基础医学与临床 ›› 2016, Vol. 36 ›› Issue (8): 1118-1123.

• 研究论文 • 上一篇    下一篇

BMP4沉默对骨肉瘤细胞间系MG-63上皮间质转换及侵袭转移能力的影响

王顺义1,李辉2,任东良1,马伟松1,李伟峰1,史占雷2,刘向东1   

  1. 1. 保定市第一中心医院
    2. 河北省保定市第一中心医院
  • 收稿日期:2015-12-07 修回日期:2016-03-26 出版日期:2016-08-05 发布日期:2016-07-13
  • 通讯作者: 王顺义 E-mail:wangshunyi_doctor@126.com

Effects of silencing BMP4 on mesenchymal epithelial transition, invasion and migration ability of osteosarcoma cell line MG-63

  • Received:2015-12-07 Revised:2016-03-26 Online:2016-08-05 Published:2016-07-13

摘要: 目的 探讨沉默骨形成蛋白-4(BMP4)对骨肉瘤细胞系MG-63上皮间质转化转换(EMT)和侵袭迁移能力的影响。方法 脂质体法转染MG-63细胞BMP4 shRNA表达质粒,将MG-63细胞分为MG-63对照组(正常培养对照)、空白对照组(转染空白质粒)和BMP4沉默组(转染BMP4 shRNA质粒)。逆转反转录聚合酶链反应(RT-PCR)法和Western blot免疫印迹法检测细胞E-cadherin,、vimentin、twist和snail的mRNA及蛋白水平。Transwell侵袭实验和伤口愈合实验检测细胞侵袭和迁移能力。结果 沉默BMP4后MG-63细胞的间质细胞标志物vimentin、twist1和snail水平降低,上皮细胞标志物E-cadherin表达增高(P<0.05)。BMP4沉默组细胞穿膜细胞数和迁移距离显著少于MG-63对照组和空白对照组(P<0.05)。结论 沉默BMP4基因可以使间质型MG-63骨肉瘤细胞向上皮细胞表型转变,从而有效地降低肿瘤细胞的侵袭迁移能力。

关键词: 骨肉瘤, 骨形成蛋白-4, 上皮间质转化, 侵袭, 迁移

Abstract: Objective To investigate the effects of silencing BMP4 on mesenchymal epithelial transition, invasion and migration ability of osteosarcoma cell line MG-63. Methods BMP4 shRNA plasmid was transfected into MG-63 cells,MG-63 cells were divided into MG-63 control group(normal culture control),empty control group(transfected empty plasmid),MG-63silence group(transfected BMP4 silence shRNA). Reverse transcription polymerase chain reaction (RT-PCR) and Western blotting were used to detect the changes of E-cadherin,vimentin、twist and snail mRNA and protein levels after the cells BMP4 was silenced. Transwell invasion assay and wound healing assay were conducted when the BPM4 was silenced to detect the changes of the invasion and migration ability of MG-63 cells. Results Silencing BMP4 decreased MG-63 cells twist and snail expression, increased the expression of epithelial cell marker E-cadherin, and decreased expression of vimentin. Invasion and migration assays showed that the transmembrane cell number and migration distance of BMP4 silence group were significantly less than those of the MG-63 control group and empty control group(P<0.05).Conclusions The silence of BMP4 gene may change MG-63 osteosarcoma cells from mesenchymal phenotype to epithelial phenotype, reducing tumor cell invasion and migration ability effectively.

Key words: osteosarcoma, BMP4, epithelial mesenchymal transition, invasion, migration

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