基础医学与临床 ›› 2016, Vol. 36 ›› Issue (2): 211-217.

• 研究论文 • 上一篇    下一篇

全反式维甲酸对小鼠肝癌细胞Hepa1-6增殖、迁移、侵袭的影响及其机制

崔洁洁1,龚梦嘉1,何昀2,毕杨1   

  1. 1. 重庆医科大学附属儿童医院儿研所干细胞实验室//儿童发育疾病研究教育部重点实验室//重庆市干细胞治疗工程技术研究中心
    2. 重庆医科大学
  • 收稿日期:2015-08-03 修回日期:2015-11-11 出版日期:2016-02-05 发布日期:2016-01-21
  • 通讯作者: 毕杨 E-mail:yangbi1981@cqmu.edu.cn
  • 基金资助:
    国家自然科学基金

Effect and mechanism of all-trans retinoic acid on proliferation, migration, and invasion of mouse hepatocellular carcinoma cell line Hepa1-6

  • Received:2015-08-03 Revised:2015-11-11 Online:2016-02-05 Published:2016-01-21
  • Contact: Yang Bi E-mail:yangbi1981@cqmu.edu.cn
  • Supported by:
    National Natural Science Foundation of China

摘要: 目的探讨不同浓度的全反式维甲酸(all-trans retinoic acid, ATRA)对小鼠肝癌细胞Hepa1-6体外增殖、凋亡、迁移和侵袭的影响及肝癌细胞间质标志蛋白和miR200家族的表达情况。方法 以Hepa1-6小鼠肝癌细胞为研究对象,给予0、0.1、1.0和10.0 μmol/L终浓度的ATRA处理,结晶紫染色检测细胞增殖,台盼蓝拒染实验计数活细胞。Hoechst检测细胞凋亡,划痕实验检测迁徙能力,Transwell实验检测侵袭能力,荧光定量 PCR(real-time PCR)法检测间质标志蛋白N-cadherin、sail和vimentin和0和10 μmol/L ATRA处理后的miR200家族的mRNA表达。结果 ATRA处理后Hepa1-6细胞的增殖、迁移、侵袭能力明显下降(P<0.05),凋亡率增高(P<0.05),间质标志蛋白N-cadherin、sail和vimentin的表达明显下调(P<0.05),ATRA的浓度越高,这些作用越明显。10 μmol/L ATRA处理后miRNA200a-3p,200c-3p,141-3p显著上调。结论 ATRA呈浓度依赖性促进肿瘤细胞Hepa1-6凋亡、抑制其增殖、迁移及侵袭能力,这可能与ATRA上调microRNA200家族,抑制细胞的间质表型有关。

关键词: 关键词:肝肿瘤细胞, 全反式维甲酸, 迁移, 侵袭, microRNA

Abstract: Objective To observe the effect of all-trans retinoic acid (ATRA) on the proliferation, apoptosis, migration and invasion of hepatocellular carcinoma cell line Hepa1-6 cell and investigate the mRNA expression of mesenchymal markers and microRNAs. Methods The Hepa1-6 cells were treated with DMEM containing different concentrations of ATRA (0, 0.1, 1.0, 10.0 μmol/L). Trypan blue and Crystal Violet Staining were used to assess cell proliferation at indicated time points. The apoptosis was examined by Hoechst Staining. Migration and invasion abilities of cells were detected by Wound Healing assay and Transwell assay, respectively. The mRNA expression of mesenchymal markers (N-cadherin, sail, vimentin) of Hepa1-6 with ATRA at different concentrations and miRNA200s with10.0 μmol/L ATRA treatment were detected by real-time PCR. Results Compared with control group (0 μmol/L), after treatment with ATRA, the proliferation, migration and invasion capacity of Hepa1-6 cells were obviously inhibited (P<0.05), and the apoptosis rates increased significantly. The mRNA expression of mesenchymal markers (N-cadherin, sail, vimentin) was decreased. Furthermore, the role of ATRA on these was strengthened with the increased concentrations of ATRA treatment. In addition, miR200a-3p, miR200c-3p, miR141-3p were up-regulated. Conclusion ATRA can inhibit the growth, migration, invasion and down-regulate mesenchymal markers (N-cadherin, sail, vimentin) of hepatocellular carcinoma cell line Hepa1-6 cell, induce its apoptosis in a concentration-dependent manner. These may be related to the up-regulation of miR200s and inhibition of mesenchymal phenotypes.

Key words: Key words: hepatocellular carcinoma cells, all-trans retinoic, migration, invasion, microRNA

中图分类号: