鸢尾黄酮通过促进自噬抑制结肠癌细胞增殖并诱导其凋亡研究  

Study of Tectorigenin Inhibits the Proliferation and Induces Apoptosis of Colon Cancer Cells by Promoting Autophagy

作　　者：洪晓泉[1] 傅锦波[1] 林福生[1] 罗晔哲[1] 林恩德[1] HONG Xiao-quan;FU Jin-bo;LIN Fu-sheng;LUO Ye-zhe;LIN En-de(Department of General Surgery,Zhongshan Hospital Affiliated to Xiamen University,Fujian,Xiamen361004)

机构地区：[1]厦门大学附属中山医院普通外科,福建厦门361004

出　　处：《实用中西医结合临床》2021年第21期4-8,共5页Practical Clinical Journal of Integrated Traditional Chinese and Western Medicine

基　　金：福建省自然科学基金项目(编号:2016J01617)。

摘　　要：目的:探究鸢尾黄酮对结肠癌细胞自噬及其对细胞增殖能力和凋亡的影响。方法:鸢尾黄酮对结肠癌细胞的半抑制浓度(IC50)采用CCK-8法检测;结肠癌细胞的长期增殖能力采用克隆平板实验检测;采用CCK-8法检测不同抑制剂预处理后鸢尾黄酮对结肠癌细胞增殖活性的影响;采用蛋白免疫印迹法检测自噬相关蛋白的变化情况;透射电镜观察结肠癌细胞中自噬体的数量;采用CCK-8法检测小干扰RNA(siRNA)沉默自噬关键蛋白ATG5转染后鸢尾黄酮对结肠癌细胞增殖活性的影响。结果:通过不同浓度鸢尾黄酮作用下结肠癌细胞系SW480的生长抑制情况绘制IC50曲线,确定其IC50为(221.90±35.31)μM;克隆平板实验表明鸢尾黄酮对结肠癌细胞的长期增殖能力具有抑制作用;凋亡抑制剂Z-VAD显著逆转了鸢尾黄酮对结肠癌细胞的增殖抑制作用;Western blot实验结果表明,随着鸢尾黄酮作用浓度的升高,SW480细胞内LC3-Ⅱ含量逐渐上升,p62含量下降;电镜结果表明,经鸢尾黄酮作用后SW480细胞内自噬体含量明显增加;沉默ATG5可部分逆转鸢尾黄酮对SW480细胞的增殖抑制作用。结论:鸢尾黄酮通过促进自噬抑制了结肠癌细胞的增殖能力,并诱导结肠癌细胞的凋亡。Objective:To investigate the influence of tectorigenin on autophagy,proliferation ability and apoptosis of colon cancer cells.Methods:The half inhibitory concentration(IC50)of tectorigenin on colon cancer cells was detected by CCK-8 method;the long-term proliferation ability of colon cancer cells was detected by clone plate test;CCK-8 method was used to detect the effect of tectorigenin pretreated with different inhibitors on the proliferation of colon cancer cells;the changes of autophagy related proteins were detected by Western Blot method;the number of autophagosomes in colon cancer cells was observed by transmission electron microscope;CCK-8 method was used to detect the effect of tectorigenin on the proliferation of colon cancer cells after small interfering RNA(siRNA)silencing autophagy key protein ATG5 transfection.Results:The IC50 curve of colon cancer cell line SW480 was drawn by the growth inhibition of tectorigenin at different concentrations,and the IC50 was determined to be(221.90±35.31)μM;cloning plate experiment showed that tectorigenin could inhibit the long-term proliferation of colon cancer cells;apoptosis inhibitor Z-VAD significantly reversed the inhibitory effect of tectorigenin on the proliferation of colon cancer cells;the experimental results of Western Blot showed that the content of LC3-Ⅱin SW480 cells gradually increased and the content of p62 decreased with the increase of the concentration of tectorigenin;the results of electron microscopy showed that the content of autophagosome in SW480 cells increased significantly after treated with tectorigenin;silencing ATG5 could partially reverse the inhibitory effect of tectorigenin on the proliferation of SW480 cells.Conclusion:Tectorigenin can inhibit the proliferation of colon cancer cells and induces apoptosis of colon cancer cells by promoting autophagy.

关 键 词：结肠癌 鸢尾黄酮 自噬 增殖 凋亡 

分 类 号：R285.5[医药卫生—中药学]