黄芩苷诱导人结肠癌细胞周期阻滞和凋亡的体内外研究

发布时间：2018-03-11 04:02

本文选题：结肠肿瘤　切入点：黄芩苷　出处：《肿瘤》2017年03期 　论文类型：期刊论文

【摘要】：目的:探讨黄芩苷在体内外对结肠癌细胞凋亡和细胞周期的影响,以及其可能的分子作用机制。方法:不同质量浓度(0、50、100、200、400和800μg/mL)的黄芩苷分别作用人正常结直肠黏膜FHC细胞和人结肠癌HCT116细胞48 h后,倒置显微镜下观察细胞形态变化,并采用MTT法检测各组细胞活力变化。FCM法检测黄芩苷干预后结肠癌HCT116细胞凋亡率和细胞周期分布的变化;同时,采用蛋白质印迹法检测凋亡相关蛋白聚腺苷二磷酸核糖聚合酶-1(poly ADP-ribose polymerase-1,Parp-1)、X连锁凋亡抑制蛋白(X-linked inhibitor of apoptosis protein,XIAP)、核因子-κB(nuclear factor-κB,NF-κB)、caspase 3、p53、Bcl-2和Bax,以及细胞周期相关蛋白cyclin D1和cyclin B1表达水平的变化。此外,通过构建结直肠癌HCT116细胞的原位移植瘤小鼠模型,观察黄芩苷灌胃治疗后小鼠体质量以及体内肿瘤生长体积的变化,并应用TUNEL法检测黄芩苷对移植瘤小鼠体内肿瘤细胞凋亡的影响。结果:与黄芩苷未处理的对照组相比,50~800μg/mL黄芩苷对结肠癌HCT116细胞活力有明显的抑制作用(P值均0.05)。而且黄芩苷对正常结直肠黏膜细胞FHC的半数抑制浓度(half maximal inhibitory concentration,IC50)值明显高于HCT116细胞(P0.01)。200和400μg/mL黄芩苷处理后,结肠癌HCT116细胞的凋亡率明显升高(P值均0.01);50和100μg/mL黄芩苷作用48 h后,Parp-1和caspase 3剪切体蛋白的表达水平比对照组均明显升高(P值均0.01),而XIAP、NF-κB和Bcl-2蛋白的表达水平均明显降低(P值均0.05)。100和200μg/mL黄芩苷处理后,HCT116细胞周期阻滞在G1期(P值均0.01);50和100μg/mL黄芩苷作用48 h后,cyclin D1和cyclin B1蛋白的表达水平均明显降低(P值均0.01)。黄芩苷可以抑制小鼠原位移植瘤的生长(P0.01),并促进肿瘤细胞凋亡,但是对小鼠体质量无明显影响(P0.05)。结论:黄芩苷在体内外均可以明显抑制结肠癌HCT116细胞的生长活性,诱导细胞凋亡,并使细胞周期阻滞于G1期。
[Abstract]:Objective: to investigate the effects of baicalin on apoptosis and cell cycle of colon cancer cells in vitro and in vivo. Methods: baicalin at different concentrations (100 渭 g / mL and 800 渭 g / mL) treated FHC cells of normal colorectal mucosa and HCT116 cells of colon cancer respectively for 48 h, and observed the morphologic changes of cells under inverted microscope. The apoptosis rate and cell cycle distribution of colon cancer HCT116 cells after baicalin intervention were detected by MTT assay. The expression of apoptosis associated protein polyadenosine diphosphate polymerase 1 (poly ADP-ribose polymerase 1) and cyclin cyclin D1 and cyclin B1 were detected by Western blotting. The expression levels of X-linked inhibitor of apoptosis protein XIAPP, nuclear factor- 魏 BNF- 魏 Bncaspase 3p53Bcl-2 and cyclin B1, and cyclin-associated protein cyclin D1 and cyclin B1 were detected by Western blotting. In order to observe the changes of body mass and tumor growth volume in mice after baicalin administration, the orthotopic transplanted tumor model of colorectal cancer HCT116 cells was established. The effect of baicalin on tumor cell apoptosis in transplanted tumor mice was detected by TUNEL method. Results: compared with the untreated control group, baicalin (50 ~ 800 渭 g / mL) significantly inhibited the viability of colon cancer HCT116 cells (P < 0.05). The half maximal inhibitory concentration (IC50) of baicalin on FHC of normal colorectal mucosal cells was significantly higher than that of HCT116 cells treated with baicalin at the concentration of 0.01 渭 g / mL and 400 渭 g / mL, respectively. The apoptotic rate of colon cancer HCT116 cells was significantly increased by 0.01g / mL baicalin 50 and 100 渭 g / mL baicalin for 48 h. After 48 h treatment, the expression levels of Shear body proteins of PIP 1 and caspase 3 were significantly higher than those of the control group, while the expression levels of XIAP NF- 魏 B and Bcl-2 protein were significantly decreased compared with those of the control group. The cell cycle arrest of HCT116 cells treated with baicalin (0.05 渭 g 路100) and 200 渭 g / mL baicalin was inhibited in G1 phase. The expression levels of cyclin D1 and cyclin B1 protein were significantly decreased after exposed to baicalin at G1 phase for 48 h. Baicalin could inhibit the proto-displacement of HCT116 cells. The expression of cyclin D1 and cyclin B1 protein was significantly decreased after treatment with baicalin at G _ 1 phase. Baicalin could inhibit the proto-displacement of HCT116 cells. The growth of tumor was P0.01 and the apoptosis of tumor cells was promoted. Conclusion: baicalin can significantly inhibit the growth activity of colon cancer HCT116 cells in vitro and in vivo, induce apoptosis and arrest cell cycle in G1 phase.
【作者单位】：南京师范大学生命科学学院江苏省分子与医学生物技术重点实验室;沈阳化工大学制药与生物工程学院生物工程教研室;
【分类号】：R735.35

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