CXCR2过表达结肠癌细胞株的建立及生物学功能的初步研究

目的 建立人SW1116结肠癌细胞中稳定过表达CXCR2稳定细胞株,分析过表达CXCR2基因对人结肠癌SW1116细胞迁移能力的影响。方法 分离健康人外周血单个核细胞,Trizol提取总RNA并反转录为cDNA,PCR扩增CXCR2的CDS序列,连接至慢病毒穿梭质粒pLVX-IRES-ZsGreen1多克隆位点,进行CXCR2基因的克隆,构建pLVX-IRES-ZsGreen1-CX-CR2重组质粒。重组质粒与包装质粒通过磷酸钙共转染法包装出慢病毒上清并对人SW1116结肠癌细胞进行感染。real-timePCR及Western blot方法分析转染pLVX-IRES-ZsGreen1-CXCR2重组质粒的人SW1116结肠癌细胞中CXCR2表达情况。Tr-answell实验分析过表达CXCR2对结肠癌细胞体外迁移的影响。结果 成功建立稳定表达CXCR2基因的SW1116结肠癌细胞株,并初步阐明CXCR2能够促进结肠癌细胞体外迁移。结论 成功建立过表达CXCR2基因的结肠癌细胞并能促进其体外迁移。

Establish Ment of CXCR2 Overexpression Human Colorectal Cancer Cell Line and Its Effect on the Bilogical Function

Objective To establish the stable overexpressive human CXCR2 gene in human colorectal cancer SW1116 cells.Methods The CXCR2 cDNA was amplified with primers from the total RNA of the PBMC of the health person and then cloned to CDS sites of the pLVX-IRES-ZsGreen1 lentivirus shuttle vector.The lentivirus shuttle plasmid and package plasmid were con tramsfected into HEK293T cell to produce the CXCR2 lentivirus,then the CXCR2 overexpress SW1116 cell line was established.The realtime-PCR and westernblot wore used to detect the CXCR2 expression in the stable CXCR2 overexpress SW1116 cell line.The transwell assay were used to evaluate the effect of overexpression of CXCR2 on the migration ability of SW1116 colorectal cancer cell line.Results We have successfully established CXCR2 overexpress SW1116 cell line and demonstrate that CXCR2 can facilitate the colorectal cancer migration in vitro.Conclusion CXCR2 overexpress SW1116 cell line had established and it can facilitate the migration in vitro.