Original Article
Copyright ©2010 Baishideng. All rights reserved
World J Gastroenterol. Mar 28, 2010; 16(12): 1465-1472
Published online Mar 28, 2010. doi: 10.3748/wjg.v16.i12.1465
Expression and functional characterization of platelet-derived growth factor receptor-like gene
Feng-Jie Guo, Wei-Jia Zhang, Ya-Lin Li, Yan Liu, Yue-Hui Li, Jian Huang, Jia-Jia Wang, Ping-Li Xie, Guan-Cheng Li
Feng-Jie Guo, Wei-Jia Zhang, Ya-Lin Li, Yan Liu, Yue-Hui Li, Jian Huang, Jia-Jia Wang, Ping-Li Xie, Guan-Cheng Li, Cancer Research Institute, Xiangya Medical School, Central South University, Changsha 410078, Hunan Province, China
Author contributions: Guo FJ and Li GC designed the research and wrote the paper; Zhang WJ, Li YL, Liu Y, Li YH, Huang J, Wang JJ and Xie PL performed the research and analyzed the data.
Supported by The National Natural Science Foundation of China, No. 30672352
Correspondence to: Guan-Cheng Li, Professor, Cancer Research Institute, Xiangya School of Medicine, Central South University, 110 Xiangya Road, Changsha 410078, Hunan Province, China. libsun@163.com
Telephone: +86-731-4805445 Fax: +86-731-2355042
Received: November 14, 2009
Revised: December 19, 2009
Accepted: December 26, 2009
Published online: March 28, 2010
Abstract

AIM: To investigate the role of platelet-derived growth factor receptor-like gene (PDGFRL) in the anti-cancer therapy for colorectal cancers (CRC).

METHODS: PDGFRL mRNA and protein levels were measured by reverse transcription-polymerase chain reaction (RT-PCR) and immunohistochemistry in CRC and colorectal normal tissues. PDGFRL prokaryotic expression vector was carried out in Escherichia coli (E. coli), and purified by immobilized metal affinity chromatography. The effect of PDGFRL protein on CRC HCT-116 cells was detected by 3-(4,5-dimethylthiazolyl-2)-2,5-diphenyltetrazolium bromide (MTT), clone counting, cell cycle, and wound healing assay.

RESULTS: Both RT-PCR and immunohistochemistry showed that the expression of PDGFRL in colorectal normal tissues was higher than in cancer tissues. Recombinant pET22b-PDGFRL prokaryotic expression vector was successfully expressed in E. coli, and the target protein was expressed in the form of inclusion bodies. After purification and refolding, recombinant human PDGFRL (rhPDGFRL) could efficiently inhibit the proliferation and invasion of CRC HCT-116 cells detected by MTT, clone counting and wound healing assay. Moreover, rhPDGFRL arrested HCT-116 cell cycling at the G0/G1 phase.

CONCLUSION: PDGFRL is a potential gene for application in the anti-cancer therapy for CRC.

Keywords: Platelet-derived growth factor receptor-like; Colorectal cancer; Prokaryotic expression; Reverse transcription-polymerase chain reaction; Immunohistochemistry