乌头碱下调miR-23a诱导凋亡抑制胃腺癌细胞系MGC803增殖

Aconitine Inducesapoptosisand Inhibits Proliferation of Gastric Adenocarcinoma Cell Line MGC803 by down-regulating miR-23a

目的:研究乌头碱抑制人胃腺癌细胞系MGC803增殖的miRNA水平作用机制。方法:乌头碱作用于MGC803细胞后,MTT实验检测细胞活力并确定后续实验中乌头碱适宜的作用浓度,集落形成实验检测细胞增殖能力,TUNEL检测细胞凋亡情况,Real-Time PCR检测miR-23a的表达水平,Western blot检测miR-23a下游靶蛋白IRF1的表达水平。结果:乌头碱可降低MGC803细胞的存活率,且呈浓度及时间依赖性,同时乌头碱可降低肿瘤细胞的克隆形成能力;乌头碱可诱导细胞凋亡;乌头碱可下调miR-23a的表达,上调其下游靶蛋白IRF1的表达;封闭内源性miR-23a后经乌头碱处理,细胞凋亡水平显著增加。结论:乌头碱可通过调节miR-23a/IRF1通路,从而诱导胃腺癌细胞凋亡抑制细胞增殖。

Objective To explore the miRNAs related mechanism of aconitine in inhibiting proliferation of human gastric adenocarcinoma cells MGC803.Methods MGC803 cells treated with aconitine,MTT assay was used to detect cell viability and to determine the suitable concentration of aconitine for follow-up experiments,colony formation assay was used to detect ability ofcell clonogenicity,TUNEL was used to detect numbers of apoptotic cells,Real-time PCR was used to detect expression of miR-23a,Western blot was used to detect the expression of IRF1 targeted by miR-23a.Results Compared with the NC group,the cell viabilitywas significantly down-regulated,which exhibit dose-effect and time-effect relationship,and cell clonogenicity ability was also down-regulated;numbers of apoptotic cells was significantly up-regulated;the expression level of miR-23a was down-regulated,but the expression level of IRF1 protein was up-regulated in the aconitine group;Compared with NC group,MGC803 cells transfected with ASO-23a and then treated with aconitine was significantly up-regulated numbers of apoptotic cells.Conclusion Aconitine induces apoptosis and inhibits the proliferation of human gastric adenocarcinoma cells by regulating the miR-23a/IRF1 pathway.