基础医学与临床 ›› 2016, Vol. 36 ›› Issue (1): 1-5.

• 研究论文 •    下一篇

重组豹蛙抗瘤酶促进大鼠肝癌RH-35细胞凋亡

许晓亚1,靳伟1,杨刚刚1,张全义2,吕中原2,秦艳丽1,徐存拴1   

  1. 1. 河南师范大学
    2. 河南新乡华星药厂
  • 收稿日期:2015-07-08 修回日期:2015-10-08 出版日期:2016-01-05 发布日期:2015-12-29
  • 通讯作者: 徐存拴 E-mail:cellkeylab@126.com
  • 基金资助:
    国家973前期研究专项;河南省自然科学基金;河南省高等学校重点科研项目;河南省重大科技攻关项目

Recombination Onconase promotes the apoptosis of rat hepatoma cell RH-35

  • Received:2015-07-08 Revised:2015-10-08 Online:2016-01-05 Published:2015-12-29

摘要: 目的 研究重组豹蛙抗瘤酶(rONC)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rONC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 rONC作用于RH-35细胞24h后呈时间和浓度依赖性,显著抑制增殖(P<0.05),IC50为7.38?mol/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。rONC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:rONC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。

关键词: 关键词:重组豹蛙抗瘤酶, 大鼠肝癌细胞, 生长抑制, 细胞凋亡

Abstract: Objective To investigate the apoptosis and cell growth inhibition in vitro effects of recombination onconase (rONC) on rat hepatoma cell RH-35. Methods After RH-35 cell in vitro were treated with various concentrations of rONC. Cel1 proliferation were determined by the SRB method and the bioactivities of these cell were detected by Hoechst33258. The effect of rONC on cell cycle were detected by flow cytometry assay. The expression levels of cell proliferation related genes and proteins after RH-35 cell treated with rONC were detected by qRT-PCR and Western blot. Results The proliferation of RH-35 cell were significantly inhibited after treated with rONC for 24 hours and this inhibitory action was positive correlated with time and dose-dependent (P<0.05) . The value of IC50 was 7.38 ?mol/L. The nucleus appeared some characteristic of apoptosis after RH-35 cell treated with rONC .The proportion of cells in G0/G1 phases remarkably rise after 24 h treatment with rONC, rONC could down-regulate the expression level of PCNA, CCNA2, BCL-2 and up-regulate BAX gene levels and protein levels. Conclusion rONC could inhibit the growth and induce apoptosis of RH-35 cell in vitro due to the down-regulated expression of PCNA, CCNA2, BCL-2 and up-regulated expression of BAX.

Key words: Key words: recombination onconase, rat hepatoma cell line RH-35, growth inhibition, apoptosis

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