黑色素瘤生长趋化因子1对卵巢癌细胞生物学功能的影响

Effect of CXCL1 gene expression on the biological function of epithelial ovarian carcinoma cells in vitro

目的探讨黑色素瘤生长趋化因子1（CXCL1）基因在卵巢癌细胞生物学功能中的作用。方法采用逆转录聚合酶链反应（RT．PCR）扩增CXCL1基因全长序列，并构建其表达的重组慢病毒表达载体。采用RT—PCR和酶联免疫吸附试验（ELISA）法检测慢病毒介导的卵巢癌A2780细胞中CXCL1mRNA和蛋白表达情况；采用四甲基偶氮唑蓝（MrITI．）法、流式细胞术和显微镜下计数集落数法测定细胞生长曲线、细胞生长周期和细胞集落形成情况；采用Transwe11小室和Migration侵袭方法测定细胞体外的迁移和侵袭能力。结果测序结果显示，重组慢病毒质粒CXCL1-PWPI与CXCL1基因的同源性达100％。lit-PCR和ELISA法检测结果显示，A2780细胞经CXCL1-PWPI转染后，CXCL1mRNA和蛋白表达呈阳性。A2780组、PWPI组和CXCL1-PWPI组的细胞集落形成率分别为22．3％、26．5％和37．2％（P〈0．05），G1期细胞分别为62．7％、61．4％和44．0％（P〈0．001）。CXCL1-PWPI组与A2780组和PWPI组比较，其细胞迁移能力差异无统计学意义（P〉0．05），但侵袭能力差异有统计学意义（P〈0．05）。结论CXCL1基因具有促进卵巢癌A2780细胞增殖和侵袭的能力。

Objective To investigate the effect of CXCL1 gene expression on biological function of ovarian cancer cells and its mechanism of action. Methods RT-PCR was used to amplify the full-length sequence of CXCL1, which was used to construct the recombinant lentiviral plasmids, and then the plasmids were packaged with human renal epithelial cell line 293T cells, and the ovarian cancer 2780 cells were transfected with CXCL1 gene by virus supernatant particles. The expression of CXCL1 mRNA and protein in 2780 cells with lentivirus-mediated CXCLlexpression were determined by RT-PCR and ELISA, respectively. The growth curve, cell cycle and colony formation in vitro were measured by MTT assay, flow cytometry and colony formation counting, respectively. The cell invasion and migration in vitro was detected by Transwell chambers and migration attack methods, respectively. Results The sequencing results showed that the recombinant lentiviral plasmid of CXCLI-pWPI had 100% homology with the CXCL1 gene, identified by BLAST analysis. RT-PCR and ELISA results showed positive expression of CXCL1 mRNA and protein in the CXCL1-PWPI group. The doubling time of the CXCL1-PWPI group was significantly shorter than that in the A2780 and PWPI groups （ P 〈 0.05 ）. The rate of colony-formation in the CXCL1-PWPI group was also significantly higher than that of the A2780 and PWPI groups （ P 〈 0.05 ）. The proportion of cells in G1 phase （44.0%） in the CXCL1-PWPI group was also significantly lower compared with that in the PWPI and A2780 groups （61.4% and 62.7% ）, with a significant difference （P 〈 0. 001 ）. There was no significant difference between the cell migration capacity （ P 〉 0.05 ） of the CXCL1-PWPI, PWPI and A2780 groups, but the invasion capacity of the CXCL1-PWPI group was significantly higher than that of the PWPI and A2780 groups （ P 〈 0.05 ）. Conclusion The over-expression of CXCL1 gene may promote the proliferation and invasion ability of A2780 cells in vitro.