Abstract：Objective To investigate the mechanism of extracellular inhibition of GBC-SD cell proliferation,invasion and apoptosis by Cinobufotalin in gallbladder carcinoma.Methods The proliferation of GBC-SD cells was determined by CCK-8 cell counting and plate clonogenesis assay after treatment with different concentrations of Cinobufotalin(0,100,300,500 nmol/L).AnnexinⅤ-FITC/PI double staining method was used to detect the effect of different concentrations of Cinobufotalin(0,100,300 and 500 nmol/L)on apoptosis of GBC-SD cells.Flow cytometry(FCM)was used to detect the effect of Cinobufotalin on GBC-SD cell cycle.The change of invasion ability of gallbladder carcinoma cells treated with Cinobufotalin was observed by Transwell chamber experiment.Among them,the concentration of Cinobufotalin of 0 nmol/L was used as the blank drug-free group.Results Compared with the blank drug-free group,Cinobufotalin inhibited the proliferation of GBC-SD cells in a time-and dose-dependent manner(P＜0.01).Annexin V-FITC/PI double staining showed that Cinobufotalin had the ability to induce apoptosis of GBC-SD cells (P＜0.01).With the increase of the concentration of Cinobufotalin,the survival rate of GBC-SD cells decreased significantly,and the apoptosis rates (early apoptosis rate and late apoptosis rate)increased significantly (P＜0.01).Cinobufotalin could promote the expression levels of pro-apoptotic proteins Cleaved-PARP,Cleaved-caspase3,Cleaved-caspase9,BAX.And Cinobufotalin could decreased the expression of anti-apoptotic protein Bcl-2.The proportions of cells distributed in G2/M phase increased with the increase of the concentration of Cinobufotalin,and the differences were statistically significant(P＜0.05).The expression levels of CDK1 and Cyclin B1 in G2/M progressiverelated proteins decreased with the increase of the concentration of Cinobufotalin (P＜0.01).The number of invasive cells was decreased with the increase of the concentration of Cinobufotalin (P＜0.001).The expression levels of E-cadherin,a protein associated with invasive inhibition,were higher than those of blank drug-free group,and the expression levels of N-cadherin and Vimentin,which were protein associated with invasive inhibition,were lower than those of blank drug-free group,with statistical significances(P＜0.01).Conclusion Cinobufotalin can effectively inhibit the proliferation and invasion of GBC-SD cells in gallbladder carcinoma and induce its apoptosis.
李俊杰；邬海峰；张东明；张志平；李明章. 华蟾素体外抑制胆囊癌G BC-SD细胞增殖与侵袭的机制研究[J]. 中国当代医药, 2021, 28(31): 27-32.
LI Jun-jie；WU Hai-feng；ZHANG Dong-ming；ZHANG Zhi-ping；LI Ming-zhang. Study on the mechanism of extracellular inhibition of GBC-SD cell proliferation and invasion by Cinobufotalin in gallbladder carcinoma. 中国当代医药, 2021, 28(31): 27-32.
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