罗格列酮诱导卵巢癌SKOV3细胞凋亡过程中hTERT表达变化的研究

Study on Changes in Expression of Human Telomerase Reverse Transcriptase during Rosiglitazone Inducing Apoptosis of Human Ovarian Cancer Cell Line SKOV3

目的探讨罗格列酮对卵巢癌细胞株SKOV3细胞的凋亡作用,及细胞凋亡与端粒酶反转录酶(hTERT)之间的关系。方法使用倒置相差显微镜观察药物作用前后细胞形态变化;不同浓度罗格列酮作用24、48 h后,流式细胞仪分别检测卵巢癌SKOV3细胞凋亡情况;免疫荧光检测hTERT蛋白表达的变化;反转录-聚合酶链反应(RT-PCR)检测hTERT mRNA变化。结果罗格列酮处理后,倒置相差显微镜检测到卵巢癌细胞变圆,胞质固缩,细胞间隙增宽,部分细胞脱壁并漂浮。25～200μmol/L罗格列酮作用于细胞24 h后即发生细胞凋亡,48 h后凋亡更加明显,同时存在时间和浓度依赖性。药物作用48 h后免疫荧光检测hTERT蛋白表达下调,RT-PCR检测hTERT mRNA的表达下调,呈浓度依赖性。结论罗格列酮可以诱导卵巢癌SKOV3细胞株凋亡,具有时间浓度依赖性,其机制可能与下调hTERT的表达有关。

Objective To investigate the apoptosis effect of rosiglitasone on human ovarian cancer cell line SKOV3 and to elucidate the relationship between apoptosis and the expression of human telomerase reverse transcriptase（hTERT）. Methods Inverted phase contrast microscopy（IPCM）was used to observe morphologic changes in the cells. After different concentrations of rosiglitazone were administered 24h and 48h, flow cytometry （FCM）was conducted to measure apoptosis. Fluorescence microscopy was used to detect the expression levels of hTERT protein, and the expression levels of hTERT mRNA were detected by reverse transcription polymerase chain reaction （ RT-PCR）. Results After treatment of rosiglitazone,inverted phase contrast microscope detected ovarian cancer cells became round, cytoplasm shrunk, intercellular space grew wider,some cells broke away from the cultivating flask and floated. Under the action of the 25-200μmol/L rosiglitazone, ovarian cancer cell line SKOV3 occurred apoptosis after 24h, and was more obvious after 48h, in a time-and-dose-dependent manner. The expression of hTERT protein by rosiglitazone treatment after 48h was down-regulated detected by fluorescence microscopy, and the hTERT mRNA expression was also down-regulated ,both of which were dose-dependent decreases. Conclusion Rosiglitazone can significantly induce apoptosis of human ovarian cancer cell line SKOV3 which indicates a time-and-dose-dependent manner, and the mechanism may be related to a down-regulation of hTERT expression.