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目的:研究肿瘤生长抑制基因4(inhibitor of growth family4,ING4)体外对胃癌细胞SGC-7901的生长抑制及其分子机制。方法:将携带有人ING4基因的重组复制缺陷型腺病毒(Ad-hING4)感染胃癌细胞SGC-7901,RT-PCR法检测腺病毒介导的外源性hING4基因的转录,MTT法检测hING4基因对SGC-7901细胞的生长抑制作用,激光扫描共聚焦显微镜(LSCM)观察hING4基因诱导SGC-7901细胞凋亡的形态学改变,流式细胞仪(FACS)检测hING-4基因诱导胃癌细胞SGC-7901的凋亡率及细胞周期改变,免疫细胞化学分析凋亡相关因子bcl-2和bax的改变。结果:Ad-hING4感染SGC-7901细胞后,RT-PCR结果提示有hING4目的基因的转录,该基因表达可以显著抑制SGC-7901细胞的生长,诱导S期减少、G2/M期阻滞和凋亡,hING4基因能使SGC-7901细胞中bcl-2的表达下调和bax的表达上调。结论:重组腺病毒Ad-hING4对胃癌细胞SGC-7901具有显著的生长抑制及凋亡诱导作用,其作用机制可能与下调bcl-2基因和上调bax基因的表达有关。
Objective: To study the inhibitory effect of ING4 on the growth of gastric cancer cell line SGC-7901 and its molecular mechanism. Methods: Human gastric adenocarcinoma cell line SGC-7901 was infected by recombinant replication-defective adenovirus carrying human ING4 gene. The transcription of exogenous hING4 gene was detected by RT-PCR and hING4 gene by MTT assay SGC-7901 cells. The morphological changes of SGC-7901 cells induced by hING4 gene were observed by laser scanning confocal microscopy (LSCM). The apoptosis of SGC-7901 cells induced by hING-4 gene was detected by flow cytometry (FACS) The changes of apoptosis and cell cycle were analyzed by immunocytochemistry. The changes of apoptosis-related factors bcl-2 and bax were analyzed. Results: After transfected with Ad-hING4, the transcription of hING4 gene was detected by RT-PCR. The gene expression could significantly inhibit the growth of SGC-7901 cells, induce the decrease of S phase and arrest the G2 / M phase HING4 gene can down-regulate the expression of bcl-2 and bax in SGC-7901 cells. Conclusion: Recombinant adenovirus Ad-hING4 can significantly inhibit the growth of SGC-7901 cells and induce the apoptosis of gastric cancer cells. Its mechanism may be related to the down-regulation of bcl-2 gene and up-regulation of bax gene expression.