Downregulation of the G protein-coupled receptor 137 inhibits the proliferation, colony-forming ability and cell cycle distribution of U251 cells

Abstract

Objective To investigate the effect of G protein-coupled receptor 137 (GPR137) on proliferation,colony and cell cycle of glioma cell line U251.Methods Expression levels of GPR137 mRNA in U251,U-87 glioma cells were detected by semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR) ; we used lentivirus-mediated small interfering RNAs (siRNAs) to knock down GPR137 expression in U251 cells.Expression levels of GPR137 mRNA and protein were determined by Real-time PCR and Western blotting; cell proliferation was examined by methyl thiazol tetrazolium (MTT) assay; the colony-forming capacity was verified by colony formation assay; the cell cycle distribution was detected by flow cytometric analysis.Results We found that GPR137 mRNA was highly expressed in U251 and U-87 cells [increased (66.18 ± 1.80) ％ and (33.82 ± 3.40) ％ respectively].After transfection of Lv-sh GPR137 into U251 cells,as compared with control group,inhibition rate of Lv-sh GPR137 reached 76.5％.Inhibition of GPR137 expression in U251 cells by RNAi significantly impaired cell proliferation by 37.4％,the colony-forming capacity,and significantly arrested the cell cycle in S phase (G0/G1:52.18％,S:41.05％,G2/M:6.77％).Conclusion GPR137 is highly expressed in glioma cell lines.Furthermore,GPR137 play a critical role in cell proliferation,colony formation and cell cycle distribution of U251 cells. Key words: G protein-coupled receptor 137 ; Glioma; Proliferation ; Cell coloning; Cell cycle