摘要
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。
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.
出处
《上海医科大学学报》
CSCD
1998年第6期416-418,444,共4页
Journal of Fudan University(Medical Science)