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LIM和SH3结构域蛋白1基因敲低通过抑制丝裂原活化蛋白激酶信号通路抑制结肠癌细胞的增殖和转移。

LIM and SH3 protein 1 knockdown suppresses proliferation and metastasis of colorectal carcinoma cells via inhibition of the mitogen-activated protein kinase signaling pathway.

作者信息

Zhao Hongpeng, Liu Bo, Li Jie

机构信息

Department of Gastrointestinal Surgery, Shandong Qianfoshan Hospital, Jinan, Shandong 250014, P.R. China.

Department of Hepatobiliary Surgery, Shandong Qianfoshan Hospital, Jinan, Shandong 250014, P.R. China.

出版信息

Oncol Lett. 2018 May;15(5):6839-6844. doi: 10.3892/ol.2018.8222. Epub 2018 Mar 9.

DOI:10.3892/ol.2018.8222
PMID:29731863
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC5920965/
Abstract

LIM and SH3 protein 1 (Lasp-1), a focal adhesion protein, serves a critical role in the regulation of cell proliferation and migration. The role of Lasp-1, as well as the intracellular signaling pathways involved in metastasis and progression of colorectal carcinoma, remains unclear. In the present study, the regulatory effect of Lasp-1 and the underlying molecular mechanism involved in migration and invasion of colorectal carcinoma were investigated. RNA interference and overexpression in SW480 cells were performed to elucidate the role of Lasp-1. Reverse transcription-quantitative polymerase chain reaction, western blotting and immunofluorescence were conducted to determine the mRNA and protein levels of Lasp-1 and extracellular-signal-regulated kinase 1/2 (ERK1/2) in SW480 cells as well as tumor and adjacent normal tissues obtained from 20 patients with colorectal carcinoma. Furthermore, a cell proliferation assay, flow cytometric analysis, and cell migration and invasion assays were performed to examine the effect of Lasp-1 on cell growth. The results of the present study demonstrated that Lasp-1 and ERK1/2 were upregulated in tumor tissue compared with adjacent normal colorectal mucosa. Cell-based assays demonstrated that Lasp-1 knockdown suppressed the expression and activation of ERK1/2, whereas Lasp-1 overexpression resulted in ERK1/2 upregulation. Additionally, Lasp-1 knockdown inhibited cell proliferation, migration, and invasion and induced cellular apoptosis and G/G cell-cycle arrest. The results of the present study indicate that Lasp-1 serves a critical role in the progression of colorectal carcinoma regulating the activation of the mitogen-activated protein kinase signaling pathway.

摘要

LIM和SH3结构域蛋白1(Lasp-1)是一种粘着斑蛋白,在细胞增殖和迁移的调控中发挥关键作用。Lasp-1的作用以及参与结直肠癌转移和进展的细胞内信号通路仍不清楚。在本研究中,研究了Lasp-1的调节作用以及参与结直肠癌迁移和侵袭的潜在分子机制。在SW480细胞中进行RNA干扰和过表达以阐明Lasp-1的作用。进行逆转录定量聚合酶链反应、蛋白质印迹和免疫荧光以确定SW480细胞以及从20例结直肠癌患者获得的肿瘤和相邻正常组织中Lasp-1和细胞外信号调节激酶1/2(ERK1/2)的mRNA和蛋白水平。此外,进行细胞增殖测定、流式细胞术分析以及细胞迁移和侵袭测定以检测Lasp-1对细胞生长的影响。本研究结果表明,与相邻正常结直肠黏膜相比,肿瘤组织中Lasp-1和ERK1/2上调。基于细胞的实验表明,Lasp-1敲低抑制ERK1/2的表达和激活,而Lasp-1过表达导致ERK1/2上调。此外,Lasp-1敲低抑制细胞增殖、迁移和侵袭,并诱导细胞凋亡和G/G期细胞周期阻滞。本研究结果表明,Lasp-1在结直肠癌进展中发挥关键作用,调节丝裂原活化蛋白激酶信号通路的激活。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4ef1/5920965/d2b497ac137d/ol-15-05-6839-g04.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4ef1/5920965/317673d926b4/ol-15-05-6839-g00.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4ef1/5920965/cc070de18db0/ol-15-05-6839-g01.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4ef1/5920965/6f318f4d25ff/ol-15-05-6839-g02.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4ef1/5920965/095fc1d81ac6/ol-15-05-6839-g03.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4ef1/5920965/d2b497ac137d/ol-15-05-6839-g04.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4ef1/5920965/317673d926b4/ol-15-05-6839-g00.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4ef1/5920965/cc070de18db0/ol-15-05-6839-g01.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4ef1/5920965/6f318f4d25ff/ol-15-05-6839-g02.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4ef1/5920965/095fc1d81ac6/ol-15-05-6839-g03.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4ef1/5920965/d2b497ac137d/ol-15-05-6839-g04.jpg

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