摘要
目的探讨苦参碱对结肠癌耐药细胞化疗药物敏感性及自噬水平的影响。方法分别采用浓度为0.5mg/ml、1 mg/ml、2 mg/ml、4 mg/ml、8 mg/ml苦参碱作用于结肠癌细胞HCT-8/VCR,分别在24 h、48 h,采用CCK-8法检测苦参碱对结肠癌细胞HCT-8及其耐药细胞HCT-8/VCR的抑制作用,并计算出抑制率;运用Western Blotting检测自噬相关基因P62、Beclin-1、LC3蛋白表达。采用流式细胞术检测苦参碱作用耐药细胞后的细胞凋亡率。结果在苦参碱作用24 h和48 h后,HCT-8细胞增殖的抑制作用随着苦参碱浓度的增加而增强,苦参碱对HCT-8细胞增殖的抑制作用呈浓度依赖性,差异有统计学意义(P<0.001)。对各组细胞的凋亡率检测发现,采用苦参碱的实验组的细胞凋亡率大大高于对照组,差异有统计学意义(P<0.001)。在苦参碱应用24 h后,HCT-8/VCR细胞质中出现了自噬泡。Western Blotting检测显示随着苦参碱浓度的增加,P62、Beclin-1、LC3蛋白的表达水平也随之增强。结论苦参碱能够提高结肠癌耐药细胞的自噬水平,逆转结肠癌耐药细胞HCT-8/VCR的多药耐药性,诱导细胞凋亡,抑制细胞增殖。
Objective To explore effect of matrine on chemotherapeutic drug sensitivity and autophagy levels in colon cancer resistant cells.Methods Matrine with concentrations of 0.5 mg/ml, 1 mg/ml, 2 mg/ml,4 mg/ml and 8 mg/ml were respectively used in colon cancer cell HCT-8/VCR.24 hours and 48 hours after application of matrine,the inhibitory effects of matrine on colon cancer cell HCT-8 and its drug resistance cell HCT-8/VCR were detected by CCK-8,and inhibition rate was calculated.In addition,Western Blotting was used to detect the expressions of autophagy related genes including P62, Beclin- 1 and LC3,and flow cytometry was used to detect apoptosis rate after matrine affecting on resistant cells.Results 24 hours and 48 hours after application of matrine ,the inhibition of HCT-8 cell proliferation enhanced with the increase of matrine concentration, inhibition effect of matrine on HCT-8 cell proliferation showed a concentration dependent manner, so difference was statistically significant （P〈0.001）.Detection of apoptosis rate in each group showed that the apoptosis rate of the experimental group（ given matrine） was significantly higher than that of the control group, so difference was statistically significant （P〈 0.001 ）.24 hours after the application of matrine ,autophagic vacuoles were found in HCT-8/VCR cytoplasm.Blotting Western detection showed that the expression levels of P62, Beclin-1 and LC3 increased with the increase of matrine concentration.Conclusion Matrine can increase the levels of autophagy in colon cancer resistant cells,reverse muhidrug resistance of colon cancer cell HCT-8/VCR,induce cell apoptosis and inhibit cell proliferation.
出处
《右江医学》
2016年第6期610-613,共4页
Chinese Youjiang Medical Journal
基金
广西高校重点实验室开放课题(NO:kfkt2016010)
2016年广西研究生教育创新计划项目(20161007)
