趋化因子受体CXCR5促进人乳腺癌MCF-7细胞的侵袭和转移

Investigate invasion and migration effect of CXCR5 on breast MCF-7 cancer cell

目的 构建在人乳腺癌MCF-7细胞中高效表达趋化因子受体CXCR5的质粒并检测CXCR5在MCF-7细胞中蛋白表达,探索CXCR5体外是否促进人乳腺癌MCF-7细胞的侵袭和转移。方法 提取外周血分离单个核细胞,抽提总RNA并逆转录为c DNA,构建CXCR5的真核质粒CXCR5-p LVX-IRES-puro。通过脂质体转染人乳腺癌MCF-7细胞。免疫印迹检测CXCR5质粒的人乳腺癌MCF-7细胞中CXCR5蛋白。Transwell检测MCF-7细胞和表达CXCR5的MCF-7细胞的侵袭和转移。结果 CXCR5基因成功构建至p LVX-IRES-puro质粒中,免疫印迹提示CXCR5在人乳腺癌MCF-7细胞中过度表达。Transwell结果显示CXCR5显著促进人乳腺癌MCF-7细胞的侵袭和转移。结论 在人乳腺癌MCF-7细胞中成功表达CXCR5基因,且促进人乳腺癌MCF-7细胞的侵袭和转移。

Objective To construct human CXCR5 gene over-express plasmid which encodes CXCR5 gene in human MCF-7 breast cancer cells,and to investigate the invasion and migration function of CXCR5 gene. Methods Peripheral blood was extracted and mononuclear cells were isolated,total RNA was extracted and reverse transcribed into c DNA,and the eukaryotic plasmid( CXCR5-p LVX-IRES-puro) of CXCR5 was constructed. Human breast cancer MCF-7 cells were transfected by liposome. CXCR5 protein of human breast cancer MCF-7 cells in CXCR5 plasmid was detected by Western blotting. Transwell was used to detect the invasion and metastasis of MCF-7 cells and MCF-7 cells expressing CXCR5. Results The CXCR5 gene was successfully constructed into the p LVX-IRES-puro plasmid,and immunoblotting indicated that CXCR5 was overexpressed in human breast cancer MCF-7 cells. Transwell results showed that CXCR5 significantly promoted invasion and metastasis of human breast cancer MCF-7 cells. Conclusion The CXCR5 gene was successfully expressed in human breast cancer MCF-7 cells and can promote their invasion and migration.