TGF-β上调lncRNA-ATB促进骨肉瘤细胞MG-63细胞转移的实验研究  

TGF-β Promotes Osteosarcoma Cell Mg-63 Cell Metastasis by Upregulating the Long Noncoding Rna-Atb

作　　者：吴玉娟 曹镐禄[1] 何华宾[2] 罗煜 温军 郭哲 余常辉[4] 吴冰 WU Yu-juan;CAO Gao-lu;HE Hua-bin;LUO Yu;WEN Jun;GUO Zhe;YU Chang-hui;WU Bing(School of Basic Medicine,Gannan Medical University,Ganzhou,Jiangxi 341000;The First Affiliated Hospital,Gannan Medical University,Ganzhou,Jiangxi 341000;Undergraduate Student in Grade 2013,Gannan Medical University,Ganzhou,Jiangxi 341000;Dept.of Respiratory Medicine,Nanfang Hospital,Southern Medical University,Guangzhou,Guangdong 510515)

机构地区：[1]赣南医学院基础医学院,江西赣州341000 [2]赣南医学院第一附属医院,江西赣州341000 [3]赣南医学院,江西赣州341000 [4]南方医科大学南方医院呼吸内科,广东广州510515

出　　处：《赣南医学院学报》2018年第1期5-10,共6页JOURNAL OF GANNAN MEDICAL UNIVERSITY

基　　金：国家自然科学基金项目(81560039);赣南医学院校级课题(YB201310)

摘　　要：目的:探讨lncRNA-ATB在TGF-β促进骨肉瘤细胞转移中的作用和机制。方法:用脂质体法将pro-ATB和shRNA-ATB的质粒转染到人骨肉瘤细胞MG-63细胞,用嘌呤霉素筛选稳定细胞株;用脂质体法将过表达mimics-30a或inhibitor-30a转染到MG-63细胞;Q-PCR法检测lncRNA-ATB和miR-30家族表达;Transwell法检测细胞转移能力;western blot检测ZEB1和E-cadherin蛋白表达。结果:TGF-β促进lncRNA-ATB和ZEB1蛋白表达,抑制miR-30a家族和E-cadherin蛋白表达,增加MG-63细胞转移能力;而敲低lncRNA-ATB表达后TGF-β的作用明显减弱。过表达lncRNA-ATB的MG-63细胞转移能力明显增强,ZEB1蛋白表达增加,miR-30a和E-cadherin蛋白表达下降;而mimics-30a可以抑制lncRNA-ATB的作用。相反,敲低lncRNA-ATB后,MG-63细胞转移能力和ZEB1蛋白表达明显减弱,miR-30a和E-cadherin蛋白表达增加,而inhibitor-30a可以逆转敲低lncRNA-ATB对MG-63细胞的影响。结论:TGF-β通过上调lncRNA-ATB抑制miR-30a表达,进而增加ZEB1表达及下调E-cadherin蛋白,从而促进骨肉瘤细胞转移。Objective:To investigate the effect and mechanism of Long Noncoding RNA-ATB(lncRNA-ATB)in promoting the metastasis of osteosarcoma cells induced by TGF-β.Methods:The plasmids of pro-ATB and shRNA-ATB were transfected into human osteosarcoma cell MG-63 by liposome method,and the stable cell lines were screened by purine mycin.Overexpression of mimics-30a or inhibitor-30a was transfected into MG-63 cells by liposome method.Q-PCR was used to detect lncRNA-ATB and miR-30 family expression.The metastasis ability of MG-63 cells was estimated by Transwell experiment.The expression of ZEB1 and E-cadherin proteins were detected by western blot.Results:TGF-βpromoted the expression of lncRNA-ATB and ZEB1 protein,and the metastasis ability of MG-63,but downregulated the expression of miR-30 family and E-cadherin protein.When the expression of lncRNA-ATB was downregulated,the effects of TGF-βwere obviously suppressed.Overexpression of lncRNA-ATB in MG-63 cells significantly enhanced their ability to metastasizeThe metastasis ability of MG-63 cells with overexpression of lncRNA-ATB enhanced significantly;the expression of ZEB1 protein increased;the expression of miR-30 family and E-cadherin protein were significantly reduced.But the mimics-30a could inhibit the effect of lncRNA-ATB.On the contrary,the metastasis ability and ZEB1 protein expression of MG-63 cells were significantly decreased,and the expression of miR-30 family and E-cadherin protein were significantly increased after lncRNA-ATB downregulated.But the inhibitor-30a could revere the effects of lncRNA-ATB downregulated on MG-63 cells.Conclusion:TGF-βinhibits the expression of miR-30a by upregulating lncRNA-ATB,thereby increasing the expression of ZEB1 and down-regulating E-cadherin protein,thus promoting the metastasis of osteosarcoma cells.

关 键 词：骨肉瘤细胞转移 lncRNA-ATB miR-30a TGF-Β 

分 类 号：R783.1[医药卫生—口腔医学]