摘要
目的研究miR-218对新藤黄酸抑制人宫颈癌He La细胞增殖作用的影响及其可能的分子机制。方法人宫颈癌He La细胞转染过表达真核表达载体pmiR-218,real-time PCR检测He La细胞miR-218的表达。将He La细胞分为空白对照组、新藤黄酸组、pmiR-218组、质粒对照组、新藤黄酸联合pmiR-218组。MTT法检测各组细胞增殖;流式细胞仪检测各组细胞凋亡;Western blot检测Bcl-2、Bax、E-cadherin表达;real-time PCR检测Bcl-2、Bax和E-cadherin的mRNA表达水平。结果 miR-218真核表达载体pmiR-218转染He La细胞后,细胞内miR-218表达水平明显增加(P<0.05)。miR-218可提高He La细胞对新藤黄酸的敏感性,高表达miR-218能促进新藤黄酸对He La细胞的增殖抑制作用,促进细胞凋亡,下调新藤黄酸对He La细胞Bcl-2/Bax蛋白的表达水平。结论 miR-218可提高He La细胞对新藤黄酸的敏感性,miR-218能增强新藤黄酸对He La细胞增殖抑制作用,并促进He La细胞凋亡,其作用机制可能与下调Bcl-2/Bax的表达有关。
Aim To study the inhibitory effects of gambogenic acid in combination with miR-218 on cervical cancer He La cells and its mechanisms. Methods Eukaryotic expression vector of miR-218（ pmi8-218） was transfected into He La cells. Transcript levels of miR-218 were quantified by real-time quantitative PCR.He La cells were incubated with different concentrations of gambogenic acid alone or in combination with pmiR-218. The cell growth inhibiting ratio of He La cells was assessed by MTT assay. Cell apoptosis was measured by fluorescence activated cell sorting. The expression levels of Bcl-2,Bax and E-cadherin were measured by Western blot and qRT-PCR. Results Levels of miR-218 transcript significantly increased in pmiR-218 transfected He La cells. Overexpression of miR-218 may enhance the sensitivity of He La cells to gambogenic acid. Over expression of miR-218 could enhance the effect of gambogenic acid on inhibition cell proliferation,promoting apoptosis of He La cells. pmiR-218 could enhance the regulation of Bax expression and decrease the expression of Bcl-2 in He La cells. Conclusions Over expression of miR-218 may enhance the sensitivity of He La cells to gambogenic acid; miR-218 can enhance the effect of gambogenic acid on inhibition cell proliferation and promote the apoptosis of He La cells,and the mechanism may be related to down-regulation of Bcl-2/Bax expression.
出处
《中国药理学通报》
CAS
CSCD
北大核心
2017年第10期1405-1409,共5页
Chinese Pharmacological Bulletin
基金
安徽省教育厅自然科学基金资助项目(No KJ2014A162)
