Effects of metastasis-associated in colon cancer 1 inhibition by small hairpin RNA on ovarian carcinoma OVCAR-3 cells

<p>Abstract</p> <p>Background</p> <p>Metastasis-associated in colon cancer 1 (MACC1) is demonstrated to be up-regulated in several types of cancer, and can serve as biomarker for cancer invasion and metastasis. To investigate the relations between MACC1 and biological p...

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Main Authors: Ren Fang, Wu Qinghua, Chen Zhimin, Shi Huirong, Zhang Ruitao, Huang Haoliang
Format: Article
Language:English
Published: BMC 2011-09-01
Series:Journal of Experimental & Clinical Cancer Research
Subjects:
Online Access:http://www.jeccr.com/content/30/1/83
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author Ren Fang
Wu Qinghua
Chen Zhimin
Shi Huirong
Zhang Ruitao
Huang Haoliang
author_facet Ren Fang
Wu Qinghua
Chen Zhimin
Shi Huirong
Zhang Ruitao
Huang Haoliang
author_sort Ren Fang
collection DOAJ
description <p>Abstract</p> <p>Background</p> <p>Metastasis-associated in colon cancer 1 (MACC1) is demonstrated to be up-regulated in several types of cancer, and can serve as biomarker for cancer invasion and metastasis. To investigate the relations between MACC1 and biological processes of ovarian cancer, MACC1 specific small hairpin RNA (shRNA) expression plasmids were used to investigate the effects of MACC1 inhibition on ovarian carcinoma OVCAR-3 cells.</p> <p>Methods</p> <p>Expressions of MACC1 were detected in different ovarian tissues by immunohistochemistry. MACC1 specific shRNA expression plasmids were constructed and transfected into OVCAR-3 cells. Then, expressions of MACC1 were examined by reverse transcription polymerase chain reaction (RT-PCR) and Western blot. Cell proliferation was observed by MTT and monoplast colony formation assay. Flow cytometry and TUNEL assay were used to measure cell apoptosis. Cell migration was assessed by wound healing and transwell migration assay. Matrigel invasion and xenograft model assay were performed to analyze the potential of cell invasion. Activities of Met, MEK1/2, ERK1/2, Akt, cyclinD1, caspase3 and MMP2 protein were measured by Western blot.</p> <p>Results</p> <p>Overexpressions of MACC1 were detected in ovarian cancer tissues. Expression of MACC1 in OVCAR-3 cells was significantly down-regulated by MACC1 specific small hairpin RNA. In OVCAR-3 cells, down-regulation of MACC1 resulted in significant inhibition of cell proliferation, migration and invasion, meanwhile obvious enhancement of apoptosis. As a consequence of MACC1 knockdown, expressions of Met, p-MEK1/2, p-ERK1/2, cyclinD1 and MMP2 protein decreased, level of cleaved capase3 was increased.</p> <p>Conclusions</p> <p>RNA interference (RNAi) against MACC1 could serve as a promising intervention strategy for gene therapy of ovarian carcinoma, and the antitumor effects of MACC1 knockdown might involve in the inhibition of HGF/Met and MEK/ERK pathways.</p>
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spelling doaj.art-cf95bcca542444d2b21da7a1a1020c612022-12-22T02:14:30ZengBMCJournal of Experimental & Clinical Cancer Research1756-99662011-09-013018310.1186/1756-9966-30-83Effects of metastasis-associated in colon cancer 1 inhibition by small hairpin RNA on ovarian carcinoma OVCAR-3 cellsRen FangWu QinghuaChen ZhiminShi HuirongZhang RuitaoHuang Haoliang<p>Abstract</p> <p>Background</p> <p>Metastasis-associated in colon cancer 1 (MACC1) is demonstrated to be up-regulated in several types of cancer, and can serve as biomarker for cancer invasion and metastasis. To investigate the relations between MACC1 and biological processes of ovarian cancer, MACC1 specific small hairpin RNA (shRNA) expression plasmids were used to investigate the effects of MACC1 inhibition on ovarian carcinoma OVCAR-3 cells.</p> <p>Methods</p> <p>Expressions of MACC1 were detected in different ovarian tissues by immunohistochemistry. MACC1 specific shRNA expression plasmids were constructed and transfected into OVCAR-3 cells. Then, expressions of MACC1 were examined by reverse transcription polymerase chain reaction (RT-PCR) and Western blot. Cell proliferation was observed by MTT and monoplast colony formation assay. Flow cytometry and TUNEL assay were used to measure cell apoptosis. Cell migration was assessed by wound healing and transwell migration assay. Matrigel invasion and xenograft model assay were performed to analyze the potential of cell invasion. Activities of Met, MEK1/2, ERK1/2, Akt, cyclinD1, caspase3 and MMP2 protein were measured by Western blot.</p> <p>Results</p> <p>Overexpressions of MACC1 were detected in ovarian cancer tissues. Expression of MACC1 in OVCAR-3 cells was significantly down-regulated by MACC1 specific small hairpin RNA. In OVCAR-3 cells, down-regulation of MACC1 resulted in significant inhibition of cell proliferation, migration and invasion, meanwhile obvious enhancement of apoptosis. As a consequence of MACC1 knockdown, expressions of Met, p-MEK1/2, p-ERK1/2, cyclinD1 and MMP2 protein decreased, level of cleaved capase3 was increased.</p> <p>Conclusions</p> <p>RNA interference (RNAi) against MACC1 could serve as a promising intervention strategy for gene therapy of ovarian carcinoma, and the antitumor effects of MACC1 knockdown might involve in the inhibition of HGF/Met and MEK/ERK pathways.</p>http://www.jeccr.com/content/30/1/83Ovarian carcinoma OVCAR-3 cellsMetastasis-associated in colon cancer 1Small hairpin RNATherapy target
spellingShingle Ren Fang
Wu Qinghua
Chen Zhimin
Shi Huirong
Zhang Ruitao
Huang Haoliang
Effects of metastasis-associated in colon cancer 1 inhibition by small hairpin RNA on ovarian carcinoma OVCAR-3 cells
Journal of Experimental & Clinical Cancer Research
Ovarian carcinoma OVCAR-3 cells
Metastasis-associated in colon cancer 1
Small hairpin RNA
Therapy target
title Effects of metastasis-associated in colon cancer 1 inhibition by small hairpin RNA on ovarian carcinoma OVCAR-3 cells
title_full Effects of metastasis-associated in colon cancer 1 inhibition by small hairpin RNA on ovarian carcinoma OVCAR-3 cells
title_fullStr Effects of metastasis-associated in colon cancer 1 inhibition by small hairpin RNA on ovarian carcinoma OVCAR-3 cells
title_full_unstemmed Effects of metastasis-associated in colon cancer 1 inhibition by small hairpin RNA on ovarian carcinoma OVCAR-3 cells
title_short Effects of metastasis-associated in colon cancer 1 inhibition by small hairpin RNA on ovarian carcinoma OVCAR-3 cells
title_sort effects of metastasis associated in colon cancer 1 inhibition by small hairpin rna on ovarian carcinoma ovcar 3 cells
topic Ovarian carcinoma OVCAR-3 cells
Metastasis-associated in colon cancer 1
Small hairpin RNA
Therapy target
url http://www.jeccr.com/content/30/1/83
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