摘要
目的:我们已经报道锰型超氧化物岐化酶(MnSOD)高表达可明显抑制SGC7901胃癌细胞的增殖,本研究拟进一步探索其中所涉及的分子机制.方法:利用DCFH-DA荧光染色方法检测正义、反义及空载SGC7901胃癌细胞株内的活性氧水平透射电镜下观察肿瘤细胞形态及细胞器结构改变.RT—PCR方法检测三者缺氧诱导因子HIF-1α的表达.结果:正义MnSOD-7901胃癌细胞胞内ROS下降,HIF-1α表达下调.反义组电镜下细胞形态明显小于正义组,正义组线粒体明显较反义组丰富,但两组线粒体均呈空泡状,缺少线粒体嵴.反义组核糖体明显较正义组丰富,内质网也略多.结论:MnSOD依赖的抑制胃癌细胞增殖途径与下调肿瘤的胞内ROS水平,下调HIF-1α的表达及改变细胞器的超微结构有关.
AIM: To investigate the molecular mechanisms involved in maganese superoxide dismutase (MnSOD) mediated growth inhibition of gastric cancer cell line SGC7901. METHODS: Sense and antisense MnSOD were stably tranfected into gastric cancer cell line SGC7901 to establish the sublines of MnSOD-7901 and MnSOD-AS7901. The levels of endogenous reactive oxygen species (ROS) in MnSOD-7901 and MnSOD-AS7901 cells were evaluated by DCFH-DA fluorescence staining assay, and then were compared with that in the mock transfectant (vector-7901). The ultrastructures of the cells were observed by transmission electron microscopy. The transcription of hypoxia in-ducible factor 1 α (HIF-1 α) in the cells was detected by reverse transcription polymerase chain reaction (RT-PCR). RESULTS: The level of ROS was decreased, and the ex- pression of HIF-1α was down-regulated in MnSOD-7901 cells as compared with that in vector-7901. MnSOD-AS7901 cells were much smaller in size and had few mitochondrions as compared with MnSOD-7901 cells, but vacuole-like mitochondrions and incomplete mitochondrial cristaes were observed in both kinds of cells. In addition, the endoplasmic reticula and ribosomes were also more plentiful in MnSOD-AS7901 cells. CONCLUSION: The MnSOD-dependent growth inhibition of gastric cancer cells is associated with the reduction of endogenous ROS and down-regulation of HIF-1α expression. In addition, the organelle alteration is also involved in this process.
出处
《世界华人消化杂志》
CAS
北大核心
2005年第12期1386-1389,共4页
World Chinese Journal of Digestology
基金
国家自然科学基金重点资助项目
No.30130100~~
