骨髓基质细胞介导的微环境对人肝癌细胞株SMMC-7721的作用以及机制研究  

Effects of marrow stromal cell-mediated microenvironment on human hepatocellular carcinoma cell line SMMC-7721

作　　者：白慧丽[1] 李宝林[1] 何方[1] 张汝益[1] 严树涓[1] 刘晨[1] 杨丹丹[1] 施琼[1] 

机构地区：[1]重庆医科大学检验医学院临床检验诊断学教育部重点实验室,重庆400016

出　　处：《中国病理生理杂志》2013年第11期1921-1927,共7页Chinese Journal of Pathophysiology

基　　金：国家自然科学基金资助项目(No.NSFC 31200971);重庆市科委自然科学基金资助项目(No.CSTC;2011BB5131);重庆市教育委员会科学技术研究项目(No.KJ120327)

摘　　要：目的:探讨肿瘤微环境中人骨髓基质细胞HS-5对人肝癌细胞SMMC-7721的作用及机制。方法:采用HS-5细胞条件培养基(HS-5 cell-conditioned medium,HS-5-CM)处理肝癌细胞SMMC-7721,分别通过MTT、划痕和Transwell实验分析对SMMC-7721细胞增殖、迁移和侵袭的影响;采用Transwell小室间接共培养SMMC-7721细胞与HS-5细胞,应用实时荧光定量RT-PCR(qRT-PCR)、ELISA或Western blotting分别测定SMMC-7721细胞中趋化因子CCL5及其受体CCR5的mRNA和蛋白表达;此外加入PI3K抑制剂LY294002后,通过Western blotting检测SMMC-7721细胞中PI3K-Akt通路的相关蛋白Akt及其磷酸化水平的变化,以及qRT-PCR和ELISA检测SMMC-7721细胞中CCL5的变化。结果:HS-5-CM可以促进SMMC-7721细胞增殖、迁移和侵袭;间接共培养后,SMMC-7721细胞中CCL5及CCR5 mRNA和蛋白表达均有增高;PI3K抑制剂LY294002可有效抑制共培养后SMMC-7721细胞中PI3K-Akt通路的激活,并减少其CCL5的分泌。结论:HS-5细胞可以促进SMMC-7721细胞的增殖、迁移和侵袭能力;与HS-5细胞间接共培养,激活了肝癌细胞SMMC-7721的PI3K-Akt通路,从而增加SMMC-7721细胞的CCL5分泌。AIM： To investigate the effects of marrow stromal HS-5 cells on hepatocellular carcinoma SMMC-7721 cells in the tumor microenvironment. METHODS： The effects of HS-5 cell-conditioned medium （HS-5-CM） on the proliferation, migration and invasion abilities of SMMC-7721 cells were detected by MTY, wound-healing and Transwell assays. After co-culture of SMMC-7721 cells with HS-5 cells in the Transwell chamber, the expression of chemokine CCL5 and its receptor CCR5 at mRNA and protein levels in SMMC-7721 cells was examined by quantitative real-time PCR （ qRT- PCR）, ELISA or Western blotting. Akt and p-Akt473 protein levels in SMMC-7721 ceils treated with PI3K inhibitor LY294002 were observed by Western blotting. RESULTS： HS-5-CM promoted the proliferation, migration and invasion abilities of SMMC-7721 cells. The expression of CCL5 and CCR5 at mRNA and protein levels in SMMC-7721 cells was increased after co-cultured with HS-5 ceils. PI3K inhibitor LY294002 inhibited the activation of PI3K-Akt signaling pathway and the secretion of CCL5 in SMMC-7721 cells after co-cultured with HS-5 cells. CONCLUSION： HS-5 cells significantly promote the proliferation, migration and invasion abilities of SMMC-7721 cells. Co-culture of SMMC-7721 cells with HS-5 cells activates PI3K-Akt signaling pathway to increase the secretion of CCL5 in SMMC-7721 cells.

关 键 词：骨髓基质细胞 SMMC-7721细胞 趋化因子CCL5 微环境 

分 类 号：R735.7[医药卫生—肿瘤]