拓扑替康对卵巢癌SKOV3细胞水通道蛋白5及NF-κB表达的影响

Effect of topotecan on expression of aquaporin protein 5 and nuclear factor-κB in ovarian cancer SKOV3 cells

背景与目的:水通道蛋白5(aquaporin 5,AQP5)的表达异常与卵巢癌细胞的浸润转移和血管形成有关。核转录因子NF-κB参与细胞的增殖、分化、凋亡,且能影响肿瘤细胞对化疗的敏感性。本研究探讨拓扑替康(topotecan,TPT)对卵巢癌SKOV3细胞中AQP5、核转录因子(nuclear factor kappaB,NF-κB)及其抑制因子(inhibitor kappa B,IκBα)表达的影响。方法:用不同浓度TPT作用SKOV3细胞不同时间后,应用MTT法检测并计算卵巢癌细胞增殖抑制率,蛋白质印迹法检测AQP5、NF-κB及IκBα蛋白表达,并分析各蛋白表达水平的相关性。结果:0.4μg/mLTPT与SKOV3细胞温育24h,AQP5、胞浆和胞核NF-κBp65以及胞浆IκBα表达随着作用时间延长均明显下降,并维持至72h后(P<0.05)。不同浓度(0.2μg/mL、0.4μg/mL、0.6μg/mL、0.8μg/mL)TPT作用于SKOV3细胞24h,随着TPT浓度增加,SKOV3细胞AQP5表达量逐渐减少(P<0.05),0.8μg/mLTPTAQP5表达抑制率为57.9%;同时,细胞核NF-κBp65、IκBα表达量也逐渐减少。TPT对细胞的增殖抑制率与AQP5表达呈负相关(r=-0.965;P<0.05),AQP5表达与NF-κBp65和IκBα呈正相关(r=0.903,0.896;P<0.05)。结论:TPT抑制肿瘤细胞增殖时下调AQP5及核转录因子NF-κB表达。

Background and Objective. Overexpression of aquaporin protein 5 （AQP5） is associated with the metastasis, migration and angiogenesis of ovarian cancer, but its function in ceil proliferation has not been described. This study was to explore the effects of topotecan （TPT） on the expression of AQP5, nuclear factor-KB （NF-κB） and its receptor IκBα in ovarian cancer SKOV3 cells. Methods： When SKOV3 cells were treated with TPT of various concentrations for different time, cell proliferation was measured by MTT assay, the expression of AQP5, NF-κB and IκBα was detected by Western blot. Results： After SKOV3 cells were incubated with 0.4μg/mL TPT for 24 h, the expression of AQP5, NF-κB p65 in cytoplasm and nuclei, and IκBα in cytoplasm were decreased, and remained at low levels till 72 h （P〈 0.05）. When SKOV3 cells were treated with 0.2, 0.4, 0.6 and 0.8 μg/mL TPT for 24 h, the expression of AQP5 as well as NF-κB p65 and IκBα in nuclei were decreased （P〈0.005）. The protein level of AQP5 was decreased by 57.9% when cells were treated with 0.8μg/mL TPT. AQP5 expression was negatively correlated to the proliferation inhibition rate of SKOV3 cells induced by TPT （r=-0.965, P〈0.05）, and positively correlated to NF-κB p65 and IκBα expression （r=0.903, 0.896, P〈0.05）. Conclusion. When inhibiting the proliferation of ovarian cancer cells, TPT may down-regulate the expression of AQP5 and NF-κB.