解剖学报 ›› 2015, Vol. 46 ›› Issue (6): 780-784.doi: 10.16098/j.issn.0529-1356.2015.06.009

• 肿瘤生物学 • 上一篇    下一篇

氧化损伤对骨髓源性肝前体细胞恶性转化的影响

文朝阳1 史争鸣2 赵娜3 薛晓伟3 丰平3 王学江3*   

  1. 1. 首都医科大学医学生物学实验教学中心,北京 100069; 2. 北京积水潭医院外科,北京 100035; 3. 首都医科大学生理学与病理生理学系,北京 100069
  • 收稿日期:2015-06-17 修回日期:2015-08-25 出版日期:2015-12-06 发布日期:2015-12-06
  • 通讯作者: 王学江 E-mail:wangwenwen1722@163.com
  • 基金资助:

    尾加压素及其受体介导卵圆细胞在肝癌形成中的作用和分子机制;北京市属高等学校创新团队建设与教师职业发展计划项目

Effect of oxidative damage on malignant transformation of bone marrow-derived hepatic progenitor cells

WEN Zhao-yang1 SHI Zheng-ming2 ZHAO Na3 XUE Xiao-wei3 FENG Ping3 WANG Xue-jiang 3*   

  1. 1.Medical Biology Experimental Teaching Center of Capital Medical University, Beijing 100069, China; 2.Surgry of Beijing Jishuitan Hospital, Beijing 100035, China; 3. Physiological and Pathophysiological Department of Capital Medical University, Beijing 100069, China
  • Received:2015-06-17 Revised:2015-08-25 Online:2015-12-06 Published:2015-12-06
  • Contact: WANG Xue-jiang E-mail:wangwenwen1722@163.com

摘要:

目的 探讨活性氧自由基对骨髓源性肝前体细胞恶性转化的影响。 方法 取经过鉴定的3代骨髓间充质干细胞(BMSCs)分为3组:对照组(BMSCs),生长因子诱导组(BMSCs+HGF+EGF),过氧化氢刺激组(BMSCs+HGF+EGF+ H2O2)。分别在细胞培养的第7天、14天、21天、28天用RT-PCR方法检测各组细胞的白蛋白(ALB)、甲胎蛋白(AFP)、角蛋白18 (CK18) mRNA的表达。用免疫细胞化学方法检测细胞γ-谷氨酰转酞酶(γ-GT)的表达,用Schiff染色方法检测细胞DNA含量,用流式细胞仪检测非整倍体细胞。 结果 生长因子诱导组及过氧化氢刺激组细胞有ALB mRNA、AFP mRNA及CK18 mRNA的表达,过氧化氢刺激组细胞AFP mRNA、CK18 mRNA、γ-GT的表达、DNA含量及非整倍体细胞均高于生长因子诱导组(P<0.05)。 结论 骨髓间充质干细胞在体外诱导成为肝前体细胞,活性氧自由基可促使肝前体细胞过度增殖并恶性转化。

关键词: 骨髓间充质干细胞, 氧化损伤, 恶性分化, 肝癌, 微环境, 反转录-聚合酶链反应, 大鼠

Abstract:

Objective To explore the role of oxidative damage in malignant transformation of bone marrow-derived liver stem cells. Methods Bone marrow mesenchymal stem cells (BMSCs) were divided into three groups: control group(BMSCs), GF induction group(BMSCs+GF)and Hydrogen peroxide stimulate group(BMSCs+GF+ H2O2). Albumin (ALB) mRNA, alpha-fetoprotein (AFP) mRNA and CK18 mRNA were detected by RT-PCR at 7d, 14d, 21d and 28d. Gamma glutamyl transpeptidase (γ-GT) was detected by immunocytochemical technique. DNA contents were detected by Schiff dyeing. Aneuploid somatic cells were detected by flow cytometry. Results The results of in vitro experiments showed that there were expressions of ALB mRNA, AFP mRNA and CK18mRNA in group BMSCs+GF and group BMSCs+GF+ H2O2. The expressions of AFP mRNA, CK18mRNA, γ-GT, DNA contents and aneuploid somatic cells in group BMSCs+GF+ H2O2 were higher than that in group BMSCs+GF. Conclusion Reactive oxygen species can induce bone marrow mesenchymal stem cell proliferation and malignant transformation.

Key words: Bone marrow mesenchymal stem cell, Oxidative damage, Malignant transformation, Liver cancer, Microenvironment, Reverse transcription-polymerase chain reaction, Rat