The Effect of MRP-1/CD9 on Cell Proliferation and Motility of Human Ovarian Cancer Cell Lines in Vitro
-
摘要: 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。Abstract: 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.
-
Keywords:
- Motility related protein-1 /
- Ovarian carcinoma /
- Cell proliferation /
- Cell motility
-
-
[1] 1 Maecker HT, Todd SC, Levy S. The tetraspanin superfamily: molecular facilitators [J]. FASEB J, 1997, 11(6):428~442
2 Funakoshi1 T, Tachibana I, Hoshida Y, et al. Expression of tetraspanins in human lung cancer cells:frequent downregulation of CD9 and its contribution to cell motility in small cell lung cancer [J]. Oncogene, 2003, 22(5):674~687
3 Hori H, Yano S, Koufuji K, et al. CD9 Expression in Gastric Cancer and Its Significance [J]. JSurg Res, 2004, 117(2):208~215
4 Sauer G, Windisch J, Kurzeder C, et al. Progression of Cervical Carcinomas Is Associated with Down- Regulation of CD9 But Strong Local Re- expression at Sites of Transendothelial Invasion [J]. Clin Cancer Res, 2003, 9(17):6426~6431
5 Zvieriev V, Wang JC, Chevrette M. Over- expression of CD9 does not affect in vivo tumorigenic or metastatic properties of human prostate cancer cells [J]. Biochem Biophys Res Commun, 2005, 337(2):498~504
6 Valster A, Tran NL, Nakda M, et al. Cell migration and invasion assays[J]. Methods, 2005, 37(2):208~215
7 Furuya M, Kato H, Nishimura N, et al. Down- regulation of CD9 in Human Ovarian Carcinoma Cell Might Contribute to Peritoneal Dissemination: Morphologic Alteration and Reduced Expression of B1 Integrin Subsets [J]. Cancer Res, 2005, 65 (7): 2617~2625
8 Gesierich S, Paret C, Hildebrand D, et al. Colocalization of the Tetraspanins, CO - 029 and CD151, with Integrins in Human Pancreatic Adenocarcinoma: Impact on Cell Motility [J]. Clin Cancer Res, 2005, 11(8):2840~2852
9 Hemler ME. Tetraspanin functions and associated microdomains [J]. Nat Rev Mol Cell Biol, 2005, 6(10):801~811
10 Larue L, Bellacosa A. Epithelial- mesenchymal transition in development and cancer: role of phosphatidylinositol 3' kinase/AKT pathways [J]. Oncogene, 2005, 24(50):7443~7454
计量
- 文章访问数: 6
- HTML全文浏览量: 0
- PDF下载量: 1
下载:
