摘要
目的检测不同剂量顺铂(DDP)对人宫颈癌SiHa细胞增殖、凋亡及周期的影响,探讨DDP治疗宫颈癌的作用机制。方法采用WST-8方法检测DDP对人宫颈癌SiHa细胞增殖的影响;Hoechst33258荧光染色法检测细胞凋亡形态;用流式细胞仪检测药物作用前后细胞凋亡和周期变化。结果 DDP对SiHa细胞增殖的抑制作用,一定程度上呈浓度依赖性和时间依赖性,但0.01～1.00μg/ml的DDP作用24h和0.01～0.10μg/ml的DDP作用48h对细胞的抑制增殖作用不明显(P>0.05)。5.00μg/ml的DDP作用24h时,Hoechst33258荧光染色可见典型的凋亡细胞形态;1μg/ml的DDP作用SiHa细胞24h时,使SiHa细胞产生明显的S期阻滞,S期细胞比率为(53.17±7.50)%,与对照组、DDP5.00μg/ml组比较差异均具有统计学意义(P<0.05),致凋亡作用不明显(P>0.05);而5.00μg/mlDDP作用24h时,细胞早期凋亡率、晚期凋亡和坏死率分别为(5.88±1.64)%和(5.18±1.26)%,与对照组、DDP1μg/ml组比较差异均具有统计学意义(P<0.05)。结论 DDP抑制宫颈癌SiHa细胞增殖,但对小剂量DDP存在一定的耐药,其耐药机制可能与S期阻滞有关;高剂量DDP抗宫颈癌细胞作用机制主要表现为致凋亡作用。
Objective To investigate the possible mechanism of anticancer action of Cisplatin （DDP） in vitro by studying the effect of DDP in different diluted degrees on the cell proliferation,apoptosis and cell cycle and of SiHa cell line from human cervical carcinoma.Methods The effect of DDP on the SiHa cell line from human cervical carcinoma in vitro was detected by WST-8 assay,the apoptosis morphologic feature was assessed by Hoechst 33258 staining,and the apoptosis and cell cycle arrest were analyzed by flow cytometry.Results DDP could significantly inhibit the proliferation of SiHa cell in partly a concentration-dependent and time-dependent manner.But the inhibitory effect was not evident when it was 0.01-1.0μg/ml DDP for 24h and 0.01-0.1μg/ml DDP for 48h （P0.05）.Hoechst 33258 staining showed that typical appearance of cell apoptosis could be found in 5μg/ml DDP for 24h.The S cell cycle arrest of SiHa cell line could be found in 1μg/ml DDP for 24h,and the S cell proportion was （53.17±7.50）%,which was significantly different from that in the 5μg/ml DDP and control groups （P0.01）,with no obviously apoptosis-induced effect by 1μg/ml DDP （P0.05）.And the rates of early apoptosis,late apoptosis and sclerosis were （5.88±1.64）%,and （5.18±1.26）% respectively in 5μg/ml DDP for 24h,which were significantly higher than those in the 1μg/ml DDP and control groups（P0.05）.Conclusion DDP could inhibit the proliferation in SiHa cells,but SiHa cells showed resistant to lower doses of DDP,which may be caused by S cell cycle arrest.The possible mechanism of anticancer action of higher doses of DDP in vitro was mainly inducing apoptosis.
出处
《中国全科医学》
CAS
CSCD
北大核心
2010年第15期1615-1617,共3页
Chinese General Practice
基金
温州市科技局计划项目(Y20090157)
国家自然科学基金项目资助(30700195)
关键词
宫颈肿瘤
顺铂
细胞增殖
细胞周期
细胞凋亡
化疗敏感性
Uterine cervical carcinoma
Cisplatin
Cell proliferation
Cell cycle
Cell apoptosis
Chemosensitization
