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

目的：动力相关蛋白-1（MRP-1／CD9）是四跨膜蛋白超家庭（transmetubrane 4,stperfanily,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SKON-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 efse一流式细胞测定和单层伤口愈合实验等方法观察梁前后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,与穿空质粒转染和不转染的SKON-3细胞相比,SKON-3/CD9细胞PI4KmRNA的表达明显被抑制,抵制率为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.

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

Objective:To investigate the effects of transfecting the MRP-1/CD9 gene into the human ovarian carcinoma 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 pcDNA3.1 expression vector.The recombinant plasmid was introduced into the SKOV-3 cells.RT-PCR,CFSE-flow cytometry and monostratal wound healing were used to determine the expression levels of PI4K and AKT,cell proliferation and motility of the SKOV-3 cells,respectively,before and after the transfection.Results:The eukaryotic expression vector was constructed successfully and clones with stable overexpression of CD9 were obtained.Compared with mock plasmid-transfected and untransfected cells,the mRNA expression of the PI4K gene was significantly inhibited by 40% and AKT mRNA expression was upregulated 3.13-fold in cells containing the CD9 expression 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.