高良姜总黄酮对宫颈癌细胞生长和肿瘤干细胞标记物表达水平的影响及意义

Effect of Galangal Total Flavonoids on the Growth of Cervical Carci-noma Cells and the Expression of Cancer Stem Cell Markers

目的：前期研究中，我们报道高良姜根来源的总黄酮（ general flavonoids， GFs）可抑制SiHa宫颈癌细胞增殖，诱导细胞凋亡。以往研究证明肿瘤干细胞是SiHa细胞群体的重要组成部分，故本研究探讨高良姜GFs影响细胞凋亡、细胞周期相关基因及肿瘤干细胞标记物表达调控的机理，为抗肿瘤药物研发提供依据。方法以0、100和200μg／ml剂量的高良姜GFs干预SiHa宫颈癌细胞24 h，提取细胞总RNA，采用荧光定量RT-PCR方法对CDK4、 bcl-2、 ALDH1 A1、 OCT4和Twist1等5种基因表达水平进行定量分析。结果高良姜GFs干预SiHa宫颈癌细胞后， CDK4和bcl-2基因表达水平显著下降（ P＜0.05），呈剂量依赖性，说明该GFs可能调节细胞增殖和生存相关的基因表达调控，诱导细胞凋亡；随着药物剂量增加，肿瘤干细胞标记物ALDH1 A1、 OCT4和Twist1表达水平也梯度性下降，并有统计学差异（ P＜0.05）。结论本研究结果提示高良姜GFs可能抑制肿瘤干细胞基因表达水平，从而抑制细胞增殖，促进细胞凋亡，此为揭示宫颈癌发病机制及抗肿瘤新药研发提供重要依据。

Objective We reported previously that total flavonoids （ TFs） isolated from galan-gal root inhibited the proliferation and induced apoptosis of SiHa cervical carcinoma cells.Cancer stem cells are demonstrated to be an important constituent of SiHa cells.Here, we investigated the mechanism by which galangal TFs influence the expression of genes related to cell apoptosis and cell cycle , and cancer stem cell markers.Methods SiHa cervical carcinoma cells were treated with ga-langal TFs at doses of 0, 100 and 200 μg/mL for 24 h.After isolation of cellular total RNA, the expression of five genes including CDK4, bcl-2, ALDH1 A1, OCT4 and Twist1 was analyzed by fluorescent quantitative RT-PCR.Results The expression of CDK4和bcl-2 genes in SiHa cervical carcinoma cells was significantly reduced in response to treatment with galangal TFs in a dose de-pendent manner （ P〈0.05 ） , indicating that the galangal TFs may regulate the expression of genes related to cell proliferation and cell survival.Meanwhile, treatment with galangal TFs reduced the＆amp;nbsp;expression of cancer stem cell markers ALDH1 A1, OCT4 and Twist1, with statistical significance found when compared to controls （ P〈0.05 ） .Conclusion Galangal TFs may inhibit the expres-sion of cancer stem cell markers, and then suppress cell proliferation and induce cell apoptosis of cancer cells, which provides evidence for study of mechanisms of cervical cancer development and the discovery of novel anticancer drugs.