中国癌症杂志 ›› 2013, Vol. 23 ›› Issue (5): 341-346.doi: 10.3969/j.issn.1007-3969.2013.05.004

• 论著 • 上一篇    下一篇

miRNA-196b过表达对K562细胞增殖、凋亡及survivin、Cox-2表达的影响

尹虹,刘玥,郑文岭,宋艳斌,马文丽   

  1. 南方医科大学基因工程研究所,广东 广州 510515
  • 出版日期:2013-05-25 发布日期:2014-11-19
  • 通信作者: 马文丽 E-mail:339093503@qq.com
  • 基金资助:
    广东省科技计划项目(No: 2012B031800135);广东省自然基金项目(No: S2011020003140)

Effects of miRNA-196b overexpression on proliferation, apoptosis and survivin, Cox-2 expression of K562 cells

YIN Hong, LIU Yue, ZHENG Wen-ling, SONG Yan-bin, MA Wen-li   

  1. Institute of Genetic Engineering, Southern Medical University, Guangzhou Guangdong 510515, China
  • Published:2013-05-25 Online:2014-11-19
  • Contact: MA Wen-li E-mail: 339093503@qq.com

摘要:

背景与目的:BCR-ABL融合基因是慢性粒细胞白血病发病的分子病理基础,也是诊断慢性粒细胞白血病、观察疗效、评估预后等的有效指标。miRNA-196b在急性粒细胞白血病中低表达并对疾病的发展起主要作用。在慢性粒细胞白血病中miRNA-196b的靶基因为BCR-ABLmiRNA-196b过表达抑制BCR-ABL融合基因的表达。生存素(survivin)BCR-ABL的一个下游基因,已在多种肿瘤中发现survivin与环氧化酶-2(Cox-2)协同调节细胞的增殖凋亡。本研究旨在探讨miRNA-196b过表达对K562细胞增殖、凋亡及survivinCox-2 mRNA表达的影响。方法:实验分为K562-196b组、空载K562-pLV组与K562组,采用CCK-8法检测细胞增殖;采用Annexin-PE检测细胞凋亡;采用实时荧光定量PCR法检测Cox-2survivin mRNA的表达情况。结果:miRNA-196b过表达可以明显抑制K562细胞增殖;K562-196b组细胞凋亡率显著高于K562(P<0.05)miRNA-196b组中survivin基因显著低表达(P<0.05)Cox-2基因中无明显变化(P>0.05)结论:miRNA-196bK562细胞的增殖抑制和诱导凋亡有显著作用;miRNA-196b过表达可下调survivin基因的表达,为miRNA-196b作为慢性粒细胞性白血病的治疗靶点提供了依据。

关键词: miRNA-196b, K562细胞, 细胞增殖, 细胞凋亡

Abstract:

Background and purpose: BCR-ABL fusion gene is considered to be the molecular pathological basis and an effective indicator for diagnosis, observation, prognosis, and monitoring of chronic myelogenous leukemia (CML). MiRNA-196b had low expression in acute myeloid leukemia and played an important role in the development of disease. BCR-ABL is the target gene of miRNA-196b in CML, miRNA-196b overexpression leaded to BCR-ABL down-regulation or silencing. Survivin is a downstream gene of BCR-ABL signal pathways. Various studies had showed that survivin and Cox-2 cooperative regulated of cell proliferation and apoptosis in variety of tumors. The purpose of this study was to investigate the effects of miRNA-196b overexpression on proliferation, apoptosis and surviving, Cox-2 mRNA expression of K562 cells. Methods: Three groups including K562-196b, K562-pLV and K562 control groups were set up in this study. The cell proliferation and apoptosis were measured by CCK-8 assay and Annexin V-PE, respectively. The expression of Cox-2 and survivin genes at the mRNA level were detected by Q-PCR. Results: The proliferation of K562 cells could be significantly inhibited by miRNA-196b overexpression; Compared of the three groups of apoptosis rate, K562-196b group was significantly higher than K562 group (P<0.05). The expression of survivin gene in miRNA-196b was donwregulated (P<0.05), but the expression of Cox-2 gene in miRNA-196b group had no significant difference (P>0.05). Conclusion: The miRNA-196b plays an important role in K562 cells proliferation inhibition and apoptosis; Overexpression of miRNA-196b can down-regulate survivin gene expression, and provide some basis for miRNA-196b as a therapeutic target for chronic myelogenous leukemia.

Key words: MiRNA-196b, K562 cells, Cell proliferation, Cell apoptosis