积雪草酸对人舌癌TCA-8113细胞增殖、凋亡的影响  

Inhibitory effect of asiatic acid on proliferation and apoptosis of TCA-8113 human tongue carclinoma cells

作　　者：董易 褚冬青 马春宇 黄建华 王一 DONG Yi;CHU Dong-qing;MA Chun-yu;HUANG Jian-hua;WANG Yi(Clinical Lab, the First Affiliated Hospitalof Jinzhou Medical University, Jinzhou, Liaoning 121000,China;Key Lab of Surgery, the First Affiliated Hospitalof Jinzhou Medical University, Jinzhou, Liaoning 121000,China)

机构地区：[1]锦州医科大学附属第一医院检验科,辽宁锦州121000 [2]锦州医科大学附属第一医院外科重点实验室,辽宁锦州121000

出　　处：《中国药理学通报》2019年第11期1596-1601,共6页Chinese Pharmacological Bulletin

基　　金：辽宁省教育厅重点实验室建设项目(No LZ2015051);辽宁省自然科学基金项目(No 20170540368)

摘　　要：目的 探究积雪草酸(asiatic acid,AA)体外对人舌癌TCA-8113细胞增殖、凋亡、细胞周期阻滞的作用及其可能的作用机制。方法 MTT法检测不同浓度的AA(0、20、30、40、50 μmol·L -1 )作用于TCA-8113细胞24 h的增殖抑制作用;集落形成实验检测AA对TCA-8113细胞集落形成率的影响;Hoechst 33342染色法、Annexin V-FITC/PI法检测AA对TCA-8113细胞凋亡的影响;流式细胞术分析细胞周期变化;Western blot检测AA对TCA-8113细胞中Bcl-2、Bax、cleaved caspase-3及p53、p21蛋白表达的影响。结果 AA对TCA-8113细胞抑制率呈浓度依赖性( P <0.05),其IC 50 值为42.13 μmol·L -1 ,细胞集落形成能力降低;随AA浓度的增大,细胞凋亡率增加( P <0.05);当AA浓度达到30 μmol·L -1 时,细胞周期逐渐阻滞在G 2/M期;Western blot显示,p53、p21及Bax表达升高,Bcl-2表达降低( P <0.05),呈浓度依赖性。结论 AA明显抑制TCA-8113细胞的增殖,诱导其凋亡,其机制可能与调控p53通路相关蛋白p53、p21、Bax,以及抑制Bcl-2的表达有关,同时AA能使TCA-8113细胞周期阻滞在G 2/M期。Aim To investigate the effect of asiatic acid(AA) on the proliferation, apoptosis and cell cycle arrest of human tongue carcinoma TCA-8113 cells, and the possible mechanism. Methods The inhibitory effect of AA(0, 20, 30, 40, 50 μmol·L -1 ) at different concentrations on TCA-8113 cells for 24 h was determined by MTT. The effect of AA on the colony formation rate of TCA-8113 cells was detected by colony formation test;The apoptosis of TCA-8113 cells by AA was detected by Hoechst 33342 staining and Annexin V-FITC/PI staining. Cell cycle changes were analyzed by flow cytometry. The expressions of protein in human tongue cancer cells, such as bcl-2, Bax, cleaved caspase-3, p53 and p21 proteins were detected by Western blot. Results The inhibitory rate of AA on TCA-8113 cells was concentration-dependent( P <0.05), with the IC 50 concentration of 42.13 μmol·L -1 , and the ability of cell colony formation decreased. The apoptosis rate increased with the increase of AA concentration( P <0.05). The concentration of AA 30 μmol·L -1 gradually blocked the cell cycle in G 2/M phase. Western blot analysis showed that the expressions of p53, p21 and Bax increased, while Bcl-2 decreased in a dose-dependent effect( P <0.05). Conclusions AA can significantly inhibit the proliferation and apoptosis of human tongue carcinoma TCA-8113 cells, and its mechanism may be related to the regulation of p53 pathway related proteins p53, p21, Bax and the inhibition of Bcl-2 expression, and AA can arrest TCA-8113 cells cycle in G 2/M phase.

关 键 词：积雪草酸 舌癌 TCA-8113 凋亡 BCL-2 cleaved CASPASE-3 P53 

分 类 号：R282.71[医药卫生—中药学] R329.24[医药卫生—中医学]