汉黄芩苷体外抗结肠癌作用及机制研究

In vitro anti-colorectal cancer effect of Wogonoside and its mechanism

目的研究汉黄芩苷对结肠癌细胞和肾小球系膜细胞增殖的影响,并探讨其诱导肿瘤细胞凋亡的机制。方法体外培养的结肠癌细胞和肾小球系膜细胞经不同浓度的汉黄芩苷处理后,MTT法检测细胞的存活率,激光共聚焦显微镜观察LoVo细胞的形态学变化,流式细胞仪术检测LoVo细胞的凋亡率,逆转录聚合酶链反应(RT-PCR)检测LoVo细胞凋亡相关基因bcl-2、bax的表达情况。结果 MTT检测结果显示,汉黄芩苷对体外培养的结肠癌细胞(CT-26,LoVo,Caco-2)的增殖具有明显的抑制作用,并呈剂量-效应关系,而对正常肾小球系膜细胞的毒性较小;明场和荧光图像显示汉黄芩苷作用后LoVo细胞出现明显的凋亡特征;流式细胞仪分析显示汉黄芩苷可以诱导LoVo细胞发生凋亡;RT-PCR检测结果显示汉黄芩苷可抑制LoVo细胞中bcl-2 mRNA的表达,促进bax mRNA的表达。结论汉黄芩苷可以显著抑制结肠癌细胞的增殖并诱导其凋亡,而对正常细胞影响较小,其机制可能与抗凋亡基因bcl-2表达下调,促凋亡基因bax表达上调有关。

Objective To determine the effect of Wogonoside on the proliferation of colon cancer cells and glomerular mesangial cells, and to investigate its mechanism of inducing apoptosis of tumor cells. Methods The colon cancer cells and glomerular mesangial cells cultured in vitro were treated with different concentrations of Wogonoside. MTT assay was used to examine the cell viability. The confocal laser scaning microscopy was used to de-termine the morphological changes of LoVo cells. The flow cytometry was used to test the apoptosis of LoVo cells. RT-PCR was used to determine the expression of apoptosis-related gene bax and bcl-2 in LoVo cells. Results The findings of MTT showed that Wogonoside could significantly inhibit the proliferation of the in vitro cultured colon cancer cells（CT-26, LoVo, Caco-2）, with a relationship of dose-effect, but had less toxicity on normal glomerular mesangial cells. The optical and fluorescent images showed that there were significant apoptosis characteristics in the LoVo cells after the Wogonoside treatment. The flow cytometry analysis showed that Wogonoside could induce apoptosis of LoVo cells. The findings of RT-PCR showed that Wogonoside could inhibit the mRNA expression of bcl-2 and promote the mRNA expression of bax. Conclusion Wogonoside can significantly inhibit the proliferation and induce apoptosis of colon cancer cells with less effect on normal cells. Its mechanism may be related to the down-regulation of anti-apoptotic gene bcl-2 and pro-apoptotic gene bax.