Sumei Chen1, Shunchang Jiao1, Youchao Jia1, Yang Li2. 1. Department of Medical Oncology, Chinese PLA General Hospital 28 Fuxing Road, Haidian District, Beijing 100853, China. 2. Municipal Key Laboratory, Division of Cardiology, Chinese PLA General Hospital Life Sciences Building, 28 Fuxing Road, Haidian District, Beijing 100853, China.
Abstract
BACKGROUND: The aim of this study was to evaluate the effects of targeted silencing of forkhead box C1 (FOXC1) gene with small interfering RNA (siRNA) on the proliferation and in vitro migration of human non-small-cell lung carcinoma (NSCLC) A549 and NCIH460 cells, and to explore the molecular mechanism. METHODS: These cells were divided into FOXC1 siRNA groups and negative control groups. RESULTS: Quantitative real-time reverse transcription polymerase chain reaction (qRT-PCR) showed that compared with normal cells and paracancerous tissues, FOXC1 mRNA expressions in NSCLC cells and tissues were significantly higher (P<0.05). qRT-PCR and Western blot showed that FOXC1 siRNA effectively silenced FOXC1 gene expression in NSCLC cells. EdU labeling assay revealed that the proliferative capacity significantly decreased compared with that of normal control group after FOXC1 silencing (P<0.05). Significantly fewer cells in the transfected group migrated than those in negative control group did. After FOXC1 silencing, NSCLC cells were arrested in the G0/G1 phase, which were significantly different from those in negative control group (P<0.05). Compared with negative control group, the expression of cyclin D1 decreased and that of E-cadherin increased. Meanwhile, vimentin and MMP-2 expressions significantly reduced (P<0.05). FOXC1 siRNA effectively silenced FOXC1 gene expressions in NSCLC cells, inhibited their proliferation and invasion, and arrested them in the G0/G1 phase, suggesting that FOXC1 affected proliferation probably by regulating the expression of cell cycle-related protein cyclin D1. CONCLUSION: Silencing FOXC1 may evidently inhibit the migration of these cells by reversing the EMT process through suppressing cadherin, being associated with the expressions of extracellular MMPs.
BACKGROUND: The aim of this study was to evaluate the effects of targeted silencing of forkhead box C1 (FOXC1) gene with small interfering RNA (siRNA) on the proliferation and in vitro migration of human non-small-cell lung carcinoma (NSCLC) A549 and NCIH460 cells, and to explore the molecular mechanism. METHODS: These cells were divided into FOXC1 siRNA groups and negative control groups. RESULTS: Quantitative real-time reverse transcription polymerase chain reaction (qRT-PCR) showed that compared with normal cells and paracancerous tissues, FOXC1 mRNA expressions in NSCLC cells and tissues were significantly higher (P<0.05). qRT-PCR and Western blot showed that FOXC1 siRNA effectively silenced FOXC1 gene expression in NSCLC cells. EdU labeling assay revealed that the proliferative capacity significantly decreased compared with that of normal control group after FOXC1 silencing (P<0.05). Significantly fewer cells in the transfected group migrated than those in negative control group did. After FOXC1 silencing, NSCLC cells were arrested in the G0/G1 phase, which were significantly different from those in negative control group (P<0.05). Compared with negative control group, the expression of cyclin D1 decreased and that of E-cadherin increased. Meanwhile, vimentin and MMP-2 expressions significantly reduced (P<0.05). FOXC1 siRNA effectively silenced FOXC1 gene expressions in NSCLC cells, inhibited their proliferation and invasion, and arrested them in the G0/G1 phase, suggesting that FOXC1 affected proliferation probably by regulating the expression of cell cycle-related protein cyclin D1. CONCLUSION: Silencing FOXC1 may evidently inhibit the migration of these cells by reversing the EMT process through suppressing cadherin, being associated with the expressions of extracellular MMPs.
Authors: Martin Jechlinger; Stefan Grunert; Ido H Tamir; Elzbieta Janda; Susanna Lüdemann; Thomas Waerner; Peter Seither; Andreas Weith; Hartmut Beug; Norbert Kraut Journal: Oncogene Date: 2003-10-16 Impact factor: 9.867
Authors: Partha S Ray; Jinhua Wang; Ying Qu; Myung-Shin Sim; Jaime Shamonki; Sanjay P Bagaria; Xing Ye; Bingya Liu; David Elashoff; Dave S Hoon; Michael A Walter; John W Martens; Andrea L Richardson; Armando E Giuliano; Xiaojiang Cui Journal: Cancer Res Date: 2010-04-20 Impact factor: 12.701