期刊
WORLD JOURNAL OF GASTROENTEROLOGY
卷 16, 期 12, 页码 1465-1472出版社
BAISHIDENG PUBLISHING GROUP INC
DOI: 10.3748/wjg.v16.i12.1465
关键词
Platelet-derived growth factor receptor-like; Colorectal cancer; Prokaryotic expression; Reverse transcription-polymerase chain reaction; Immunohistochemistry
资金
- National Natural Science Foundation of China [30672352]
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 (MU), 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. (C) 2010 Baishideng. All rights reserved.
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