BMP4沉默对骨肉瘤细胞系MG-63上皮间质转换及侵袭转移能力的影响

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

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)。反转录聚合酶链反应(RT-PCR)法和Western blot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。

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 poly- merase chain reaction （RT-PCR） and Western blot were used to detect the changes of E-cadherin ,vimentin, twist and snail mRNA and assay were conducted MG-63 ceils. Results protein levels after the cells BMP4 was silenced. Transwell invasion assay and wound healing when the BPM4 was silenced to detect the changes of the invasion and migration ability of 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 ceils from mesenchymal phenotype to epithelial phenotype, reducing tumor cellinvasion and migration ability effectively.