重组豹蛙抗瘤酶促进大鼠肝癌RH-35细胞凋亡  

Recombinant onconase promotes the apoptosis of rat hepatoma cell RH-35

作　　者：许晓亚[1,2] 靳伟[1,2] 杨刚刚[1,2] 张全义 吕中原 秦艳丽[1,2] 徐存拴 

机构地区：[1]河南师范大学生命科学学院,河南新乡453007 [2]河南省-科技部共建细胞分化国家重点实验室培育基地和河南省生物工程重点实验室,河南新乡453007 [3]河南新乡华星药厂,河南新乡453731

出　　处：《基础医学与临床》2016年第1期1-5,共5页Basic and Clinical Medicine

基　　金：国家973前期研究专项(2012CB722304);河南省自然科学基金(142300413227);河南省高等学校重点科研项目(15A180042);河南省重大科技攻关项目(111100910600)

摘　　要：目的研究重组豹蛙抗瘤酶(r ONC)对大鼠肝癌RH-35细胞增殖和凋亡的作用。方法用不同浓度的r ONC处理体外培养的RH-35细胞,用SRB法检测细胞增殖;Hoechst染色法测定细胞活性;用流式细胞术检测细胞周期;用qRT-PCR和Western blot法检测相关基因和蛋白的变化。结果 r ONC作用于RH-35细胞24 h后呈时间和浓度依赖性,显著抑制增殖(P<0.05),IC_(50)为7.38μmol/L。rONC作用于RH-35细胞后,细胞核出现了显著的凋亡特征。G0/G1期的细胞比例显著上升(P<0.05),增殖能力减弱。r ONC作用于RH-35细胞后,BAX mRNA和蛋白水平升高(P<0.05),而PCNA,CCNA2和BCL-2 mRNA和蛋白水平降低(P<0.05)。结论 r ONC可能通过上调BAX表达和下调PCNA,CCNA2和BCL-2表达,促进RH-35细胞凋亡,抑制细胞增殖。Objective To investigate the apoptosis and cell growth inhibition by recombinant onconase（ rONC） on rat hepatoma cell RH-35. Methods After RH-35 cells were treated in vitro with various concentrations of rONC.Cel1 proliferation was determined by the SRB method and the bioactivities of these cell were detected by Hoechst33258. The effect of rONC on cell cycle was detected by flow cytometry assay. The expression of cell proliferation related genes and proteins after RH-35 cell treatment with rONC were detected by qRT-PCR and Western blot. Results The proliferation of RH-35 cell was significantly inhibited after treatment with rONC for 24 hours and this inhibitory action was positively correlated with time and dosage（ P〈0. 05）. The value of IC50was7. 38 μmol / L. The nucleus showed some characteristic of apoptosis after RH-35 cell treatment with rONC. The proportion of cells in G0/ G1 phases remarkably raised after 24 h treatment with rONC,rONC down-regulated the expres-sion of PCNA,CCNA2,BCL-2 and up-regulated BAX gene expression and protein synthesis. Conclusions rONC may inhibit the growth and induce apoptosis of RH-35 cell in vitro due to down-regulate the expression of PCNA,CCNA2,BCL-2 and up-regulate expression of BAX.

关 键 词：重组豹蛙抗瘤酶 大鼠肝癌细胞 生长抑制 细胞凋亡 

分 类 号：R735.7[医药卫生—肿瘤]