TY - JOUR
T1 - Expression and functional characterization of platelet-derived growth factor receptor-like gene
AU - Guo, Feng Jie
AU - Zhang, Wei Jia
AU - Li, Ya Lin
AU - Liu, Yan
AU - Li, Yue Hui
AU - Huang, Jian
AU - Wang, Jia Jia
AU - Xie, Ping Li
AU - Li, Guan Cheng
PY - 2010/3/28
Y1 - 2010/3/28
N2 - 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.
AB - 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.
KW - Colorectal cancer
KW - Immunohistochemistry
KW - Platelet-derived growth factor receptorlike
KW - Prokaryotic expression
KW - Reverse transcription-polymerase chain reaction
UR - http://www.scopus.com/inward/record.url?scp=77950818808&partnerID=8YFLogxK
U2 - 10.3748/wjg.v16.i12.1465
DO - 10.3748/wjg.v16.i12.1465
M3 - Article
C2 - 20333786
AN - SCOPUS:77950818808
SN - 1007-9327
VL - 16
SP - 1465
EP - 1472
JO - World Journal of Gastroenterology
JF - World Journal of Gastroenterology
IS - 12
ER -