上调的miR-452-5p抑制RORα的表达并促进肝癌细胞的增殖和迁移
Up-regulated miR-452-5p suppressed the expression of RORα and promoted the proliferation and migration of HCC cells
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摘要: 为了探究肝细胞癌中miR-452-5p对患者预后生存的影响及其对肝癌细胞增殖、迁移的作用。采用肿瘤基因组图谱（TCGA）数据库中肝细胞肝癌数据集对miR-452-5p进行差异表达分析和Kaplan-Meier生存分析。采用TargetscanHuman和miRDB靶基因数据库对miR-452-5p靶基因进行预测；采用基因差异表达分析和加权基因共表达网络分析（WGCNA）的方法对数据集GSE14520进行分析计算。用Lipofectmine-2000将miR-452-5p模拟物、模拟物阴性对照和miR-452-5p抑制剂、抑制剂阴性对照分别转染到Huh7细胞中。分别采用RT-qPCR和Western blot实验，检测4组细胞中RORα在mRNA和蛋白水平上的表达情况。采用CCK-8、Transwell实验，检测4组细胞的增殖活力以及迁移能力。双荧光素酶报告基因实验验证Huh7细胞中miR-452-5p与RORα的调控关系。经TCGA数据分析，miR-452-5p在肝癌组织中高表达且显著影响患者预后总生存期。通过miRNA靶基因预测、基因差异表达分析、WGCNA分析，筛选出关键基因RORα和LAMC1。通过Kaplan-Meier生存分析得知RORα的异常表达显著影响肝癌患者的预后总生存期。肝癌细胞中miR-452-5p的过表达，会降低RORα的mRNA和蛋白表达，同时增强肝癌细胞的增殖能力和迁移能力。双荧光素酶报告基因实验结果证实了miR-452-5p靶向RORα的3′UTR区。在肝细胞癌患者中高表达的miR-452-5p靶向抑制了RORα的表达，促进了肝癌细胞增殖和迁移，进而加剧了肝癌的进展和预后不良。Abstract: To study the prognosis-related regulation mechanism of miR-452-5p and its influence on the proliferation and migration of hepatocellular carcinoma (HCC) cells, liver hepatocellular carcinoma (LIHC) dataset in The Cancer Genome Atlas (TCGA) was used to validate the differential expression of miR-452-5p and perform the Kaplan-Meier analysis of overall survival (OS).Target genes of miR-452-5p from TargetscanHuman and miRDB databases were predictived; and differentially expressed genes(DEGs) and weighted gene co-expression network analysis (WCGNA) were completed with GSE14520.Lipofectmine-2000 was used to transfect miR-452-5p mimics, mimics negative control, miR-452-5p inhibitor and inhibitor negative control into Huh7 cells,respectively.The mRNA and protein expression level of RORα in 4 groups were determined by RT-qPCR and Western blot.CCK-8 assay and Transwell assay were conducted to testify the capabilities of proliferation and migration.The regulation between miR-452-5p and RORα was confirmed by the dual luciferase reporter assay.After analysis in the TCGA-LIHC dataset, miR-452-5p had higher expression in HCC tissue than that in normal tissue, which was also associated with a shorter OS.RORα and LAMC1 were discriminated by intersecting of DEGs, WGCNA module genes, and predictive target genes.Survival analysis exhibited that dysregulation of RORα was significantly related to the OS.Overexpression of miR-452-5p in HCC cells suppressed the expression of RORα both in mRNA and protein, and also enhanced the viability and migration of HCC cells.The results of the dual luciferase reporter assay showed that miR-452-5p targeted 3′UTR of RORα.Up-regulated miR-452-5p inhibited the expression of RORα, facilitated the proliferation and migration of HCC cells, and promoted the progression and poor prognosis of HCC.
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Keywords:
- miR-452-5p /
- hepatocellular carcinoma /
- bioinformatics /
- differential expression /
- overall survival /
- RORα /
- proliferation /
- migration
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[1] . CA Cancer J Clin，2018，68（6）：394-424. [2] El-Serag HB，Rudolph KL. Hepatocellular carcinoma：epidemiology and molecular carcinogenesis［J］. Gastroenterology，2007，132（7）：2557-2576. [3] Hsu YC，Yip TC，Ho HJ，et al. Development of a scoring system to predict hepatocellular carcinoma in Asians on antivirals for chronic hepatitis B［J］. J Hepatol，2018，69（2）：278-285. [4] Chang KC，Wu YY，Hung CH，et al. Clinical-guide risk prediction of hepatocellular carcinoma development in chronic hepatitis C patients after interferon-based therapy［J］. Br J Cancer，2013，109（9）：2481-2488. [5] Waziry R，Hajarizadeh B，Grebely J，et al. Hepatocellular carcinoma risk following direct-acting antiviral HCV therapy：a systematic review，meta-analyses，and meta-regression［J］. J Hepatol，2017，67（6）：1204-1212. [6] Kulik L，El-Serag HB. Epidemiology and management of hepatocellular carcinoma［J］. Gastroenterology，2019，156（2）：477-491. el. [7] McGlynn KA，Petrick JL，El-Serag HB. Epidemiology of hepatocellular carcinoma［J］. Hepatology，2021，73（ Suppl 1 ）：4-13.[8] Pi?ero F，Dirchwolf M，Pess?a MG. Biomarkers in hepatocellular carcinoma：diagnosis，prognosis and treatment response assessment［J］. Cells，2020，9（6）： E1370 .[9] Bartel DP. microRNAs：target recognition and regulatory functions［J］. Cell，2009，136（2）：215-233. [10] Lu TX，Rothenberg ME. microRNA［J］. J Allergy Clin Immunol，2018，141（4）：1202-1207. [11] Krol J，Loedige I，Filipowicz W. The widespread regulation of microRNA biogenesis，function and decay［J］. Nat Rev Genet，2010，11（9）：597-610. [12] Ali SZ，Langden SSS，Munkhzul C，et al. Regulatory mechanism of microRNA expression in cancer［J］. Int J Mol Sci，2020，21（5）：1723. [13] Lin X，Han L，Gu CC，et al. miR-452-5p promotes colorectal cancer progression by regulating an ERK/MAPK positive feedback loop［J］. Aging （Albany NY），2021，13（5）：7608-7626. [14] Zhai W，Li SY，Zhang J，et al. Sunitinib-suppressed miR-452-5p facilitates renal cancer cell invasion and metastasis through modulating SMAD4/SMAD7 signals［J］. Mol Cancer，2018，17（1）：157. [15] Zheng QL，Sheng Q，Jiang CY，et al. microRNA-452 promotes tumorigenesis in hepatocellular carcinoma by targeting cyclin-dependent kinase inhibitor 1B［J］. Mol Cell Biochem，2014，389（1/2）：187-195. [16] Agarwal V，Bell GW，Nam JW，et al. Predicting effective microRNA target sites in mammalian mRNAs［J］. Elife，2015，4： e05005 .[17] Chen YH，Wang XW. miRDB：an online database for prediction of functional microRNA targets［J］. Nucleic Acids Res，2020，48（D1）：D127-D131. [18] Langfelder P，Horvath S. WGCNA：an R package for weighted correlation network analysis［J］. BMC Bioinformatics，2008，9：559. [19] Yi M，Xu LP，Jiao Y，et al. The role of cancer-derived microRNAs in cancer immune escape［J］. J Hematol Oncol，2020，13（1）：25. [20] Rupaimoole R，Slack FJ. microRNA therapeutics：towards a new era for the management of cancer and other diseases［J］. Nat Rev Drug Discov，2017，16（3）：203-222. [21] Gao L，Zhang LJ，Li SH，et al. Role of miR-452-5p in the tumorigenesis of prostate cancer：a study based on the Cancer Genome Atl（TCGA），Gene Expression Omnibus （GEO），and bioinformatics analysis［J］. Pathol Res Pract，2018，214（5）：732-749. [22] Zhang Y，Li X，Zhang J，et al. Circ-CCDC66 upregulates REXO1 expression to aggravate cervical cancer progression via restraining miR-452-5p［J］. Cancer Cell Int，2021，21（1）：20. [23] Mao W，Xiong GF，Wu YY，et al. RORα suppresses cancer-associated inflammation by repressing respiratory complex I-dependent ROS generation［J］. Int J Mol Sci，2021，22（19）：10665. [24] Liu G，Yang ZF，Zhou PY，et al. ROR-α-1 inhibits the proliferation，invasion，and migration of hepatocellular carcinoma MHCC97H via downregulation of chemokine CXCL5［J］. Cytokine，2020，129：155004. [25] Huang JL，Fu YP，Gan W，et al. Hepatic stellate cells promote the progression of hepatocellular carcinoma through microRNA-1246-RORα-Wnt/β-Catenin axis［J］. Cancer Lett，2020，476：140-151. [26] Han LL，Huang C，Zhang SQ. The RNA-binding protein SORBS2 suppresses hepatocellular carcinoma tumourigenesis and metastasis by stabilizing RORA mRNA［J］. Liver Int，2019，39（11）：2190-2203. -
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