槲皮素通过提高miR-199a表达抑制肝癌的实验研究
Effects of quercetin on proliferation of hepatocarcinoma cell line HepG2 by up-regulating miR-199a expression
张味娜 1张明明 1俞敏1
作者信息
- 1. 宁海县第一医院 药剂科,浙江 宁波 315600
- 折叠
摘要
目的 探究槲皮素通过提高miR-199a表达抑制肝癌的实验研究.方法 用20μm/L槲皮素处理HepG2肝癌细胞,MTT法比较不同处理时间(0、12、24、48 h)槲皮素组和对照组的细胞增殖情况,流式细胞术检测各组细胞凋亡情况;同时RT-qPCR检测miR-199a的表达,Western blotting检测各组凋亡相关基因VEGFA、Bax、Caspase3的表达情况.结果 MTT结果显示槲皮素具有抑制肝癌细胞的增殖能力(P<0.01),流式细胞术结果显示,槲皮素处理后肝癌细胞凋亡增加(P<0.01).并且槲皮素处理能够增加肝癌细胞中miR-199a的表达,抑制VEGFA表达(P<0.01),同时促进凋亡基因Bax和Caspase3的mRNA和蛋白水平的表达(P<0.01).结论 槲皮素能够通过上调肝癌细胞中miR-199a的表达,抑制肝癌细胞的增殖并促进其凋亡.
Abstract
Objective To study the effect of quercetin on proliferation of hepatocarcinoma cells and the expression of miR-199a. Methods Hepatocellular carcinoma cell line HepG2 cells were transfected with 20 μm/L quercetin. MTT proliferation assay were conducted to compare the proliferation, and Annexin V/PI apoptosis assay were adopted to compare the apoptosis of cells in quercetin group and NC group at 12 h, 24 h, 36 h and 48 h. The expression of miR-199a were determined by RT-qPCR and western blotting assays were used to detect the Western blotting was used to detect the expression of VEGFA, Bax and Caspase3. Results MTT showed that quercetin inhibited the proliferation of HepG2 cells (P<0.01). The apoptosis of hepatocellular carcinoma was markedly increased after treated with quercetin (P<0.01). Moreover, quercetin can up-regulate the expression of miR-199a in HepG2 cells. The quercetin decreased the expression of VEGFA, increased the expression of Bax and Caspase 3 on both mRNA and protein level (P<0.01). Conclusion Quercetin can inhibit the proliferation and promote apoptosis of hepatocarcinoma cells by up-regulating the expression of miR-199a.
关键词
槲皮素/肝癌HepG2细胞/增殖/凋亡/miR-199aKey words
quercetin/HepG2/proliferation/apoptosis/miR-199a引用本文复制引用
基金项目
浙江省药学会专项科研资助项目(2014ZYY38)
出版年
2017
NETL
NSTL
维普
万方数据