顺铂抑制SKOV3细胞转移的动态监测及相关机制探讨

Real time analysis of the characteristic of SKOV3 cell metastasis inhibited by cisplatin and the correlative mechanism

目的:在细胞水平实时动态监测顺铂抑制SKOV3细胞黏附和转移特性,探讨其相关机制。方法:CCK8法检测顺铂对SKOV3细胞增殖的抑制;采用RTCA监测仪24h实时动态观察顺铂对SKOV3细胞的黏附、迁移和侵袭的抑制作用;细胞组化法检测顺铂作用SKOV3细胞24h后细胞表面CD44v6的表达情况。结果:(1)顺铂对SKOV3细胞增殖的抑制作用呈浓度和时间依赖性,24、48h的IC50分别为24.065±1.031、5.083±0.128μg/ml;(2)在第50min时点,25μg/ml顺铂组与对照组对SKOV3细胞黏附力的抑制作用差异最显著(P<0.05)。第2h时点,12.5、25μg/ml顺铂组对SKOV3细胞迁移的抑制作用显著高于对照组(P<0.05)。第10h 15min时点,25μg/ml顺铂组对SKOV3细胞侵袭的抑制作用显著高于对照组(P<0.05)。(3)与对照组比较,25μg/ml顺铂组细胞表面表达CD44v6明显减少(P<0.05),而5μg/ml顺铂组减少不明显。结论:顺铂抑制SK-OV3细胞转移特性首先表现为抑制细胞的黏附和迁移,这与顺铂减少细胞表面CD44v6蛋白的表达密切相关。CD44v6将成为上皮性卵巢癌治疗的新靶点。

Objective：To real time analyze the characteristic of SKOV3 cell adhesion and metastasis inhibited by cisplatin, and study the role of CD44v6 during the process. Meth- ods：The inhibitory effect of eisplatin on the proliferation of SKOV3 cells was evaluated by CCK8 kit. The features of SKOV3 cell adhesion, migration and metastasis were analyzed by real time cell analyzer for 24 hours. The expression of CD44v6 protein of SKOV3 cell after cisplatin effected for 24 hours was evaluated by IHC. Results： （ 1 ） Cisplatin significantly inhibited SK- OV3 cells in a dose-dependent and time-dependent manner, and the IC50 was （24. 065 ±1. 031 ）μg/ml for 24 hours, （ 5. 083±0. 128 ） μg/ml for 48 hours. （2） Cisplatin inhibited cell adhesion ： the cell index of 25 μg/ml cisplatin group at the first 50rain was lower than the control group （ P 〈0.05 ）. Cisplatin inhibited cell migration ：the cell index of 12.5,25μg/ml cisplatin groups at the first 2 hours was lower than the control group respectively （ P〈0.05 ）. Cisplatin inhibited cell invasion：the cell index of 25μg/ml cisplatin group at the first 10 hours 15rain was nealy 0, while its of the control group was 0. 7488_＋0. 1565 （ P〈0.05 ）. （3） Cisplatin decreased the ex- pression of CD44v6 of SKOV3 cells. The staining score of 25 μg/ml Cisplatin group was lower than the control group （ 1. 250 ±0. 250 vs 5. 250 ±0. 250, P〈0.05 ）. Conclusions： Cisplatin first- ly inhibit the adhesion and migration of SKOV3 cells during the tumor cell metastasis process, and it may be closely related with decreased the expression of CD44v6 of SKOV3 cells. It canbe novel target for ovarian cancer treatment.