靶向COX_医学论文.docVIP

靶向COX_医学论文 靶向COX_医学论文 作者：谯敏，向廷秀，王丕龙，黄爱龙 【关键词】 RNA干扰；环氧化酶 【Abstract】AIM: To investigate the inhibitory effect of siRNA targeting COX2 on the growth of gastric carcinoma cells of well, moderate or poor differentiation (MKN28, SGC7901, BGC823) and related mechanism. METHODS: siRNA targeting COX2 gene was designed, siRNACOX2 vector was constructed and transfected into gastric carcinoma cells to induce RNA interference. The changes of COX2 were detected with RTPCR and Western blotting.Cell viability was detected by MTT. Cell apoptosis was judged by TUNEL and electromicroscopy, and expressions of Bax and Bcl2 were tested by Western blot. RESULTS: Both COX2 expression and cell growth were inhibited in SGC7901 and BGC823 after transfection of siRNACOX2 vector into the cells. But, in MKN28 ,only COX2 expression decreased. The obvious cell apoptosis both in SGC7901 and BGC823 was observed. Bax was upregulated and Bcl2 was downregulated in SGC7901 and BGC823. In contrast, there were almost no changes in control group in vitro. CONCLUSION: RNA interference inhibits the growth of gastric carcinoma cells, which may be related to downregulation of COX2 and induction of cell apoptosis. The inhibitory effect of RNAi was relatively strong in SGC7901, which indicated that the effect of siRNA was dependent on the differentiation degree of gastric carcinoma cells. 【Keywords】 RNA interference COX2 cell proliferation apoptosis stomach neoplasms 【摘要】目的： 利用RNA干扰（RNA interference,RNAi）技术在体外抑制COX2表达，研究其抑制三种不同分化程度的胃癌细胞（MKN28，SGC7901，BGC823）的可能性，探讨RNAi抑制胃癌生长与胃癌细胞分化程度的相关性. 方法： 设计靶向COX2基因的siRNA，构建重组表达质粒pTZU6+1siRNACOX2并导入胃癌细胞株，体外诱导RNAi，采用逆转录PCR（RTPCR）和Western blotting检测COX2基因表达变化,MTT法检测细胞活力,原位末端标记（TUNEL）及透射电镜检测细胞凋亡，Western blotting检测凋亡相关蛋白Bax和Bcl2的改变. 结果： pTZU6+1siRNACOX2质粒导入细胞后，SGC7901和BGC823中COX2表达明显下调，细胞生长被抑制，并出现特征性的细胞凋亡，同时有Bax蛋白上调和Bcl2蛋白下调. 对照组则无明显变化. 结论： RNAi显著抑制胃癌细胞生长，可能与下调COX2基因表达和诱导细胞凋亡有关,其对中分化胃癌细胞的抑制作用最为显著，提示RNAi的作用可能依赖于胃癌细胞的分化程度. 【关键词】 RNA干扰；环氧化酶2；细胞增殖；细胞凋亡；胃肿瘤 环氧化酶（Cyclooxygenase