过表达CCR9对SW480结肠癌细胞侵袭的影响.docVIP

过表达CCR9对SW480结肠癌细胞侵袭的影响 　　[摘要] 目的 初步探讨人CCR9基因对SW480结肠癌细胞侵袭功能的影响。 方法 取人外周血单个核细胞提取总RNA逆转录为cDNA，扩增提取CCR9的CDS序列，连接pLVX-puro质粒，Lipo2000感染人SW480结肠癌细胞，嘌呤霉素筛选并建立稳定转染细胞株。免疫印迹证实CCR9基因在SW480结肠癌细胞高效表达。 Transwell方法检测SW480结肠癌细胞侵袭。 结果 免疫印迹证实成功构建pLVX-puro-CCR9质粒，相比SW480细胞组，CCR9过表达SW480结肠癌细胞中CCR9显著表达[（26.0±10.7） vs （10.0±4.5），n=8，P=0.0078]，且在体外能够促进SW480结肠癌细胞侵袭（P=0.0136）。 结论 成功构建CCR9稳定表达结肠癌细胞株，并能够在体外显著促进其侵袭功能。 　　[关键词] 趋化因子受体；CCR9；分子克隆；侵袭；SW480结肠癌细胞 　　[中图分类号] R735.35 [文献标识码] A [文章编号] 1673-9701（2016）25-0001-03 　　Influence of over-expressed CCR9 on invasion of SW480 colon cancer cells 　　LIU Haiyan1 XI Weixing1 HONG Ying1 CHAI Rui2 　　1.Department of Clinical Laboratory， Zhejiang Provincial Tongde Hospital， Hangzhou 310012， China； 2.Department of Anorectal Surgery， Zhejiang Provincial People’s Hospital， Hangzhou 310014， China 　　[Abstract] Objective To primarily explore the influence of human CCR9 gene on invasion function of SW480 colon cancer cells. Methods PBMC total RNA was extracted and reversely transcribed as cDNA， then amplified to extract CDS sequence of CCR9， which was then connected to pLVX-puro plasmid and Lipo2000 to infect human SW480 colon cancer cells， and screened by puromycin to establish stable transfection cell line. The over-expression of CCR9 gene in SW480 colon cancer cells was confirmed by western blotting. Transwell was used to detect invasion of colon cancer cells. Results PLVX-puro-CCR9 plasmids were successfully established as confirmed by western blotting. Compared with the SW480 cell group， CCR9 was significantly expressed in the CCR9 over-expression SW480 colon cancer cells [（26.0±10.7） vs （10.0±4.5）， n=8， P=0.0078]， and could facilitate invasion of SW480 colon cancer cells in vitro（P=0.0136）. Conclusion Colon cancer cell line with stable-expressed CCR9 can be successfully established， which can significantly facilitate the invasion function in vitro. 　　[Key words] Chemokine receptor； CCR9； Molecular clone； Invasion； SW480 colon cancer cell 　　趋化因