中国癌症杂志 ›› 2015, Vol. 25 ›› Issue (7): 516-521.doi: 10.3969/j.issn.1007-3969.2015.07.005

• 论著 • 上一篇    下一篇

miR-224对hep3B细胞增殖和凋亡的影响

毛英,陈保华,李新建,姚斌   

  1. 鹰潭市解放军第一八四医院普外科,江西 鹰潭 335000
  • 出版日期:2015-07-30 发布日期:2015-12-09
  • 通信作者: 李新建 E-mail:764937718@qq.com

miR-224 affects Hep3B cell proliferation and apoptosis

MAO Ying, CHEN Baohua, LI Xinjian, YAO Bin   

  1. Department of General Surgery, the 184th Hospital of Chinese People's Liberation Army in Yingtan City, Yingtan Jiangxi 335000, China
  • Published:2015-07-30 Online:2015-12-09
  • Contact: LI Xinjian E-mail: 764937718@qq.com

摘要: 背景与目的:miR-224在肝细胞癌中呈过表达,并参与癌症的侵袭与转移过程,该研究通过观察miR-224体内、外对Hep3B细胞增殖和凋亡的影响,探讨miRNA-224与肝细胞癌发生、发展之间的关系。方法:运用实时荧光定量PCR定量分析miR-224反义寡核酸技术(antisense oligonucleotide,ASO)对Hep3B细胞mRNA表达的影响;通过miR-224 ASO降低Hep3B细胞中miR-224的表达,采用MTT、克隆形成实验、流式细胞术及体内实验观察miR-224 ASO对Hep3B细胞产生的生物学效应。结果:Hep3B细胞miR-224 ASO转染组与对照组相比,miR-224水平明显下降(P<0.05);MTT实验结果显示转染miR-224 ASO后,Hep3B细胞的存活数量明显低于对照组(P<0.05);克隆形成实验结果显示miR-224 ASO组克隆形成率较对照组显著降低(P<0.05);体内研究进一步证实miR-224 ASO可以抑制肿瘤细胞的增殖,从而导致肿瘤生长较对照组慢(P<0.05),肿瘤的体积较对照组明显减小(P<0.05);流式细胞术检测结果显示,转染miR-224 ASO组较对照组细胞凋亡明显增加(P<0.05);另外,降低miR-224的表达后发现Bcl-2的mRNA和蛋白都明显下降。结论:miR-224在Hep3B细胞中表达上调,降低miR-224的表达可有效抑制Hep3B细胞生长、促进细胞凋亡。miR-224有可能成为肝细胞癌表达调控的新靶点。

关键词:  miR-224, Hep3B, 反义单核苷酸, 细胞增殖, 细胞凋亡

Abstract: Background and purpose: MiR-224 is overexpressed in hepatocellular carcinoma, and participate in invasion and metastasis of cancer. The aim of this study was to investigate the effects of miR-224 antisense oligonucleotide (ASO) on the proliferation and apoptosis of Hep3B cells. Methods: After transfection with miR-224 ASO, and detecting the miR-224 mRNA expression of Hep3B cells by real-time quantitative PCR; the miR-224 expression in Hep3B cells was measured and cell proliferation was analyzed by MTT assay and the colony formation experiment in vitro and in vivo. The cell apoptosis was analyzed by flow cytometry. Results: Compared with the control group, miR-224 ASO significantly reduced the miR-224 mRNA expression in the Hep3B cell(P<0.05), MTT assay results showed that Hep3B cells survived rate decreased greatly after transfection with miR-224 ASO. Clone formation assay revealed that the colony formation rate in miR-224 ASO group was significantly lower than that in the control group. In vivo study further confirmed that miR-224 ASO could inhibit the proliferation of Hep3B cells, and miR-224 ASO group grew substantially slow compared with the negative control. Flow cytometry indicated that miR-224 ASO group promoted apoptosis significantly. Conclusion: miR- 224 was overexpressed in Hep3B cells. Reducing the expression of miR-224 can effectively inhibit the growth of Hep3B cells and promote apoptosis. miR-224 may become a new target for the regulation of gene expression in hepatocellular
carcinoma.

Key words: miR-224, Hep3B, Antisense oligonucleotides, Cell proliferation, Apoptosis