下调miR-19b表达对骨肉瘤MG63细胞侵袭、迁移及Wnt/β-Catenin通路的影响  

Effects of Down-regulation of miR-19b Expression on the Invasion,Migration and Wnt/β-Catenin Pathway of Osteosarcoma MG63 Cells

作　　者：潘寒松 李祥[1] 郭昌军[1] 洪荣华[1] Pan Hansong;Li Xiang;Guo Changjun;Hong Ronghua(Department of Orthopaedics,Taizhou First People's Hospital,Zhejiang Taizhou 318020,China)

机构地区：[1]台州市第一人民医院骨科,浙江台州318020

出　　处：《中国药师》2020年第6期1032-1036,共5页China Pharmacist

基　　金：台州市社会发展类一般项目(编号:1701KY48)。

摘　　要：目的:探讨下调miR-19b表达对骨肉瘤MG63细胞侵袭、迁移能力的影响,并探讨可能机制。方法:采用脂质体转染法转染人骨肉瘤MG63细胞,实验分为空白组、miR-19b inhibitor NC组、miR-19b inhibitor组,采用细胞计数试剂盒-8(CCK-8)法检测细胞增殖情况,通过Transwell小室法检测各组细胞侵袭、迁移情况,通过免疫印迹(WB)法检测侵袭、迁移以及Wnt/β-连环蛋白(β-Catenin)通路相关蛋白表达情况。结果:与空白组、miR-19b inhibitor NC组相比,miR-19b inhibitor组miR-19b相对表达量、转染后48 h、72 h吸光度(A)值、侵袭与迁移细胞数、基质金属蛋白酶-2(MMP-2)、基质金属蛋白酶-9(MMP-9)、Wnt、细胞核β-Catenin表达水平显著降低(P<0.05),细胞质β-Catenin表达水平显著升高(P<0.05)。结论:下调miR-19b表达可能通过抑制Wnt/β-Catenin信号通路激活,从而抑制骨肉瘤MG63细胞侵袭、迁移。Objective:To investigate the effects of down-regulation of miR-19b expression on the invasion and migration of osteosarcoma MG63 cells,and to explore the possible mechanism.Methods:Human osteosarcoma MG63 cells were transfected with liposomes,and divided into the blank group,miR-19b inhibitor NC group and miR-19b inhibitor group.Cell counting kit-8(CCK-8)method was used to detect the cell proliferation,Transwell method was used to detect the cell invasion and migration,and Western blotting(WB)was used to detect the invasion,migration and Wnt/β-Catenin pathway-related protein expression.Results:Compared with that in the blank group and miR-19b inhibitor NC group,the relative expression of miR-19b in miR-19b inhibitor group decreased significantly(P<0.05).CCK8 test showed that compared with those in the blank group and miR-19b inhibitor NC group,A values of miR-19b inhibitor group decreased significantly at 48 h and 72 h after the transfection(P<0.05).Transwell test showed that the number of invasive and migrating cells in miR-19b inhibitor group was significantly lower than that in the blank group and miR-19b inhibitor NC group(P<0.05).WB results showed that compared with those in the blank group and miR-19b inhibitor NC group,the expression levels of matrix metalloproteinase-2(MMP-2),matrix metalloproteinase-9(MMP-9),Wnt and nuclearβ-Catenin in miR-19b inhibitor group decreased(P<0.05),while the expression level of cytoplasmicβ-Catenin increased(P<0.05).Conclusion:Down-regulation of miR-19b expression may inhibit the invasion and migration of osteosarcoma MG63 cells by inhibiting the activation of Wnt/β-Catenin signaling pathway.

关 键 词：骨肉瘤 MG63细胞 miR-19b 侵袭 迁移 Wnt/β-连环蛋白通路 

分 类 号：R965.1[医药卫生—药理学]