摘要
目的:探讨白藜芦醇通过调控微小RNA-34a(miR-34a)表达对骨肉瘤MG-63细胞生长、侵袭和自噬的影响。方法:将骨肉瘤MG-63细胞分为对照组及10、20、40、60和80μmol/L白藜芦醇处理组。采用MTT法、Transwell小室和Western blot实验检测各组骨肉瘤细胞的活力、侵袭能力及自噬蛋白表达;RT-q PCR检测各组骨肉瘤细胞中miR-34a的表达。转染miR-34a模拟物(miR-34a mimic)及阴性对照(miR-34a NC)至骨肉瘤细胞,检测miR-34a mimic和miR-34a NC在白藜芦醇处理后对骨肉瘤细胞活力和侵袭能力的影响,Western blot检测自噬标志蛋白LC3-Ⅰ、LC3-Ⅱ和beclin-1的蛋白表达。结果:与对照组相比,不同浓度的白藜芦醇可抑制肿瘤细胞的活力和侵袭能力,促进自噬(P<0.05)。白藜芦醇可上调肿瘤细胞中miR-34a的表达,并具有剂量依赖性(P<0.05),同时促使LC3-Ⅱ/LC3-Ⅰ比值升高,beclin-1蛋白表达量上调(P<0.05)。miR-34a mimic可增加白藜芦醇对骨肉瘤细胞活力和侵袭能力的抑制,并进一步促进自噬。结论:白藜芦醇可能通过上调miR-34a表达抑制骨肉瘤MG-63细胞生长并促进其自噬。
AIM： To investigate the effects of resveratrol on the viability,invasion and autophagy of osteosarcoma MG-63 cells and the regulatory effect of microRNA-34a（ miR-34a）. METHODS： MG-63 cells were divided into 6 groups： control group and resveratrol treatment groups at doses of 10,20,40,60 and 80 μmol/L. MTT assay,Transwell chamber method and Western blot were used to detect the effects of resveratrol on the viability,invasion ability and expression of autophagy-related proteins in the osteosarcoma cells. The effect of resveratrol at different concentrations on the expression of miR-34a in the osteosarcoma cells was detected by RT-q PCR. The effects of miR-34a mimic and miR-34a mimic negative control（ miR-34a NC） transfection on the viability and invasion ability of osteosarcoma cells after treated with resveratrol at different concentrations were analyzed. The effects of miR-34a mimic transfection on autophagy-related proteins LC3-I,LC3-II and beclin-1 were determined by Western blot. RESULTS： Compared with control group,resveratrol inhibited the viability and invasion ability of the MG-63 cells and promoted autophagy（ P〈0. 05）. Resveratrol up-regulated the expression of miR-34a in the MG-63 cells in a dose-dependent manner（ P〈0. 05）. In addition,the ratio of LC3-II/LC3-I was increased,and beclin-1 was up-regulated（ P〈0. 05）. Co-treatment with miR-34a mimic and resveratrol increased inhibitory effects of resveratrol on the viability and invasion ability and invasion of the MG-63 cells and also promoted autophagy. CONCLUSION： Resveratrol inhibits the viability and invasion of osteosarcoma MG-63 cells and promotes autophagy by up-regulating miR-34a expression.
作者
直彦亮
张震
贺宪
ZHI Yan-liang;ZHANG Zhen;HE Xian(Panyu Hospital of Chinese Medicine,Guangzhou 511400,China;Shenzhen Baoan Shajing People＇s Hospital,Guangzhou Medical University,Shenzhen 518104,China)
出处
《中国病理生理杂志》
CAS
CSCD
北大核心
2018年第8期1455-1460,共6页
Chinese Journal of Pathophysiology
基金
广东省中医药局科研项目(No.20181217)
