动力相关蛋白-1(MRP-1/CD9)对卵巢癌细胞体外增殖和运动的影响

The Effect of MRP-1/CD9 on Cell Proliferation and Motility of Human Ovarian Cancer Cell Lines in Vitro

目的:动力相关蛋白-1(MRP-1/CD9)是四跨膜蛋白超家族(transmembrane4,superfamily,TM4SF)成员之一,参与调控细胞的生长、分化、细胞间粘附和迁移。本实验旨在研究MRP-1/CD9对人卵巢癌SKOV-3细胞株体外增殖和运动能力的影响并探讨其与PI4K/AKT信号通路的相关性。方法:应用RT-PCR获得CD9全长cDNA片段,正向插入pcDNA3.1表达载体,将重组质粒导入SKOV-3细胞中;应用RT-PCR、cfse-流式细胞测定和单层伤口愈合实验等方法观察转染前后SKOV-3细胞PI4K和AKTmRNA表达水平,细胞增殖及其运动能力变化。结果:成功构建正义全长CD9真核表达载体,获得稳定表达CD9的SKOV-3克隆株。与空质粒转染和不转染的SKOV-3细胞相比,SKOV-3/CD9细胞PI4KmRNA的表达明显被抑制,抑制率为40%;AKTmRNA的表达水平上调3.13倍,细胞增殖和运动能力均有明显升高。结论:CD9促进人卵巢癌SKOV-3细胞的体外增殖和运动能力,可能是通过PI4K/AKT信号通路发挥作用的,对卵巢癌恶性进展发挥重要作用。

Objective：To investigate the effects of transfecting the MRP-1/CD9 gene into the human ovarian careinoma cell line SKOV-3 on tumor cell proliferation and motility and to observe the relationship between CD9 and the PI4K/AKT signaling pathway.Methods：Full-length cDNA was obtained for CD9 using RT-PCR and was inserted into the peDNA3.1 expression veetor,The reconbinant plasmid was introduced into the SKOV-3cells RT-PCR,CFSE-flow cytometry and moostratal wound healing were used to detemine the expression levels of PI4K and AKT.cell proliferation and mtility of the SKOV-3cells,respeetively,before and after the transfection,Resultss：The eukaryotic expression vector was constructed successfully and clones with stable overexpression of CD9 were obtained,Compared with mock plasmid-transfeeted and untransfected cells,the mRNA expression of the PI4K gene was signifcantly inhibited by 40% and AKT mRNA expression was upregulated 3.13-fold in cells containing the CD9expression vector.Both tumor cell proliferation and cell motility were notably enhanced in SKOV-3/CD9 cell clones.Conclusion：CD9 overexpression increased cell proliferation and motility of SKOV-3 cells,and these functions may be induced by activating the PI4K/AKT signaling pathway. This study suggests overexpression of CD9 is associated with a more malignant progression in ovarian carcinoma.