NK4基因转染对人宫颈癌细胞CaSki生物学特性的影响

 目的 通过在人宫颈癌细胞CaSki中转染NK4 cDNA片段，研究NK4对人宫颈癌的潜在治疗作用。方法 将NK4和荧光霉素表达质粒分别稳定转染人宫颈癌细胞CaSki。采用Western blot检测细胞培养基中NK4蛋白以及经不同培养基（CaSki, CaSki/LUC 及CaSki/NK4培养上清）培养后CaSki细胞中c-Met和磷酸化c-Met的表达。采用MTT法绘制细胞生长曲线。采用细胞划痕试验检测NK4对细胞转移的作用。结果 在CaSki/NK4培养基中有NK4蛋白表达，对照细胞（CaSki, CaSki/LUC）中无表达。磷酸化c-Met的表达在CaSki/NK4培养基培养的CaSki细胞中被抑制，而在对照细胞（CaSki, CaSki/LUC）培养基培养的CaSki细胞中正常表达。各组c-Met表达无差异。3种细胞体外生长曲线差异无统计学意义（P >0.05）。细胞划痕实验结果表明，CaSki/NK4划痕区域的细胞个数明显少于CaSki和CaSki/LUC。结论 NK4能抑制人宫颈癌细胞c-Met受体磷酸化，并能抑制宫颈癌细胞体外活动能力。

Effects of NK4 on Biological Behaviors of CaSki

Objective To clarify the antitumor activity of NK4 in cervical cancer and investigate NK4 as a potential therapeutic method to cervical cancer.Methods NK4 cDNA was introduced into a human cervical cancer cell line CaSki.The effect of transfected NK4 gene on biological behaviors of CaSki were observed.Western blot analysis was used to detect NK4 expression in the supernatants of CaSki,CaSki/LUC and CaSki/NK4 cells and to detect c-Met and phospho-c-Met expression in CaSki cells cultured with supernatants from CaSki,CaSki/LUC and CaSki/NK4 cells.Migration capabilities of the cells were evaluated in vitro using scratch wound healing assay.Cell growth curves were performed using MTT kit.Migration capabilities of the cells were evaluated in vitro using scratch wound healing assay.Results NK4 was detected in the supernatant of CaSki/NK4 and NK4 expressed in the supernatant of CaSki/NK4 inhibited phosphorylation of c-Met in CaSki cells.There was no difference between CaSki/NK4 cells and control(CaSki or CaSki/LUC)cells in doubling time in vitro.The number of migrated cells was significantly smaller in the CaSki/NK4 cells than that of the control(CaSki or CaSki/LUC) cells.Conclusion NK4 could inhibit phosphorylation of c-Met in CaSki cells and inhibit cell migration capability of CaSki cells.But NK4 had no effect on cell proliferation in CaSki cells.These findings suggested NK4 had the potential to be a gene targeted therapeutic strategy to human cervical cancer.