摘要
目的探讨下调miR-886-5p的表达对SiHa细胞增殖的影响。方法应用脂质体转染法分别将miR-886-5p表达抑制质粒及其对照质粒转染SiHa细胞,经Blasticidin抗生素筛选,得到稳定抑制miR-886-5p表达的SiHa细胞(SiHa miR-886-5p inhibitor)和稳定转染对照质粒的细胞;RT-PCR进行验证;采用MTT测增殖及克隆形成实验检测细胞的增殖能力。结果获得了稳定抑制miR-886-5p表达的宫颈癌SiHa细胞系;MTT测增殖实验发现,SiHa miR-886-5p inhibitor体外增殖能力比SiHa miR-886-5p NC及未转染质粒组细胞均明显减弱(P<0.05);克隆形成实验发现,SiHa miR-886-5p inhibitor体外克隆形成能力比SiHa miR-886-5p NC及未转染质粒组细胞均明显减弱(P<0.01)。结论下调miR-886-5p可显著抑制SiHa细胞的增殖,提示靶向抑制miR-886-5p为宫颈癌的治疗提供了新思路。
Objective To investigate the effects of miR- 886 - 5p knockdown on proliferation in SiHa cells. Methods SiHa cells were transfected with miR- 886 - 5p knockdown plasmid and negative control plasmid. After selected with Blasticidin, stable miR- 886 - 5p knockdown (SiHa miR- 886 - 5p inhibitor) and negative control clones (SiHa miR- 886 - 5p NC) were obtained, and then a qRT - PCR was used to examine the transfection efficiency of cells. Furthermore, MTT for proliferation and colony formation assay were used to examine the effect of miR- 886 - 5p to SiHa cells. Results The miR - 886 - 5p knockdown cell lines of cervical cancer were obtained, and results showed that proliferation ability of SiHa miR- 886 - 5p inhibitor was weaker than SiHa miR- 886 - 5p NC and SiHa untransfected by means of MTT methods (P〈0.05) and colony formation assay (P〈0.01). Conclusion The miR- 886 - 5p knockdown can signif- icantly suppress the celluar proliferation in SiHa cells. It provides a new idea for the treatment of cervical cancer by inhibiting miR - 886 - 5p.
出处
《中国煤炭工业医学杂志》
2014年第9期1489-1492,共4页
Chinese Journal of Coal Industry Medicine
基金
河北省秦皇岛市科技支撑计划项目(编号:2012023A114)