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MicroRNA-10a通过靶向作用E2F3抑制肝癌细胞的增殖 被引量:1

MicroRNA-10a inhibits hepatocellular carcinoma cell proliferation through targeting E2F3
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摘要 目的:探讨微小RNA-10a(microRNA-10a,miR-10a)对肝癌细胞增殖的影响及其作用机制。方法:收集广西医科大学附属肿瘤医院肿瘤科2001年10月至2005年7月144例肝癌患者手术切除的肝癌组织和癌旁组织(距癌灶组织边缘2~5 cm)标本,Real-time PCR法分析144例肝癌组织及癌旁组织中miR-10a的表达量。在肝癌细胞(QGY-7701、Huh7、PCL/PRF/5)中转染miR-10a模拟物,Real-time PCR法检测转染后细胞miR-10a的表达水平;CCK-8法检测过表达miR-10a的肝癌细胞的增殖水平,流式细胞术检测过表达miR-10a的肝癌细胞的凋亡和细胞周期;生物信息学预测并以Western blotting检测过表达miR-10a的肝癌细胞中转录因子E2F3的表达量。结果:与癌旁组织相比,肝癌组织中的miR-10a显著低表达[(-9.89±1.68)vs(-7.84±1.97),P=0.000]。转染miR-10a模拟物后肝癌细胞系中miR-10a的表达量是转染对照小RNA组或空白组细胞的16倍左右。过表达miR-10a可显著抑制7种肝癌细胞(QGY-7701、QGY-7703、Huh7、PCL/PRF/5、HepG2、BeL-7402、SMMC-7721)的增殖(均P〈0.05),并引起肝癌细胞细胞周期G1/S期阻滞,但并不能诱导肝癌细胞发生凋亡。生物信息学预测显示E2F3是miR-10a可能的靶分子,Western blotting检测显示过表达miR-10a可明显抑制肝癌细胞中E2F3的表达[(0.50±0.12)vs(0.79±0.21),P〈0.05]。结论:人肝癌组织中低表达miR-10a,转染miR-10a模拟物后多种肝癌细胞的增殖均受到明显抑制,其机制可能与miR-10a靶向作用转录因子E2F3并阻滞肝癌细胞细胞周期于G1/S期有关。 Objective: To investigate the role of microRNA-10a( miR-10a) in hepatocellular carcinoma( HCC)growth. Methods: Paired HCC and adjacent non-tumor tissue specimens were surgically collected from 144 patients who were diagnosed with primary HCC in Guangxi Medical University-Affiliated Tumor Hospital between October 2001 and July2005. HCC QGY-7701,Huh7,and PCL /PRF /5 cells were transfected with miR-10 a mimics or scramble control miRNA.The abundance of miR-10 a in both tissue specimens and transfected cells was quantified by real-time PCR and E2F3 protein in transfected cells was assessed by Western blotting. Proliferation of the transfectants was assessed by a colorimetric cell counting assay. Cell cycle progression and apoptosis of the transfectants were assessed by FACS. Results: The abundance of miR-10 a mRNA was significantly lower in HCC tissue specimens than in normal tissue specimens(- 9. 89 ±1. 68 vs- 7. 84 ± 1. 97,P = 0. 0001). HCC cells transfected with miR-10 a mimics had miR-10 a abundance 16 times higher than both wild-type HCC cells and HCC cells transfected with the control miRNA with scrambled sequences. Over-expression of miR-10 a resulted in significant increases in suppression of HCC cell proliferation( P 0. 05) and G1 phase arrest. In contrast,overexpression of miR-10 a had no influence on apoptosis of HCC cells. Bioinformatics suggested that transcription factor E2F3 might be a downstream target of miR-10 a and the expression of E2F3 in HCC cells transfected with miR-10 a was significantly lower than in wild-type HCC cells and HCC cells transfected with the control miRNA( 0. 50 ±0.12 vs 0.79 ±0.21,P 0.05). Conclusion: MiR-10 a may suppress HCC cell proliferation through G1 phase arrest in an E2F3-dependent mechanism.
作者 赵锴 王春梅 曹雪涛 Zhao Kai, Wang Chunmei, Cao Xuetao (1. Institute of Basic Medical Sciences, Chinese Academy of Medical Sci- ences, Peking Union Medical College, Beijing 100006, China; 2. National Key Laboratory of Medical Immunology & Institute of Immunology, Second Military Medical University, Shanghai 200433, China)
出处 《中国肿瘤生物治疗杂志》 CAS CSCD 北大核心 2014年第4期383-388,共6页 Chinese Journal of Cancer Biotherapy
基金 国家高技术研究发展计划(863计划)资助项目
关键词 肝癌 微小RNA-10a 增殖 转录因子 E2F3 G1 S期阻滞 hepatocellular carcinoma microRNA-10a(miR-10a) transcription factor E2F3 proliferation G1/S phase arrest
作者简介 赵锴(1988-),男,陕西省汉中市人,硕士生,主要从事表观遗传学在肿瘤免疫中调控作用的研究,E-mail:zhaokail6@163.com 王春梅(Wang Chunmei,co-corresponding author),E—mail:wangcm1977@126.com。共同通信作者 曹雪涛(Cao Xuetao,corresponding author),E-mail:caoxt@immunol.org;
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