Effects Of LMTK3 Down-regulation On Cell Biological Behaviors Of Lung Squamous Carcinoma Cell Line SK-MES-1 And Its Related Mechanisms

Objective: In this study,effects of down-regulation of lemur tyrosine kinase-3(LMTK3)on proliferation,cell cycle and invasion ability of lung squamous carcinoma cell line SK-MES-1 was investigated and its related mechanisms was explored.Methods: The recombinant of LMTK3-small hairpin RNA(LMTK3-sh RNA)was transfected into SK-MES-1 cells by lentivirus plasmid transfection.After selection of cells with puromycin for two weeks stable cell line SK-MES-1 expressing gene of LMTK3 specific sh RNA was obtained.Cell proliferation affected by LMTK3 was determined by cell counting kit(CCK-8)assay,cell cycle was analyzed by flow cytometry,and cell invasion was detected by transwell assay.LMTK3,?-catenin and cyclin D1 m RNA and protein expression levels were detected by real-time quantitative polymerase chain reaction(Real-Time q PCR)and Western blotting respectively.Results: The results of Real-Time q PCR and Western blot displayed down-regulation of LMTK3 in m RNA and protein level of SK-MES-1 cells transfected with LMTK3-sh RNA.The LMTK3 m RNA relative expression of LMTK3-sh RNA group(0.24�0.18)was significantly lower than that of mock group(1.00�0.18)and NC group(0.93�0.19),(P<0.05).The LMTK3 protein expression level of LMTK3-sh RNA group(0.49�0.07)was significantly lower than that of mock group(1.00�0.11)and NC group(0.99�0.07),(P<0.05).CCK-8 assay revealed that the proliferation of SK-MES-1 cells was significantly inhibited after transfection with LMTK3-sh RNA.Compared with the mock and the NC group,SK-MES-1 cells were blocked in G1 phase(P<0.05)and the number of penetrating the membrane was significantly reduced [(45�5)vs(198�8)and(171�4)cells,P<0.05] after the transfection with LMTK3-sh RNA.And the gene transcription and protein expression of ?-catenin and cyclin D1 in the experimental group were significantly decreased.Conclusion: LMTK3 gene silencing could significantly inhibit the proliferation and invasion of SK-MES-1 cells,blocked the cells in G1 phase,which may be associated with the inhibition of Wnt/?-catenin signaling pathway.