c-Met-targeted RNA interference inhibits growth and metastasis of glioma U251 cells in vitro.
Angiogenesis plays an essential role in tumor growth and metastasis and is a promising target for cancer therapy. c-Met, a receptor tyrosine kinase, and its ligand, hepatocyte growth factor (HGF), are critical in cellular proliferation, motility, invasion, and angiogenesis. The present study was designed to determine the role of c-Met in growth and metastasis of glioma U251 cells using RNA interference (RNAi) technology in vitro. We constructed three kinds of shRNA expression vectors aiming at the c-Met gene, then transfected them into glioma U251 cells by lipofectamine(TM) 2000. The level of c-Met mRNA was investigated by real-time polymerse chain reaction (RT-PCR). The protein expression of c-Met was observed by immunofluoresence staining and western blotting. U251 cell growth and adherence was detected by methyl thiazole tetrazolium assay. The apoptosis of U251 cells was examined with a flow cytometer. The adherence, invasion, and in vitro angiogenesis assays of U251 cells were done. We got three kinds of c-Met specific shRNA expression vectors which could efficiently inhibit the growth and metastasis of U251 cells and the expression of c-Met in U251 cells. RT-PCR, immunofluoresence staining and western blotting showed that inhibition rate for c-Met expression was up to 90%, 79% and 85%, respectively. The expression of c-Met can be inhibited by RNA interference in U251 cells, which can inhibit the growth and metastasis of U251 cell and induce cell apoptosis. These results indicate that RNAi of c-Met can be an effective antiangiogenic strategy for glioma.
Chu
Sheng-Hua
SH
Department of Neurosurgery, No. 3 People's Hospital Affiliated to Shanghai Jiao Tong University School of Medicine, China.
Feng
Dong-Fu
DF
Zhang
Hong
H
Chen
Er-Tao
ET
Duan
Zhi-Xin
ZX
Li
Xue-Yuan
XY
Li
Jia
J
Ma
Yan-Bin
YB
Zhu
Zhi-An
ZA
Qiu
Jian-Hua
JH
D017209:Apoptosis; D002448:Cell Adhesion; D002455:Cell Division / Q000235:genetics; D045744:Cell Line, Tumor; D017931:DNA Primers; D005434:Flow Cytometry; D005784:Gene Amplification; D005910:Glioma / Q000235:genetics* / Q000473:pathology; D006801:Humans; D009362:Neoplasm Metastasis / Q000235:genetics / Q000517:prevention & control; D009389:Neovascularization, Pathologic / Q000235:genetics; D034622:RNA Interference; D012333:RNA, Messenger / Q000235:genetics; D012334:RNA, Neoplasm / Q000235:genetics; D034741:RNA, Small Interfering / Q000235:genetics*; D020794:Receptor Protein-Tyrosine Kinases / Q000235:genetics*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014162:Transfection
D017931:DNA Primers; D012333:RNA, Messenger; D012334:RNA, Neoplasm; D034741:RNA, Small Interfering; C080835:RON protein; D020794:Receptor Protein-Tyrosine Kinases