卵巢癌细胞系CAOV3体外化疗后survivin mRNA表达的变化

Alteration of survivin gene expression in ovarian cancer cell line CAOV3 after chemotherapy in vitro

目的 探讨卵巢癌细胞系CAOV3体外化疗后survivinmRNA表达的变化。方法 采用四甲基偶氮唑蓝 (MTT)比色法检测不同浓度 (分别为 2 0 0 0、2 0 0、2 0、2、0 2mg/L)紫杉醇或卡铂对CAOV3细胞体外生长的抑制作用。应用RT PCR技术检测高浓度紫杉醇或卡铂 (分别为 2 0 0、5 0 0mg/L)处理 1d ,低浓度紫杉醇或卡铂 (分别为 2 0、5 0mg/L)处理 1、3、5d后存活的卵巢癌细胞中survivinmRNA的表达水平 ,同时利用流式细胞仪检测细胞凋亡情况。结果 不同浓度 (分别为 2 0 0 0、2 0 0、2 0、2、0 2mg/L)紫杉醇或卡铂作用后CAOV3细胞的抑制率分别为 94 %、88%、34%、2 2 %、13%或 97%、4 6 %、14 %、9%、7% ,随着药物浓度的下降 ,细胞受抑制程度明显减弱 (P <0 0 5 )。低浓度紫杉醇处理1、3、5d后CAOV3细胞的凋亡率分别为 15 %、2 3%和 2 9% ;高浓度紫杉醇处理 1d后CAOV3细胞的凋亡率为 4 3%。低浓度卡铂处理 1、3、5d后CAOV3细胞的凋亡率分别为 14 %、2 2 %和 2 6 % ;高浓度卡铂处理 1d后CAOV3细胞的凋亡率为 19%。紫杉醇或卡铂低浓度处理 3d及高浓度处理 1d后存活的卵巢癌细胞中survivinmRNA的表达量均明显高于未加药的对照 (P <0 0 1) 。

Objective To study the alteration of survivin gene expression in ovarian cancer cell CAOV3 after chemotherapy in vitro. Methods After the secondary culture of ovarian cancer cell line CAOV3, inhibition of ovarian cancer cell growth induced by paclitaxel or carboplatin at different concentration was detected by methyl thiazolyl tetrazolium(MTT). Alteration of survivin mRNA expression was evaluated by reverse transcription polymerase chain reaction (RT-PCR) in ovarian cancer cells which had been cultured with paclitaxel or carboplatin of high concentration (200, 500 mg/L) for 1 d or of low concentration (20, 50 mg/L) for 1, 3 and 5 d, respectively. Meanwhile, CAOV3 cell apoptosis was detected by flow cytometry. Results The inhibition rates on growth of ovarian cancer cell CAOV3 by different concentrations of paclitaxel or carboplatin after 48 h were 94%, 88%, 34%, 22%, 13% and 97%, 46%, 14%, 9%, 7%. The apoptosis rates of CAOV3 in low concentration paclitaxel for 1, 3, and 5 d were 15%, 23% and 29%. And the apoptosis rate in high concentration paclitaxel for 1 d was 43%. The apoptosis rate of CAOV3 in low concentration carboplatin for 1, 3, and 5 d were 14%, 22% and 26%. And in high concentration carboplatin for 1 d the rate was 19%. Expression of survivin mRNA increased distinctly after culturing in paclitaxel or carboplatin of low concentration for 3 d and of high concentration for 1 d compared to the control group ( P <0.01), the increase was especially evident in paclitaxel treated group ( P <0.01). Conclusion Survivin gene may play an inhibitory role in the chemotherapy of ovarian cancer cell CAOV3 in vitro.