MiR-142-5p通过靶向IGF2BP3调控多柔比星诱导的肝癌细胞凋亡

发布时间：2018-04-25 11:23

本文选题：肝细胞癌 + miR--p　；参考：《中国肿瘤生物治疗杂志》2017年06期

【摘要】：目的:探讨miR-142-5p对多柔比星诱导的原发性肝细胞癌(hepatocellular carcinoma,HCC)细胞凋亡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88例HCC患者手术切除的癌组织及癌旁组织(距癌灶组织边缘2~5 cm)标本。采用实时荧光定量PCR检测人HCC组织和癌旁组织、人正常肝细胞以及HCC细胞系中miR-142-5p的表达量。向HCC细胞SMMC-7721中转染miR-142-5p mimics,流式细胞术检测过表达miR-142-5p后SMMC-7721细胞在多柔比星(doxorubicin)(1μg/ml)诱导下凋亡的变化;生物信息学方法预测miR-142-5p可靶向结合胰岛素样生长因子2 mRNA结合蛋白3(insulin-like growth factor 2 mRNA-binding protein 3,IGF2BP3)基因,并采用荧光素酶报告基因实验进行验证。采用实时荧光定量PCR及Western blotting检测过表达miR-142-5p的SMMC-7721细胞中IGF2BP3的mRNA及蛋白表达情况。结果:与癌旁组织和正常肝细胞相比,HCC组织(-6.91±2.61 vs-11.59±2.59,P0.01)和多种HCC细胞系中miR-142-5p呈明显低表达(均P0.01);过表达miR-142-5p可显著促进多柔比星诱导的HCC细胞SMMC-7721的凋亡[(49.40±3.47)%vs(19.50±1.74)%,P0.01];过表达miR-142-5p可明显降低HCC细胞中IGF2BP3的mRNA及蛋白表达水平(P0.01),敲减IGF2BP3表达可进一步促进多柔比星诱导的SMMC-7721细胞的凋亡(P0.01)。荧光素酶报告基因实验结果显示,miR-142-5p能够抑制IGF2BP3的3'UTR荧光素酶报告基因的活性。结论:miR-142-5p在HCC组织标本和体外培养细胞系中的表达水平均显著降低,转染miR-142-5p mimics后能够促进多柔比星诱导的HCC细胞的凋亡,其机制可能与miR-142-5p靶向作用IGF2BP3从而促进HCC细胞凋亡有关。
[Abstract]:Aim: to investigate the effect of miR-142-5p on apoptosis of primary hepatocellular carcinoma (HCC) induced by doxorubicin and its mechanism. Methods: the specimens of cancer tissues and adjacent tissues (2cm to the margin of cancer tissue) were collected from 88 patients with HCC in affiliated Cancer Hospital of Guangxi Medical University. The expression of miR-142-5p in human HCC tissues, adjacent tissues, human normal hepatocytes and HCC cell lines was detected by real-time fluorescence quantitative PCR. MiR-142-5p mimics were transfected into SMMC-7721 of HCC cells. The apoptosis of SMMC-7721 cells induced by doxorubicinine 1 渭 g / ml was detected by flow cytometry. MiR-142-5p targeting insulin-like growth factor 2 mRNA binding protein 3(insulin-like growth factor 2 mRNA-binding protein 3 IGF2BP3) gene was predicted by bioinformatics and verified by luciferase reporter gene experiment. Real time quantitative PCR and Western blotting were used to detect the mRNA and protein expression of IGF2BP3 in SMMC-7721 cells. Results: compared with paracancerous tissues and normal hepatocytes, the expression of miR-142-5p was significantly lower in HCC tissues (-6.91 卤2.61 vs-11.59 卤2.59 vs-11.59 卤2.59 P0.01) and in various HCC cell lines (all P0.01C). Overexpression of miR-142-5p could significantly promote SMMC-7721 apoptosis induced by doxorubicin [49.40 卤3.47)%vs(19.50 卤1.74P0.01], and overexpression of miR-142-5p could significantly promote the apoptosis of SMMC-7721 in HCC cells induced by doxorubicin [49.40 卤3.47)%vs(19.50 卤1.74P0.01]. Reducing the expression of IGF2BP3 mRNA and protein in HCC cells could promote the apoptosis of SMMC-7721 cells induced by doxorubicin. The results of luciferase reporter gene experiment showed that miR-142-5p could inhibit the activity of 3'UTR luciferase reporter gene of IGF2BP3. Conclusion the expression level of 1: miR-142-5p in HCC tissue samples and in vitro cultured cell lines was significantly decreased. Transfection of miR-142-5p mimics could promote the apoptosis of HCC cells induced by doxorubicin. The mechanism may be related to the targeting of miR-142-5p to IGF2BP3 and thus to the apoptosis of HCC cells.
【作者单位】：中国医学科学院北京协和医学院基础医学研究所;第二军医大学免疫学研究所暨医学免疫学国家重点实验室;广西医科大学附属肿瘤医院;
【基金】：国家自然科学基金资助项目(No.31390431) 国家重点基础研究发展规划(973计划)资助项目(No.2013CB530503)~~
【分类号】：R735.7

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