miR-505对骨肉瘤细胞增殖、侵袭的影响及相关机制研究  

Effect of miR-505 on proliferation and metastasis of osteosarcoma cells and its related mechanisms

作　　者：杨晋[1] 瓦庆德[1] 余浩[1] 杨继滨 桑鹏[1] Yang Jin;Wa Qingde;Yu Hao;Yang Jibin;Sang Peng(Department of Orthopedics,Affiliated Hospital of Zunyi Medical College)

机构地区：[1]遵义医学院附属医院骨一科,遵义563003

出　　处：《重庆医科大学学报》2020年第2期228-233,共6页Journal of Chongqing Medical University

基　　金：贵州省科技合作计划资助项目(编号:黔科合LH〔2015〕7496)。

摘　　要：目的:探讨miR-505对骨肉瘤细胞增殖、侵袭的影响及相关机制。方法:采用q-PCR测定人成骨细胞hFOB1.19及骨肉瘤细胞MG63中的miR-505水平。分别设置miR-505 mimics组与阴性对照(miR-NC)组,采用Lipofectamine TM 2000将miR-505 mimics及miR-NC转染至MG63细胞中,比较2组细胞增殖速率及侵袭细胞数,进一步采用Western blot及q-PCR考察转染mi R-505的MG63细胞中MMP-9、CDK4、Cyclin-D1、HMGB-1及β-联蛋白(β-catenin)、磷酸糖原合成酶激酶-3β(GSK-3β)的蛋白及mRNA水平。结果:骨肉瘤细胞MG63中miR-505水平为0.57±0.06,明显低于人成骨细胞hFOB1.19的1.03±0.12(P<0.01)。转染miR-505后,miR-505 mimics组的表达量为3.59±0.37,明显高于miR-NC组的1.09±0.12(P<0.01)。miR-505 mimics组转染48 h和72 h后的OD450 nm值分别为0.84±0.09和1.29±0.13,均明显低于miR-NC组(P<0.05)。miR-505mimics组侵袭细胞数为(42.84±4.35)个,明显低于miR-NC组的(66.84±6.85)个(P<0.05)。miR-505 mimics组MMP-9、CDK4、Cyclin-D1及HMGB-1蛋白水平均低于miR-NC组(P<0.05)。miR-505 mimics组MMP-9、CDK4、Cyclin-D1及HMGB-1 mRNA水平均低于miR-NC组(P<0.05)。同时,miR-505 mimics组Wnt信号通路中关键靶点β-catenin和GSK-3β蛋白与基因表达均明显低于miR-NC组(P<0.05)。结论:miR-505可通过抑制Wnt信号通路,下调MMP-9、CDK4、Cyclin-D1及HMGB-1表达,从而抑制骨肉瘤细胞的增殖与侵袭。Objective:To investigate the effect of miR-505 on proliferation and metastasis of osteosarcoma cells and its mechanisms.Methods:The miR-505 level in human osteoblast of hFOB1.19 and osteosarcoma of cell MG63 was determined by q-PCR. The miR-505 mimics group and the negative control group(miR-NC) were set. Both miR-505 mimics and miR-NC were transfected into MG63 cells with Lipofectamine TM 2000 and cell proliferation rate and invasive cells were compared between two groups. Proteins and mRNA levels of MMP-9,CDK4,Cyclin-D1,HMGB-1,β-catenin and GSK-3β in MG63 cells transfected with miR-505 was further detected via Western blot and q-PCR technologies. Results:The expression level of miR-505 in osteosarcoma cell of MG63 was 0.57±0.06,which was significantly lower than that of 1.03±0.12 in human osteoblast of hFOB1.19(P<0.01). After transfecting miR-505,expression level in the miR-505 mimics group was 3.59±0.37,which was significantly higher than that of 1.09±0.12 in the miR-NC group(P<0.01). After transfecting 48 h and 72 h,A450 nm values in the miR-505 mimics group were 0.84±0.09 and1.29±0.13,respectively,which were significantly lower than those in the miR-NC group(P<0.05). Invasive cells in the miR-505 mimics group was(42.84±4.35),which was significantly lower than that of(66.84±6.85) in miR-NC group(P<0.05). Protein levels of MMP-9,CDK4,Cyclin-D1 and HMGB-1 in the miR-505 mimics group were lower than those in the miR-NC group(P<0.05).The mRNA levels of MMP-9,CDK4,Cyclin-D1 and HMGB-1 in the miR-505 mimics group were lower than those in the miR-NC group(P<0.05). Meanwhile,protein and gene expressions of β-catenin and GSK-3β in Wnt signaling pathway in the miR-505 mimics group were significantly lower than those in the miR-NC group(P<0.05). Conclusion:The miR-505 can down-regulate the expression of MMP-9,CDK4,Cyclin-D1 and HMGB-1 by inhibiting Wnt signaling pathway,so as to inhibit the proliferation and metastasis of osteosarcoma cells.

关 键 词：miR-505 骨肉瘤 细胞增殖 细胞侵袭 WNT信号通路 

分 类 号：R738.1[医药卫生—肿瘤]