拟康氏木霉胞外多糖对人结肠癌细胞HCT116增殖及凋亡的影响  

Effect of exopolysaccharide from Trichoderma pseudokoningii on proliferation and apoptosis of human colon cancer cell line HCT116

作　　者：李萍 沈学彬 周玉燕 李艳 高娃 陈靠山 王国栋 LI Ping;SHEN Xue-bin;ZHOU Yu-yan;LI Yan;GAO Wa;CHEN Kao-shan;WANG Guo-dong(School of Pharmacy, Wannan Medical College;Anhui Provincial Engineering Research Center for Polysaccharide Drugs;Anhui Provincial Key Lab of Active Biological Macro-molecules, Wuhu, Anhui 241002, China)

机构地区：[1]皖南医学院药学院 [2]安徽省多糖药物工程技术研究中心 [3]活性生物大分子研究安徽省重点实验室,安徽芜湖241002

出　　处：《中国药理学通报》2019年第11期1621-1626,共6页Chinese Pharmacological Bulletin

基　　金：安徽省高校自然科学研究项目(No KJ2018ZD025,KJ2017A257);安徽省自然科学基金资助项目(No 1908085MH248);皖南医学院大学生科研资助金项目(No WK2018S36,WK2018S37)

摘　　要：目的 探讨拟康氏木霉胞外多糖(exopolysaccharide from Trichoderma pseudokoningii ,EPS)对人结肠癌细胞株HCT116细胞增殖和凋亡的影响。方法 CCK-8法和克隆形成实验检测EPS对HCT116细胞增殖的影响,流式细胞术检测细胞凋亡率,JC-1检测线粒体膜电位,Western blot检测蛋白Bcl-2、Bax的表达,试剂盒检测caspase-9、caspase-8、caspase-3活性。结果 EPS在0~800 mg·L -1 范围内可抑制HCT116细胞增殖,并具有时间与浓度依赖性。EPS处理组HCT116克隆形成能力减弱,线粒体膜电位下降,Bcl-2/Bax比值明显下降,caspase-9、caspase-8和caspase-3活性均明显增高。结论 EPS可以明显抑制人结肠癌细胞株HCT116增殖,并诱导其凋亡。Aim To study the effect of exopolysaccharide from Trichoderma pseudokoningii (EPS) on the proliferation and apoptosis of human colon cancer cell line HCT116. Methods The proliferation of HCT116 cells treated with EPS was examined by CCK-8 assay. The effect of EPS on the clone formation of HCT116 cells was detected by crystal violet staining. Cell apoptotic rate was determined by flow cytometry. The changes of mitochondrial membrane potential were observed by JC-1. The expressions of apoptosis-related proteins Bcl-2 and Bax were analyzed by Western blot, and caspase-9, caspase-8 and caspase-3 expressions were detected by the kit. Results EPS dose- and time-dependently inhibited the proliferation of the HCT116 cells in the range of 0~800 mg·L -1 . With increase of EPS concentration, the colony-formation ability of HCT116 cells decreased;the proportion of apoptotic cells increased and mitochondrial membrane potential decreased;the expression of Bcl-2 protein decreased, while the expression of Bax protein increased;the ratio of Bcl-2/Bax decreased gradually;caspase-9, caspase-8 and caspase-3 activities significantly increased. Conclusions EPS can inhibit proliferation of HCT116 and induce its apoptosis by up-regulating expression of Bax, caspase-9 ,caspase-8, caspase-3 and down-regulating the expression of Bcl-2.

关 键 词：胞外多糖 拟康氏木霉 信号通路 细胞凋亡 增殖 结肠癌 

分 类 号：R285.5[医药卫生—中药学] R329.24[医药卫生—中医学]