【摘要】 目的 检测肿瘤抑制基因p16在胰腺腺癌细胞系中的基因改变和在mRNA以及蛋白质水平的表达。方法 使用PCR方法检测基因的缺失情况;使用单链构象多态性（SSCP）方法检测基因有无突变;使用甲基化特异性PCR（MSP）方法检测基因启动子区域的甲基化状态;使用RT-PCR方法检测基因在mRNA水平的表达;使用免疫细胞化学染色和Western blot方法检测基因在蛋白质水平的表达。结果 被检测的6个胰腺腺癌细胞系中,有3个细胞系存在p16基因的纯合性缺失,这3个细胞系在mRNA水平和蛋白质水平均无p16基因的表达。在全部6个细胞系中没有发现p16基因突变和启动子区域的过甲基化。结论 在被检测的6个胰腺腺癌细胞系中,纯合性缺失是造成p16基因失表达的原因。更多还原

【Abstract】 Objective To detect the gene alteration and expression in mRNA and protein levels of tumor suppressor gene p16 in pancreatic adenocarcinoma cell lines. Methods The deletion and mutation of the p16 gene were detected by PCR and SSCP; the methylation status of the promoter region of the p16 gene was detected by methylation-pecific PCR(MSP); the expression of p16 mRNA was detected by RT-PCR; the expression of p16 protein was detected by immunocytochemistry and Western blot. Results Homozygous Deletion of p16 was found in 3/6 pancreatic adenocarcinoma cell lines; expression of p16 was not detectable in these 3 cell lines in both mRNA and protein levels. Gene mutation and hy-permethylation of the promoter region of p16 were not found in all 6 cell lines. Conclusions Tumor suppressor gene p16 is inactivated by homozygous deletion in pancreatic adenocarcinoma cell lines.更多还原