顺铂对卵巢癌SKOV3细胞水通道蛋白5表达的影响及调控

Cisplatin on the Effect of Aquaporin 5 Protein Expression of SKOV3 and Its Regulation

应用噻唑蓝法、蛋白质印迹和RT-PCR法研究顺铂对卵巢癌SKOV3细胞水通道蛋白5(AQP5)表达的影响及其调控。结果显示顺铂浓度增加,SKOV3细胞AQP5、胞浆和胞核NF-κBp65以及胞核IκBα表达均逐渐减少(P=0.001,0.000,0.000,0.000);10μg/ml顺铂与SKOV3细胞温育6～12h,AQP5、胞浆和胞核NF-κBp65以及胞浆IκBα表达均明显增加达峰值,24h后急剧下降到低值,并维持至72h后(P=0.000,0.000,0.000,0.000,0.000)。随着核转录因子阻断剂PDTC浓度增加、作用时间延长,AQP5表达及mRNA量均逐渐减少(P=0.000,0.000);顺铂对细胞抑制率与AQP5呈负相关(r=-0.598;P=0.009),PDTC对细胞抑制率与AQP5和mRNA呈负相关(r=-0.983,-0.905;P=0.000,0.000)。AQP5表达与NF-κBp65和IκBα呈正相关(r=0.894,0.857;P=0.000,0.000)。提示AQP5与SKOV3细胞生长有关,顺铂下调AQP5表达,其过程可能受NF-κB调控,为AQP5成为卵巢癌治疗的靶目标提供了一定的理论基础。

To investigate cisplatin on effect of aquaporin 5 （AQP5） expression of SKOV3 and regulation of PDTC that is NF-r,B inhibitor by MTT, Western blot and RT-PCR. Following the concentration of cisplatin increasing, the expression of AQP5 protein （P=0.001） as well as NF-κB p65 in cytoplasm and karyon and IκBα in karyon, P=0.000, 0.000, 0.000, in SKOV3 incubated with cisplatin 24 h decrease. After SKOV3 incubated with 10 μg/ml cisplatin 6 h to 12 h, the protein expression of AQP5, NF-κBB p65 in cytoplasm and karyon and It, Bet in cytoplasm increase rapidly, but decrease suddenly at 24 h, remain the lower level until 72 h （P=0.000, 0.000, 0.000, 0.000）. The more concentration and longer incubated time of PDTC, the more decreasing of AQP5 and mRNA （P=0.000, 0.000）. There is a negative correlation between AQP5 protein and the inhibiting rate of SKOV3 induced by cisplatin （r=-0.598, 0.009）, and a negative correlation between the inhibiting rate induced by PDTC and AQP5 （r=-0.983, P=0.000） as well as AQP5 mRNA （r=-0.905, P=0.000）, and a positive correlation between AQP5 expression and NF-κBB p65 as well as IκBα （r=0.894, 0.857; P=0.000, 0.000）. These results suggest that AQP5 is related to SKOV3 cells proliferation and cisplatin may reduce AQP5 expression, which may involved in NF-κB gene regulation, AQP5 will be potential target for chemotherapy of ovarian carcinoma.