miR-182靶向作用环磷酸腺苷反应元件结合蛋白1调控人黑素瘤A375细胞的增殖和侵袭

王丽纳 王凯波 庄永灿 王红兰

解剖学报 ›› 2017, Vol. 48 ›› Issue (6) : 710-714.

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解剖学报 ›› 2017, Vol. 48 ›› Issue (6) : 710-714. DOI: 10.16098/j.issn.0529-1356.2017.06.013
肿瘤生物学

miR-182靶向作用环磷酸腺苷反应元件结合蛋白1调控人黑素瘤A375细胞的增殖和侵袭

  • 王丽纳1* 王凯波2 庄永灿1 王红兰1
作者信息 +

Effects of miR-182 on the proliferation and invasion of human melanoma cell line A375 by targeting of cyclic adenosine monophophate responsive element binding protein 1

  • WANG Li-na 1* WANG Kai-bo2 ZHUANG Yong-can1 WANG Hong-lan1
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文章历史 +

摘要

目的 探讨miR-182对人皮肤黑素瘤细胞系A375细胞的增殖与侵袭能力的影响,及miR-182影响黑色素细胞增殖的机制。 方法 利用阳离子脂质体LipofectamineTM2000将miR-182 模拟物或抑制剂转染人黑素瘤A375细胞,以使 miR-182过表达或低表达,采用MTT方法检测细胞活力变化,应用Transwell小室法检测A375细胞侵袭能力的变化,同时通过Real-time PCR检测核转录因子环磷酸腺苷反应元件结合蛋白1(CREB1) mRNA的表达情况,采用Western blotting检测CREB1蛋白的表达水平。 结果 与对照组相比,miR-182沉默能够显著增强A375细胞增殖和侵袭能力,P<0.01,显著降低CREB1蛋白与mRNA的表达水平,P<0.01;miR-182过表达的作用与之相反。 结论 miR-182沉默增强A375细胞增殖和侵袭能力可能与降低CREB1蛋白相关。

Abstract

Objective To investigate the effects of miR-182 on the proliferation and invasion of human melanoma cell line A375 and to elucidate its action mechanisms. Methods miR-182 mimics or inhibitor were transfected into A375 cells by lipofectamine package, MTT was applied to detect the viability of A375 cells, transwell chamber assay was used to measure the invasion of A375 cells, the expression levels of cyclic adenosine monophosphate responsive element binding protein 1(CREB1) mRNA and protein were detected by Real-time PCR and Western blotting respectively. Results Compared with the untreated group, miR-182 silence significantly increased the proliferation and invasion ability of A375 cells and obviously decreased the expression levels of CREB1 mRNA and protein, P<0.01, but with opposite role for miR-182 overexpression. Conclusion The reinforcing effect of miR-182 silence on the proliferation and invasion of A375 cells may be related to the down-regulation of CREB1.

关键词

miR-182 / 增殖 / 侵袭 / 环磷酸腺苷反应元件结合蛋白1 / 转染 / A375细胞 / 实时定量聚合酶链反应 /

Key words

miR-182 / Proliferation / Invasion / Cyclic adenosine monophosphate responsive element binding protein 1 / Transfect / A375 cell / Real-time PCR / Human

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王丽纳 王凯波 庄永灿 王红兰. miR-182靶向作用环磷酸腺苷反应元件结合蛋白1调控人黑素瘤A375细胞的增殖和侵袭[J]. 解剖学报. 2017, 48(6): 710-714 https://doi.org/10.16098/j.issn.0529-1356.2017.06.013
WANG Li-na WANG Kai-bo ZHUANG Yong-can WANG Hong-lan. Effects of miR-182 on the proliferation and invasion of human melanoma cell line A375 by targeting of cyclic adenosine monophophate responsive element binding protein 1[J]. Acta Anatomica Sinica. 2017, 48(6): 710-714 https://doi.org/10.16098/j.issn.0529-1356.2017.06.013

参考文献

[1]Mccourt C, Dolan O, Gormley G. Malignant melanoma: a pictorial review[J]. Ulster Med J,2014, 83(2): 103-110. 
[2]de Vries E, Houterman S, Janssen-Heijnen ML, et al. Up-to-date survival estimates and historical trends of cutaneous malignant melanoma in the south-east of The Netherlands[J]. Ann Oncol, 2007, 18(6): 1110-1116. 
[3]Hu N, Wang C, Clifford RJ, et al. Integrative genomics analysis of genes with biallelic loss and its relation to the expression of mRNA and micro-RNA in esophageal squamous cell carcinoma[J]. BMC Genomics, 2015, 16(l):732.
[4]Streicher KL, Zhu W, Lehmann KP, et al. A novel oncogenic role for the miRNA-506-514 cluster in initiating melanocyte transformation and promoting melanoma growth[J]. Oncogene, 2011, 31(12): 1558-1570.
[5]Mazar J, DeBlasio D, Govindarajan SS, et al. Epigenetic regulation of microRNA-375 and its role in melanoma development in humans[J]. FEBS Lett, 2011, 585(15): 2467-2476. [6]Nguyen T, Kuo C, Nicholl MB, et al. Downregulation of microRNA-29c is associated with hypermethylation of tumorrelated genes and disease outcome in cutaneous reelanoma[J]. Epigenetics, 20ll, 6(3): 388-394.
[7]Segura MF, Hanniford D, Menendez S, et al. Aberrant miR-182 expression promotes melanoma metastasis by repressing F0X03 and microphthalmia-associated transcription factor[J]. Proc Natl Acad Sci USA, 2009, 106(6):1814-1819.
[8]Zhang W, Qian P, Zhang X, et al. Autocrine/paracrine human growth hormone stimulated miRNA 96-182-183 cluster promotes epithelial-mesenchymal transition and invasion in breast cancer [J]. J Biol Chem, 2015, 290(22):13812-13829.
[9]Boniol M, Autier P, Boyle P, et al. Cutaneous melanoma attributable to sunbed use:systematic review and meta-analysis[J]. BMJ, 2012, 345:4757-4763.
[10]Li P, Sheng C, Huang L, et al. MiR-183/-96/-182 cluster is upregulated in most breast cancers and increases cell proliferation and migration[J]. Breast Cancer Res, 2014, 16(6): 473.
[11]Kong WQ, Bai R, Liu T, et al. MicroRNA-182 targets cAMPresponsive element-binding protein 1 and suppresses cell growth in human gastric adenocarcinoma[J]. FEBS J, 2012, 279(7):1252-1260.
[12]Wang S, Yang MH, Wang XY, et al. Increased expression of miRNA-182 in colorectal carcinoma: an independent and tissuespecific prognostic factor[J]. Int J Clin Exp Pathol, 2014, 7(6):3498-3503.
[13]Xu X, Wu J, Li S, et al. Downregulation of microRNA-182-5p contributes to renal cell carcinoma proliferation via activating the AKT/FOXO3a signaling pathway[J]. Mol Cancer, 2014, 13(3):109.
[14]Jin ZK, Li JJ, Du DL, et al. Down-regulation ofmiR-182 and miR-183 acts as potential predictor biomarkers in progression,metastasis,and poor prognosis of non-small cell lung cancer[J]. Int J Clin Exp Pathol,2016,9(3):3430-3435. 
[15]Wang TH,Yeh CT,Ho JY, et al. OncomiR miR-96 and miR-182 promote cell proliferation and invasion through targeting ephrinA5 in hepatocellular carcinoma[J]. Mol Carcinog,2016,55(4):366-375. 
[16 Benjamin S, Wu DY, Chen Z,et al.Integrative analysis identifies targetable CREB1/FoxA1 transcriptional co-regulation as a predictor of prostate cancer recurrence[J]. Nucleic Acids Res,2016,44(9):4105-4122.
[17]Pigazzi M, Ricotti E, German Q, et al. cAMP response element binding protein (CREB) overexpression CREB has been described as critical for leukemia progression[J]. Haematologica, 2007,92(10): 1435-1437.
[18]Stephen P, Moore G, Joshua K,et al.Transcription factors and DNA repair enzymes compete for damaged promoter sites[J]. J Biol Chem,2016, 291(11):5452-5460.

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