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

doi:10.16098/j.issn.0529-1356.2017.06.013

解剖学报 ›› 2017, Vol. 48 ›› Issue (6): 710-714.doi: 10.16098/j.issn.0529-1356.2017.06.013

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

1. 福建医科大学附属泉州第一医院皮肤科，福建泉州 362000; 2. 沈阳市第七人民医院皮肤科，沈阳 110000

收稿日期:2017-03-03 修回日期:2017-05-10 出版日期:2017-12-06 发布日期:2017-12-06
通讯作者: 王丽纳 E-mail:naliwang008@163.com

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-bo²ZHUANG Yong-can¹ WANG Hong-lan¹

1. Department of Dermatology，the First Affiliated Hospital of Fujian Medical University in Quanzhou，Fujian Quanzhou 362000，China;2. Department of Dermatology，the Seventh People’s Hospital of Shenyang， Shenyang 110000，China

Received:2017-03-03 Revised:2017-05-10 Online:2017-12-06 Published:2017-12-06
Contact: WANG Li-na E-mail:naliwang008@163.com

摘要/Abstract

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Lipofectamine^TM2000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。

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, 人

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.

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

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.

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.

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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

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

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