紫花牡荆素体外抑制人宫颈癌HeLa细胞增殖的研究

Inhibition of proliferation of human cervical cancer HeLa cells by casticin in vitro

背景与目的:紫花牡荆素(Casticin)是一种从蔓荆子中提取的具有广泛药理活性的多甲基黄酮化合物。有研究表明Casticin能诱导多种肿瘤细胞凋亡,但其对人宫颈癌细胞的作用尚缺乏文献资料。本研究旨在探讨Casticin抑制体外培养人宫颈癌HeLa细胞增殖作用及其机制。方法:MTT法检测Casticin对人宫颈癌HeLa细胞活性的抑制作用;平皿集落形成法检测Casticin对HeLa细胞集落形成率的影响;流式细胞术(FCM)检测Casticin对HeLa细胞周期的影响;采用Western blot分析蛋白表达的变化。结果:Casticin对人宫颈癌HeLa细胞增殖活性有较强的抑制作用,呈剂量和时间依赖性,作用48h的IC50值为2.82μg/mL;与对照组比较,细胞集落形成率明显下降(P<0.05);Casticin作用48h,以浓度依赖方式阻滞HeLa细胞于G2/M期,同时降低CyclinB1蛋白表达,增高P21蛋白表达。结论:Casticin抑制HeLa细胞的增殖作用可能与其降低CyclinB1蛋白表达、活化P21蛋白有关。

Background and purpose： Casticin, a polymethoxy flavanoid, isolated from Vitex trifolia L, has been reported to inhibit multiple cancer cells proliferation, but there was no report concerning cervical cancer. This study was aimed to investigate the effect of Casticin on inhibition of the proliferation of human cervical cancer HeLa cells in vitro and its mechanisms. Methods： Human cervical HeLa cells were cultured in vitro. The inhibitory effect of Casticin on the viability of human cervical cancer HeLa cells was evaluated through MTT assay. The colony formation was detected by plate colony formation assay and the distribution of cell cycles were analyzed by flow cytometry. The expressions of proteins related to cell cycle arrest were analyzed by Western blot. Results： Casticin significantly inhibited the viability of human cervical cancer HeLa cells in a dose-dependent and time-dependent manner （The IC50 was 2.82 μg/mL for 48 h）. The colony-forming rate was reduced drastically compared with the control group （P〈0.05）. The cells were noticeably accumulated in the G2/M phase in a dose-dependent manner by flow cytometric analysis after treatment with Casticin for 48 h. Western blot showed that expression of the P21 protein was up-regulated and protein levels of Cyclin B 1 was depressed in a concentration-dependent manner after treatment with Casticin for 48 h. Conclusion： Down-regulation of Cyclin B 1 protein expression and activation of P21 proteins were the possible mechanisms where Casticin could inhibit the proliferation of human cervical cancer HeLa cells in vitro.