论文部分内容阅读
目的:建立Grim-19基因稳定表达的人子宫颈癌Hela细胞株,探讨Grim-19及其对下游基因STAT3表达的影响。方法:将构建好的质粒采用脂质体法转染人子宫颈癌Hela细胞株,经G418筛选,得到稳定表达Grim-19的Hela细胞株,并分别通过逆转录聚合酶链反应(RT-PCR)和免疫印迹法(Western-blot)检测Grim-19及其下游基因STAT3的表达。结果:成功建立了高表达GRIM-19的Hela/Grim-19细胞株,并在mRNA与蛋白水平上均发现Grim-19在转染细胞中的高表达和其下游基因STAT3表达的明显下调。结论:上调子宫颈癌Hela细胞Grim-19的表达对STAT3的表达有抑制作用。
OBJECTIVE: To establish a human cervical cancer Hela cell line with stable expression of Grim-19 gene and to investigate the effect of Grim-19 and its effect on STAT3 expression. Methods: The constructed plasmids were transfected into human cervical cancer Hela cells by lipofectamine. The cells were screened by G418 to obtain Hela cells stably expressing Grim-19. The recombinant plasmids were transfected into Hela cells by reverse transcription-polymerase chain reaction ) And Western-blot were used to detect the expression of Grim-19 and its downstream gene STAT3. Results: The Hela / Grim-19 cell line with high expression of GRIM-19 was successfully established. The high expression of Grim-19 in transfected cells and the down-regulation of STAT3 expression at mRNA and protein levels were found. Conclusion: The up-regulation of Grim-19 expression in cervical cancer Hela cells can inhibit the expression of STAT3.