Original Article
Copyright ©2013 Baishideng Publishing Group Co., Limited. All rights reserved.
World J Gastroenterol. Feb 14, 2013; 19(6): 866-873
Published online Feb 14, 2013. doi: 10.3748/wjg.v19.i6.866
Interference of suppressor of cytokine signaling 3 promotes epithelial-mesenchymal transition in MHCC97H cells
Yuan-Yuan Ji, Zhi-Dong Wang, Zong-Fang Li, Ke Li
Yuan-Yuan Ji, Ke Li, Scientific Research Center, the Second Affiliated Hospital, Xi’an Jiaotong University School of Medicine, Xi’an 710004, Shaanxi Province, China
Zhi-Dong Wang, Zong-Fang Li, Department of General Surgery and Engineering Research Center of Biotherapy and Translation Medicine, Shaanxi Province, the Second Affiliated Hospital, Xi’an Jiaotong University School of Medicine, Xi’an 710004, Shaanxi Province, China
Author contributions: Ji YY and Wang ZD designed the research and contributed equally to this work; Ji YY and Wang ZD performed the research; Ji YY, Wang ZD and Li K contributed to materials and methods; Ji YY and Li K analyzed the data; Ji YY, Wang ZD and Li ZF wrote the manuscript.
Supported by Program for Changjiang Scholars and Innovative Research Team in Universities, PCSIRT No. 1171; National Natural Science Foundation of China, No. 81201925 and No. 81001588; Specialized Research Fund of the Second Affiliated Hospital of Xi’an Jiaotong University School of Medicine of China, No. RC(XM)201108; and the Fundamental Research Funds for the Central Universities, No. 08143048
Correspondence to: Zhi-Dong Wang, MD, PhD, Department of General Surgery and Engineering Research Center of Biotherapy and Translation Medicine, Shaanxi Province, the Second Affiliated Hospital, Xi’an Jiaotong University School of Medicine, No. 157, West 5th Road, Xi’an 710004, Shaanxi Province, China. xawzd@163.com
Telephone: +86-29-87679386 Fax: +86-29-87679386
Received: September 18, 2012
Revised: December 12, 2012
Accepted: December 22, 2012
Published online: February 14, 2013
Processing time: 152 Days and 21.6 Hours
Abstract

AIM: To investigate the role of suppressor of cytokine signaling 3 (SOCS3) silencing in epithelial-mesenchymal transition (EMT) involved in a human hepatocellular carcinoma MHCC97H cell line.

METHODS: MHCC97H cells were transiently transfected with SOCS3 small-interfering RNA (siRNA). Morphological changes of the transfected cells were observed under microscope. Expressions of E-cadherin, Vimentin and α-smooth muscle actin (α-SMA) were identified with immunofluorescence. Furthermore, protein expressions and mRNA levels of characteristic markers of EMT (E-cadherin, Vimentin, α-SMA and Snail) were detected by Western blotting, quantitative real-time polymerase chain reaction. Transforming growth factor-β1 (TGF-β1) levels in the supernatant were measured with enzyme-linked immunosorbent assay.

RESULTS: The transfected cells with SOCS3 siRNA showed a morphological alteration from a typical cobblestone morphology to mesenchymal spindle-shaped and fusiform features. SOCS3 siRNA lessened immunofluorescent expression of E-cadherin, but elicited immunofluorescent expressions of Vimentin and α-SMA in MHCC97H cells. More importantly, compared with the negative control, depletion of SOCS3 resulted in the decrease of the epithelial marker E-cadherin (P < 0.05), and the increase of the mesenchymal markers Vimentin and α-SMA and the transcription factor Snail in MHCC97H cells (P < 0.05). Moreover, compared with the negative control, SOCS3 siRNA evidently enhanced TGF-β1 secretion in MHCC97H cells (200.20 ± 29.02 pg/mL vs 490.20 ± 92.43 pg/mL, P < 0.05).

CONCLUSION: SOCS3 silencing is able to promote EMT in MHCC97H cells via changing the phenotypic characteristics and modulating the characteristic markers.

Keywords: Hepatocellular carcinoma; Epithelial-mesenchymal transition; Suppressor of cytokine signaling; E-cadherin; Snail