microRNA-1304通过靶向血红素氧合酶-1对人肺癌细胞的抑制作用

doi:10.12092/j.issn.1009-2501.2019.04.004

中国临床药理学与治疗学 ›› 2019, Vol. 24 ›› Issue (4): 383-390.doi: 10.12092/j.issn.1009-2501.2019.04.004

microRNA-1304通过靶向血红素氧合酶-1对人肺癌细胞的抑制作用

朱林佳¹，原少斐¹，上官宗校¹，慈晓¹，赵仁国¹，李玉苹²

¹温州医科大学附属第三医院呼吸科，²温州医科大学附属第一医院呼吸科，温州 325200，浙江

收稿日期:2018-08-07 修回日期:2018-10-13 出版日期:2019-04-26 发布日期:2019-05-01
通讯作者: 李玉苹，女，硕士，主任医师，研究方向：肺癌靶向治疗。 Tel:13868467423 E-mail:ysf1004@163.com
作者简介:朱林佳，女，硕士，主治医师，研究方向：肺癌靶向治疗。 Tel:13906871971 E-mail:kitty.linjia@163.com
基金资助:
浙江省医药卫生科技青年人才计划项目（2016101300）

miR-1304 inhibits non-small cell lung cancer cell invasion and proliferation in vitro via targeting heme oxygenase-1

ZHU Linjia¹, YUAN Shaofei ¹, SHANGGUAN Zongxiao¹,CI Xiao ¹, ZHAO Renguo ¹, LI Yuping ²

¹Department of Respiratory Medcine, the Third Affiliated Hospital, Wenzhou Medical University, Wenzhou 325200, Zhejiang, China；² Department of Respiratory Medcine, the First Affiliated Hospital, Wenzhou Medical University, Wenzhou 325200, Zhejiang, China

Received:2018-08-07 Revised:2018-10-13 Online:2019-04-26 Published:2019-05-01

摘要/Abstract

摘要：

目的:探讨miR-1304对肺癌细胞增殖和侵袭的作用及机制。方法:应用实时定量PCR检测miR-1304在不同转移能力肺癌A549和NCI-H1975细胞中的表达水平；脂质体2000介导分别转染miR-1304类似物和抑制物入A549 和 NCI-H1975细胞；应用MTT法和细胞集落形成实验分别检测miR-1304对肺癌细胞增殖活性的作用及生长抑制作用；细胞凋亡和细胞周期分别采用膜联蛋白V-PE/7-AAD和PI染色分析进行检测。双荧光素酶报告基因验证miR-1304是否作用于血红素氧合酶（heme oxygenase-1，HO-1） mRNA的3'UTR区预测靶位；Western blot检测HO-1蛋白水平。结果:miR-1304高表达可显著抑制肺癌细胞形成的数量和生存能力，以及诱导细胞凋亡和G0/G1期细胞周期阻滞。体外侵袭实验及细胞增殖实验结果显示，miR-1304可抑制肺癌细胞侵袭和增殖；经双荧光素酶报告基因验证HO-1是miR-1304的靶基因。结论:miR-1304在肺癌中表达，通过直接作用于HO-1可抑制肺癌细胞的侵袭和增殖，miR-1304是肺癌的一个潜在治疗靶点。

关键词: 肺癌, microRNA-1304, 血红素氧合酶-1, 侵袭, 增殖

Abstract:

AIM:To investigate the direct target of miR-1304 and its function in NSCLC in vitro. METHODS: Real-time quantitative PCR was used to detect the expression of miR-1304 in lung cancer cell A549 and NCI-H1975 .Transwell assay was used to test the role of miR-1304 on regulating invasion and migration of cells. The cell proliferation and survival were investigated via cell counting, MTT and colony-formation assays. Cell apoptosis and cell cycle were examined using annexin V-PE/7-AAD and PI staining assays, respectively. Dual luciferase reporter gene was used to analyze the binding between miR-1304 and 3'UTR of heme oxygenase-1, Western blot detected the HO-1 protein levels. RESULTS:MiR-1304 significantly decreased the number and viability of NSCLC cells and colony formation, and induced cell apoptosis and G0/G1 phase cell cycle arrest. HO-1 was demonstrated to be a direct target of miR-1304 in NSCLC cells. Furthermore, altered expression of miR-1304 by transfection of pre-miR-1304 mimics and inhibitor signifcantly affected the ability of invasion and proliferation of lung cancer cells. Altered expression of miR-1304 markedly down-regulated the HO-1 protein levels of lung cancer cells. In addition, dual luciferase reporter gene assay indicated that miR-1304 regulated HO-1 expression by binding to the 3'UTR of HO-1 mRNA. CONCLUSION: MicroRNA-1304 is a tumor suppressor and HO-1 is its direct target in NSCLC. The results suggest the potential for miR-1304 as a therapeutic target for NSCLC.

Key words: NSCLC, miR-1304, heme oxygenase-1, invasion, proliferation

中图分类号:

R965.2

朱林佳，原少斐，上官宗校，慈晓，赵仁国，李玉苹. microRNA-1304通过靶向血红素氧合酶-1对人肺癌细胞的抑制作用[J]. 中国临床药理学与治疗学, 2019, 24(4): 383-390.

Yuan ShaoFei. miR-1304 inhibits non-small cell lung cancer cell invasion and proliferation in vitro via targeting heme oxygenase-1[J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2019, 24(4): 383-390.

参考文献

［1］Wood SL, Pernemalm M, Crosbie PA, et al. The role of the tumor-micro environment in lung cancer-metastasis and its relationship to potential therapeutic targets［J］. Cancer Treat Rev, 2014, 40(8):558-566.
［2］Liu K, Li X, Cao Y, et al. miR-132 inhibits cell proliferation, invasion and migration of hepatocellular carcinoma by targeting PIK3R3［J］. Int J Oncol, 2015,47(9): 1585-1593.
［3］Liu N, Olson EN. MicroRNA regulatory networks in cardiovascular development［J］. Dev Cell, 2016, 18(5):510-525.
［4］Fabian MR, Sonenberg N, Filipowicz W. Regulation of mRNA translation and stability by microRNAs［J］. Annu Rev Biochem, 2010, 79(3):351-379.
［5］Bartel DP. MicroRNAs: target recognition and regulatory functions［J］.Cell,2015,136(6): 215-233.
［6］Fabbri M. miRNAs as molecular biomarkers of cancer［J］. Expert Rev Mol Diagn,2010, 10(4):435-444.
［7］Lopez-Valenzuela M, Ramirez O, Rosas A, Garcia-Vargas S, et al. An ancestral miR-1304 allele present in Neanderthals regulates genes involved in enamel formation and could explain dental differences with modern humans［J］. Mol Biol Evol, 2012,29(11):1797-1806.
［8］Jemal A，Bray F，Center MM，et al．Global cancer statistics［J］． CA Cancer J Clin，2011，61(2):69-90.
［9］Barad O，Meiri E，Avniel A，et al．MicroRNA expression detected by oligonucleotide microarrays: system establishment and expression profiling in human tissues［J］．Genome Res，2014，14(12) :2486-2494.
［10］徐明,崔鹏,叶敏,等.miRNA-101在甲状腺髓样癌中抑癌机制的研究［J］.中国临床药理学与治疗学,2018,23（5）:524-530.
［11］Lee I，Ajay SS，Yook JI，et al．New class of microRNA targets containing simultaneous 5'-UTR and 3'-UTR interaction sites［J］．Genome Res，2016，19(7):1175-1183．
［12］Calin GA，Sevignani C，Dumitru CD，et al．Human microRNA genes are frequently located at fragile sites and genomic regions involved in cancers［J］．Proc Natl Acad Sci USA，2004,101(9):2999-3004.
［13］梅颖, 刘朝奇.miRNA调控肿瘤相关基因c-met的研究进展［J］.中国临床药理学与治疗学,2017,22（2）:204-209.
［14］Vaz C, Ahmad HM, Sharma P, et al. Analysis of microRNA transcriptome by deep sequencing of small RNA libraries of peripheral blood［J］. BMC Genomics, 2010, 11(6):288-293.
［15］Stark MS, Tyagi S, Nancarrow DJ,et al.Characterization of the melanoma miRNAome by deep sequencing［J］. PLoS One, 2010, 5: e9685.
［16］Xu CZ, Shi RJ, Chen D, et al. Potential biomarkers for paclitaxel sensitivity in hypopharynx cancer cell［J］. Int JClin Exp Pathol,2013, 6(12): 2745-2756.
［17］Hsu FF, Yeh CT, Sun YJ, et al. Signal peptide peptidase-mediated nuclear localization of heme oxygenase-1 promotes cancer cell proliferation and invasion independent of its enzymatic activity［J］. Oncogene, 2015,34(11): 2360-2370.
［18］Pittala V, Salerno L, Romeo G, et al. A focus on heme oxygenase-1 (HO-1) inhibitors［J］. Curr Med Chem, 2013, 20(10):3711-3732.
［19］Gao C, Peng FH, Peng LK. MiR-200c sensitizes clear-cell renal cell carcinoma cells to sorafenib and imatinib by targeting heme oxygenase-1［J］. Neoplasma, 2014,61(9): 680-689.
［20］Xiao S, Wang X, Ni H, et al. MicroRNA miR-24-3p promotes porcine reproductive and respiratory syndrome virus replication through suppression of heme oxygenase-1 expression［J］. J Virol, 2015,89(10): 4494-4503.

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microRNA-1304通过靶向血红素氧合酶-1对人肺癌细胞的抑制作用

miR-1304 inhibits non-small cell lung cancer cell invasion and proliferation in vitro via targeting heme oxygenase-1

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