不同浓度的盐酸千金藤碱对人鼻咽癌细胞的抑制作用实验研究

An experimental study of effects of cepharanthine hydrochloride on nasopharyngeal carcinoma cells

目的研究探讨不同浓度盐酸千金藤碱对人鼻咽癌细胞CNE-1/CNE-2和5-8F的抑制作用。方法选用鼻咽癌高分化鳞癌细胞株CNE-1和鼻咽癌低分化鳞癌细胞株CNE-2、5-8F作为实验组,人脐静脉内皮细胞株EVC-304为正常细胞对照组,利用MTT法检测不同浓度的盐酸千金藤碱对3种鼻咽癌细胞和对照组细胞的增殖影响;利用Hoechst 33342染色法检测不同浓度的盐酸千金藤碱对3种鼻咽癌细胞和对照组细胞的凋亡影响;利用流式细胞术检测盐酸千金藤碱对3种鼻咽癌细胞和对照组细胞的细胞周期影响。结果不同浓度的盐酸千金藤碱对3种鼻咽癌细胞有量效相关的抑制增殖作用,EC50分别为9.924μg/ml(CNE-1),7.068μg/ml(CNE-2),9.905μg/ml(5-8F),浓度为50μg/ml时抑制率达到95%以上,与对照组正常细胞相比差异均有统计学意义(P<0.01);不同浓度的盐酸千金藤碱3种鼻咽癌细胞有量效相关的诱导凋亡的作用,到12.5μg/ml时,处理组肿瘤细胞与阴性组相比染色质浓缩成斑块状,差异显著(P<0.05);同时,12.5μg/ml终浓度的盐酸千金藤碱处理不同的鼻咽癌细胞48 h后,与阴性对照组相比,可见实验组G0/G1期鼻咽癌细胞显著增多,S期及G2/M期明显减少,差异有统计学意义(P<0.05),表明盐酸千金藤碱能抑制其DNA合成,改变细胞周期。结论盐酸千金藤碱对3种鼻咽癌细胞均有抑制作用,可能机制包括诱导其凋亡并且改变其细胞周期。

Objective To study the inhibitory effect of different concentration of cepharanthine hydrochioride on nasopha- ryngeal carcinoma cell lines CNE-1, CNE-2 and 5-8F. Methods The proliferation, apoptosis and cell cycle of nasopha- ryngeal carcinoma cells （CNE-1, CNE-2,5-8F, experimental group） and normal cell line （EVC-304, control group） treated with cepharanthine hydrochloride were detected by M33＂, Hoechst 33342 and flow cytometry,respectively. Results Different concentrations of cepharanthine hydrochloride showed a dose-related inhibition of proliferation and induction of apoptosis on three nasopharyngeal carcinoma cell lines, EC50 of cepharanthine hydrochiorid on three cell lines were 9. 924 μg/ ml（ CNE-1 ） ,7. 068 μg/ml（CNE-2） and 9. 905 μg/ml（5-8F）, respectively. Inhibition rate of concentration at 50 μg/ml reached 95% which showed significant differences as compared the control group with normal cells（P 〈 0. 01 ）. Different concentrations of cepharanthine hydrochioride showed a dose-related inhibition of induction of apoptosis on 3 nasopharyngeal carcinoma cell lines too, after the treatment with 12.5 μg/ml cepharanthine hydrochioride, the chromatin condensed into a patch while which not observed in the control groups （ P 〈 0. 05 ）. Meanwhile cepharanthine hydrochiorid also changed their cell cycle by inhibition of DNA synthesis at 12.5 μg/ml （ P 〈 0.05 ）. Conclusion Cepharanthine hydrochloride can inhibited nasopharyngeal carcinoma cell lines by inducing apoptosis and changing cell cycle.