＡ不表达率分别为７．１％和１４．３％，而低分化及印戒细胞胃癌组分别为５５．８％和６１．１％。结论胃癌的发生与ｐ５３失活及ＷＡＦ１ｍＲＮＡ不表达有关，胃癌组织中ｐ５３失活与ＷＡＦ１ｍＲＮＡ不表达关系密切。

RELATIONSHIP OF WAF1 AND p53 IN HUMAN GASTRIC CARCINOMAS

PURPOSE To observe expression of WAF1 and p53 gene in human gastric carcinomas. METHODS The totals RNA and proteins were isolated from samples of 32 gastric cancers. WAF1 cDNA probes were labeled by random priming with 32 P-dATP. Northern blot was used to detect whether WAF1 mRNA unexpressed in the samples. The sample protein was analyzed by Western bolt with p53 antibody in order to whether p53 expressed. RESULTS WAF1 Northern blot displayed that 13 patients were negative in 32 gastric cancers. p53 of 15 cases in 32 gastric cancers were positive by Western bolt with p53 antibody. Among 15 patients with gastric cancer who were positive reaction of Western bolt with mutant p53 antibody, 13 patients had unexpression of WAF1 mRNA. Nobady had expression of WAF1 mRNA. Nobady had expression of WAF1 mRNA among 17 patients who were negative reaction of Western bolt with mutant p53 antibody. CONCLUSIONS The occurrence of gastric cancer was related to unexpression of WAF1 mRNA and inactive of p53. Inactivation of p53 was closely related to unexpression of WAF1 mRNA.