miR-128-3p过表达靶向MAPK1抑制膀胱癌5637细胞株的侵袭，迁移和上皮间质转化

doi:10.13418/j.issn.1001-165x.2019.05.011

中国临床解剖学杂志 ›› 2019, Vol. 37 ›› Issue (5) : 534-541. DOI: 10.13418/j.issn.1001-165x.2019.05.011

赵阳，　万银绪，　车吉忠，　徐延凯，　李庆元

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Overexpression of miR-128-3p inhibits cell invasion, migration and epithelial-mesenchymal transition by targeting MAPK1 in bladder cancer cell line 5637

ZHAO Yang, WAN Yin-xu, CHE Ji-zhong, XU Yan-kai, LI Qing-yuan

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摘要

目的　研究miR-128-3p过表达对膀胱癌5637细胞株的侵袭，迁移和上皮间质转化影响。方法　基因预测软件TargetScan筛选出miR-128-3p的靶基因，荧光素酶报告实验验证；RT-PCR检测miR-128-3p和MAPK1的表达，Transwell检测细胞侵袭情况，划痕实验检测细胞迁移能力，Western blot检测E-cadherin、N-cadherin、ERK1/2、c-Myc和c-fos的表达，免疫荧光检测Vimentin的表达；裸鼠皮下注射建立移植瘤模型，30 d后检测瘤重量，绘制存活曲线，检测移植瘤中Vimentin、miR-128-3p、MAPK1、ERK1/2、c-Myc和c-fos的量。结果　miR-128-3p靶向抑制MAPK1表达；miR-128-3p过表达后，侵袭细胞数目、伤口愈合率降低，E-cadherin表达上调，N-cadherin表达下调，Vimentin阳性率减少，p-ERK1/2、c-Myc和c-fos表达下调。经miR-128-3p干预，裸鼠体内移植瘤重量减轻，存活率增加，miR-128-3p表达上调，MAPK1的表达下调，Vimentin阳性率减少，p-ERK1/2、c-Myc和c-fos表达下调。结论　过表达miR-128-3p通过靶向抑制MAPK1表达来抑制膀胱癌细胞5637的侵袭能力、迁移能力、上皮-间充质转化和ERK1/2、c-Myc和c-fos通路。

Abstract

Objective　To investigate the effects of miR-128-3p overexpression on invasion, migration and epithelial-mesenchymal transition of bladder cancer 5637 cells.　Methods　The target gene of miR-128-3p was screened by the gene prediction software TargetScan. And the luciferase reporter assay was verified. The expression of miR-128-3p and MAPK1 was detected by RT-PCR. Cell invasion was detected by Transwell. Cell migration ability was measured by a scratch test. The expressions of E-cadherin, N-cadherin, ERK1/2, c-Myc and c-fos were detected by Western blot, and the expression of Vimentin was detected by immunofluorescence. The xenograft model was established by subcutaneous injection in nude mice. The tumor weight was measured 30 days later. And the survival curve was drawn. The amount of Vimentin, miR-128-3p, MAPK1, ERK1/2, c-Myc and c-fos in the transplanted tumor was detected.　Results　miR-128-3p had been targeted to inhibit the expression of MAPK1. After overexpression of miR-128-3p, the number of invasive cells and wound healing rate was decreased. E-cadherin expression was up-regulated. N-cadherin expression was down-regulated. Vimentin positive was decreased. The expression of ERK1/2, c-Myc and c-fos was down-regulated. Through the intervention of miR-128-3p, the weight of transplanted tumors was decreased. The survival rate of nude mice was increased. The expression of miR-128-3p was up-regulated. The expression of MAPK1 was down-regulated. The expression of Vimentin was decreased. And the expression of ERK1/2, c-Myc and c-fos was decreased.　Conclusion　Overexpression of miR-128-3p inhibits the invasion, migration, epithelial-mesenchymal transition and ERK1/2, c-Myc and c-fos pathways of bladder cancer cell line 5637 by targeting inhibition of MAPK1 expression.

导出引用

赵阳, 万银绪, 车吉忠, 徐延凯, 李庆元. miR-128-3p过表达靶向MAPK1抑制膀胱癌5637细胞株的侵袭，迁移和上皮间质转化[J]. 中国临床解剖学杂志. 2019, 37(5): 534-541 https://doi.org/10.13418/j.issn.1001-165x.2019.05.011

ZHAO Yang, WAN Yin-xu, CHE Ji-zhong, XU Yan-kai, LI Qing-yuan. Overexpression of miR-128-3p inhibits cell invasion, migration and epithelial-mesenchymal transition by targeting MAPK1 in bladder cancer cell line 5637[J]. Chinese Journal of Clinical Anatomy. 2019, 37(5): 534-541 https://doi.org/10.13418/j.issn.1001-165x.2019.05.011

中图分类号： R734.2

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