蝙蝠葛活性成分对Hela细胞株的抑制增殖作用

目的]探讨蝙蝠葛活性成分对体外培养的人宫颈癌Hela细胞株的抑制增殖作用.方法]应用MTT法检测蝙蝠葛活性成分对体外培养的人宫颈癌Hela细胞株增殖抑制作用及其细胞毒活性;分别采用倒置显微镜、吖啶橙(AO)/溴化乙锭(EB)染色观察Hela细胞处理前后的形态学变化;采用流式细胞仪检测Hela细胞株的细胞周期分布相;应用免疫细胞化学法检测增殖细胞核抗原Ki-67、细胞周期蛋白CyclinD1及候选抑癌基因Syk的表达情况.结果]MTT比色法观察结果表明,蝙蝠葛活性成分对人宫颈癌Hela细胞株有明显的抑制增殖作用,且呈现明显的质量浓度和时间依赖性.倒置显微镜、AO/EB染色观察见,蝙蝠葛活性成分处理的Hela细胞出现一系列典型的凋亡细胞特征.流式细胞仪检测结果显示,蝙蝠葛活性成分作用后加药组出现G0/G1期细胞比例显著增加(P<0.01),而S期和G2/M期细胞比例则明显下降(P<0.01),将细胞阻滞在G0/G1期;免疫细胞化学观察结果表明,蝙蝠葛活性成分作用后加药组Syk表达增加,Ki-67和CyclinD1蛋白表达降低,与对照组相比较差异均有统计学意义(P<0.01).结论]蝙蝠葛活性成分对人宫颈癌Hela细胞株有明显的生长抑制作用,并可导致G0/G1期阻滞.G0/G1细胞周期阻滞可能是蝙蝠葛活性成分抗肿瘤作用的机制之一.

Antiproliferative effect of active components of menispermum dauricum on Hela cell line

OBJECTIVE To explore the proliferation inhibitory effect of active components of menispermum dauricum on Hela cell line.METHODS The cells of human cervical carcinoma cell line Hela were treated respectively at different concentration and time.The proliferation inhibitory rate and cytotoxic effect of Hela cells was measured by MTT assay;morphological changes of the apoptosis in cancer cells were investigated respectively with inverted microscope,acridine orange-ethidium bromide fluorescent staining(AO/EB) double fluorescent staining.The distribution of cells cycle and the apoptosis rate were observed by flow cytometry;the expression of nuclear proliferating antigen Ki-67,cell cycle proteins Cyclin D1 and candidate anti-oncogene Syk were detected by SP immunohistochemitry.RESULTS MTT assay showed that active components of menispermum dauricum could significantly inhibit the growth of Hela cells and the effect was in a dose-dependent and time-dependent manner(P<0.01);flow cytometry showed active components of menispermum dauricum could induce the Hela cells apoptosis;some typical sub-dipioid peaks were observed,the percentage of cells in G0/G1 phase increased and the percentage of cells in S and G2/M phase decreased after the cells treated by active components of menispermum dauricum;the active components of menispermum dauricum could arrest Hela cells in G0/G1phase;immunocytochemistry showed that the expression of candidate anti-oncogene Syk increased,the expression of nuclear proliferating antigen Ki-67 and cell cycle proteins Cyclin D1 decreased after the Hela cells treated by the active components of menispermum dauricum,there were different significantly compared with control(P<0.01).CONCLUSION The active components of menispermum dauricum could inhibit the proliferation of Hela cells in vitro.Its mechanism may be related with arresting cell cycle in G0/G1 phase.