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作 者:李桂玲[1] 归绥琪[1] 王莉[1] 林理根[2] 叶阳[2]
机构地区:[1]复旦大学附属妇产科医院中西医结合科,上海200011 [2]中国科学院上海药物研究所天然药化研究室
出 处:《中国中西医结合杂志》2010年第3期303-307,共5页Chinese Journal of Integrated Traditional and Western Medicine
基 金:上海市科委科研计划项目(No.074119613)
摘 要:目的观察中药掌叶半夏有效提取物(Pinellia extract,PE)对HeLa细胞的作用,并对可能的机制进行初步研究,为其临床用于防治女性宫颈病变提供科学依据。方法用倒置相差显微镜观察细胞生长并摄影,MTT法检测细胞活性,流式细胞术检测细胞凋亡,免疫细胞化学法检测增殖细胞核抗原(proliferatingcell nuclear antigen,PCNA)蛋白表达,免疫荧光实验检测B细胞淋巴瘤/白血病-2(Bcl-2)蛋白表达。结果PE对HeLa细胞的生长表现出明显的抑制作用,流式细胞术检测结果显示PE作用后HeLa细胞出现凋亡,且随着作用时间延长,凋亡细胞的比例明显增加;免疫细胞化学实验和免疫荧光实验结果表明:PE作用24 h后HeLa细胞PCNA和Bcl-2蛋白表达均明显减少。结论PE对HeLa细胞有明显的抑制生长和促进凋亡的作用,其促凋亡机制是通过降低PCNA和Bcl-2蛋白表达来实现的,这为进一步研究PE的作用机制以及抗癌新药的研制开发奠定了初步的实验基础。Objective To investigate the effect of Pinellia extract (PE) on HeLa cell line and to study its associated mechanisms, in order to provide theoretical foundations for its applying in clinical prevention and treatment of cervical disease. Methods HeLa cell line was incubated in media containing different concentrations of PE. Growth of cells was observed and photo-generated with inverted phase microscope; cell activity was detected by MTT assay; cell apoptosis detected by flow cytometry, protein expression of proliferating cell nuclear antigen proliferating cell nuclear antigen (PCNA) was detected by immuno-cytochemistry, and protein expression of Bcl-2 was determined by immunofluorescence. Results PE showed obvious inhibition on the proliferation of HeLa cells, cell apoptosis appeared after PE treatment in a time and dose dependent manner; PCNA and Bcl-2 protein expressions reduced significantly after being effected by PE for 24 h. Conclusions PE can obviously inhibit the growth and induce the apoptosis of HeLa cells, and its mechanism is possibly realized through down-regulating the expressions of PCNA and Bcl-2. The study set a primary experimental base for further studying the action mechanism of PE and developing new anti-tumor agents.
关 键 词:掌叶半夏有效提取物 宫颈癌 HELA细胞株 增殖细胞核抗原 B细胞淋巴瘤/白血病-2
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