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[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.
[Objective] To investigate the inhibitory effect of the active ingredients of Daphnip rats against human cervical cancer Hela cell line cultured in vitro. [Methods] MTT assay was used to detect the inhibitory effect of the active ingredients from Daphne vaporariorum on the proliferation of Hela cell lines cultured in vitro and its cells The morphological changes of Hela cells were observed by inverted microscope, AO staining and ethidium bromide (EB) staining respectively. The cell cycle distribution of Hela cells was detected by flow cytometry. Cytochemistry was used to detect the expression of proliferating cell nuclear antigen Ki-67, cyclinD1 and candidate tumor suppressor gene Syk. [Results] MTT colorimetric assay showed that the active ingredients of Daphnip was different from that of Hela cell Inhibition of proliferation, and showed a significant concentration-dependent and time-dependent.Inverted microscopy, AO / EB staining observed, Bat Apoptotic activity of Hela cells showed a series of typical characteristics of apoptotic cells.Flow cytometry showed that (P <0.01), while the proportion of cells in S phase and G2 / M phase was significantly decreased (P <0.01), while the percentage of cells in G0 / The results of immunocytochemistry showed that Syk expression was increased and Ki-67 and CyclinD1 protein expressions were decreased in the dosing group compared with the control group (P <0.01). The difference was statistically significant (P < P <0.01). [Conclusion] The active ingredients from Daphne vaporariorum obviously inhibit the growth of human cervical cancer Hela cells and lead to the arrest of G0 / G1 phase. The cell cycle arrest of G0 / G1 cells may be the active component of bat One of the mechanisms of tumor function.