δ-三烯生育酚通过上调MicroRNA-34a抑制宫颈癌细胞增殖

δ-tocotrienol Inhibits Proliferation of Cervical Carcinoma Cells Through Upregulating MicroRNA-34a

目的探究δ-三烯生育酚对HeLa宫颈癌细胞增殖的影响以及microRNA-34a(miR-34a)是否介导这一过程。方法用荧光定量PCR方法检测miR-34a和细胞增殖关键基因表达的变化,CCK-8检测细胞活力变化,Western blot检测细胞增殖关键基因蛋白水平的变化,流式细胞术检测细胞周期变化;并利用miR-34a模拟剂和抑制剂,探究其对细胞增殖的影响。结果δ-三烯生育酚处理HeLa宫颈癌细胞36 h显著降低细胞活力并呈剂量依赖效应。用20μmol/Lδ-三烯生育酚处理细胞36 h显著上调miR-34a表达,显著下调细胞周期关键基因CCND1,同时S细胞比例显著下降。细胞转染miR-34a模拟剂后,CCND1表达和细胞活力都下降;而转染miR-34a抑制剂则显著减弱δ-三烯生育酚的增殖抑制作用。结论δ-三烯生育酚通过上调miR-34a抑制宫颈癌细胞增殖。

Objective To determine the effects ofδ-tocotrienol on the proliferation of cervical carcinoma HeLa cells and whether miR-34a is involved in this process.Methods The expression of miR-34a and proliferation-related genes,cell viability,the protein levels of proliferation-related genes and cell cycle were examined using Real-time PCR,CCK-8 assay,Western blot and flow cytometry,respectively.miR-34a mimics and inhibitors were used to determine whether miR-34a mediated the effects ofδ-tocotrienol on cell proliferation.Resultsδ-tocotrienol treatment for 36h significantly decreased the viability of HeLa cells in a dose-dependent manner.δ-tocotrienol treatment at 20μmol/L-1 for 36h significantly stimulated miR-34a expression,however,inhibited CCND1 expression;meanwhile,cells proportion in S phase was decreased.Besides,after miR-34a mimics transfection,the cell viability and CCND1 expression were also decreased.Furthermore,miR-34a inhibitor transfection attenuated the inhibitory effects ofδ-tocotrienol on cell proliferation.Conclusionδ-tocotrienol inhibits the proliferation of cervical carcinoma cells through upregulating miR-34a expression.