摘要
目的探讨那可丁对宫颈癌HeLa细胞株的抑制作用及其机制。方法体外培养人宫颈癌HeLa细胞株,梯度浓度那可丁干预后,MTT测定细胞存活率,软琼脂克隆形成实验检测细胞成瘤能力,Western blot检测Caspase-3、PARP(多聚ADP-核糖聚合酶)蛋白水平变化,流式细胞术检测细胞凋亡率及细胞周期分布。结果 MTT检测发现那可丁显著降低HeLa细胞存活率(P<0.05),该效应具有浓度依赖性和时间依赖性;软琼脂克隆形成实验发现,10、20、30、40μmol/L那可丁组的细胞克隆计数依次为(19±3)、(13±2)、(4±1)(、3±1),均显著低于对照组的(62±8)(均P<0.01);流式细胞术检测显示那可丁显著增加细胞凋亡率(P<0.05),该效应具有浓度依赖性和时间依赖性;Western blot检测显示那可丁上调Caspase-3(17 kD)、PARP(85 kD)蛋白水平;流式细胞术检测显示那可丁阻滞HeLa细胞周期于G2/M期(P<0.05)。结论那可丁对HeLa细胞有显著抑制效应;那可丁诱导的G2/M期周期阻滞可能在该效应中发挥作用。
Objective To investigate the effects of noscapine on human cervical cancer cell line HeLa and its mechanism.Methods HeLa cells were cultured in vitro and treated with gradient concentrations of noscapine.MTT assay was used to detect the proliferation of HeLa cells.Soft-agar colony-forming assay was used to measure tumor-forming ability.Flow cytometry（FCM）was used to measure apoptosis and cell cycle distribution of HeLa cells.Western blot was used to detect protein levels of Caspase-3 and poly ADP-ribose polymerase（PARP）.Results MTT assay showed that noscapine decreased the survival rate of HeLa cells in a time-and dose-dependent manner（P〈0.05）.Soft-agar colony-forming assay displayed that clone number of HeLa cells treated with 10,20,30 or 40 μmol/L noscapine was（19±3）,（13±2）,（4±1） and（3±1） respectively,that was（62±8） in group control,which indicated a significantly decreased tumor-forming ability of HeLa cells（P〈0.01）.FCM analysis revealed that noscapine increased apoptosis rate of HeLa cells in a time-and dose-dependent manner（P〈0.05）.Western blot analysis demonstrated that noscapine increased protein levels of Caspase-3（17 kD）and PARP（85 kD）in HeLa cells.Noscapine arrested HeLa cells in G2/M phase（P〈0.05）.Conclusion Noscapine significantly inhibits HeLa cells.Noscapine-induced G2/M phase arrest may play an important role in the effects.
出处
《华中科技大学学报（医学版）》
CAS
CSCD
北大核心
2011年第3期287-290,共4页
Acta Medicinae Universitatis Scientiae et Technologiae Huazhong
基金
国家自然科学基金资助项目(No.30770942)
武汉市青年科技晨光计划资助项目(No.200750731294)
关键词
那可丁
宫颈癌
细胞周期
细胞凋亡
noscapine
uterine cervical neoplasms
cell cycle
apoptosis
