TWEAK对人卵巢癌细胞HO-8910PM侵袭转移的影响及其机制的研究

目的:研究肿瘤坏死因子样凋亡微弱诱导剂(TWEAK)对人高转移卵巢癌细胞HO-8910PM转移相关能力的影响及其作用机制。方法:水溶性四唑盐(WST-1)比色法检测TWEAK对HO-8910PM细胞增殖力的影响,细胞黏附和移动实验检测TWEAK对细胞体外黏附和移动力的影响,细胞侵袭重建基底膜实验检测TWEAK组细胞体外侵袭力的变化。采用Western blot法检测TWEAK对细胞IκB-α、NF-κB(P65)和VEGF蛋白表达的影响。通过应用NF-κB拮抗剂PDTC,观察阻断NF-κB通路后VEGF蛋白表达的变化。结果:TWEAK组在24、48h和72h时细胞增殖率无明显变化(P>0.05)。TWEAK组细胞黏附力提高23.67%(P<0.01);TWEAK可增强细胞体外的移动和侵袭力,其趋化指数和侵袭指数分别为1.9±0.3和1.7±0.3(P<0.01)。TWEAK能显著下调IκB-α蛋白表达,上调NF-κB(P65)蛋白在核内的表达,并使VEGF蛋白表达量显著增加。用NF-κB拮抗剂PDTC阻断NF-κB通路,能有效抑制VEGF蛋白的表达。结论:TWEAK对HO-8910PM细胞增殖力无明显影响。TWEAK能促进HO-8910PM细胞移动、黏附和侵袭力。TWEAK促肿瘤侵袭转移的机制之一可能是通过激活NF-κB通路上调VEGF蛋白表达。

Effect and mechanism of TWEAK on the invasion ability of ovarian cancer cell line HO-8910PM

Objective:To investigate the effect and mechanism of TWEAK on the invasion ability of ovarian cancer cell line HO-8910PM.Methods:The proliferation rate of HO-8910PM cells was detected by WST-1 cell proliferation assay.The abilities of adhesion,migration and invasion were detected by cell adhesion assay,cell migration assay and Matrigel invasion assay respectively.The expressions of IκB-α、NF-κB(P65) and VEGF protein were observed by Western blot when HO-8910PM cells were treated with TWEAK at the different time.PDTC,the specific inhibitor of NF-κB pathway,was also used to inhibit NF-κB and to study the expression of VEGF protein.Results:The presence of TWEAK did not affect the proliferation rate of HO-8910PM cells.The adhesion of TWEAK-stimulated cells for 60min increased by 23.67%.TWEAK promoted the migration and invasion of HO-8910PM cells.The chemotactic index(CI) and invasion index(Ⅱ) were 1.9±0.3 and 1.7±0.3 respectively,significantly higher than those of the control.TWEAK induced the degradation of IκB-α protein.The levels of NF-κB(P65) protein in nucleus and VEGF protein were increased in the cells treated by TWEAK.Blocking the NF-κB pathway with PDTC neutralized the upregulation of VEGF protein expression induced by TWEAK.Conclusion:TWEAK can promote the adhesion,migration and invasion of the ovarian cancer cell HO-8910PM.One of the mechanisms of the effect is probably that TWEAK upregulates the expression of VEGF protein through activating NF-κB pathway.