Regulatory Role and Mechanism of Tyrosine Kinase Fyn on Proliferation,Migration and Invasion of HepG2 Cells
Objective To reveal the regulatory role of tyrosine kinase Fyn in the proliferation,migration and invasion of HepG2 cells and its possible mechanism.Methods A total of 25 patients diagnosed with hepatocellular carcinoma(HCC)in the Second Affiliated Hospital of Zhengzhou University from January to December 2021 were selected as the study objects.Real-time fluorescence quantitative polymerase chain reaction(RT-qPCR)was used to detect the Fyn expression in HCC tissues and cells.Fyn,signal transducer and activator of transcription 5(STAT5)and NOTCH were down-regulated in HepG2 cells by transfection with the corresponding exogenous siRNAs.The transfected cells were divided into si-Fyn and si-NC groups,and the untreated cells served as the control group.And then the transfection efficiency of HepG2 cells in the three groups was examined by RT-qPCR and protein blotting(Western blotting).Cell counting kit-8(CCK-8)was used to detect cell proliferation.Transwell cell migration assay was used to assess cell migration and invasion ability.And protein blotting was used to detect protein expression of cells and signaling pathway activity.Results The expression of Fyn in HCC tissues and cell lines was higher than that in normal controls(P＜0.05).Fyn silencing significantly inhibited the proliferation,migration and invasion of HepG2 cells,and the expression of related tumor markers CyclinD1,c-Myc,matrix metalloproteinase-2 and matrix metalloproteinase 9(MMP-2/MMP-9)and waveform protein(Vimentin)was decreased,and the expression of p53 was increased(P＜0.05).Further experiments of protein blotting and RT-qPCR showed that Fyn silencing down-regulated the protein levels of p-STAT5 and NICD2 as well as the mRNA expression of NOTCH ligands Jagged-1 and DLL4,and up-regulated the mRNA level of DLL3(P＜0.05),but had no effect on the mRNA expression of Jagged-2 and DLL1(P＞0.05).Meanwhile,STAT5 silencing decreased NICD2 protein level as well as Jagged-1,Jagged-2 and DLL4 mRNA expression(P＜0.05),up-regulated DLL3 mRNA level(P＜0.05),and with no effect on DLL1(P＞0.05).NOTCH2 inactivation inhibited the expression of MMP-2/MMP-9 and Vimentin(P＜0.05).Conclusion Fyn may promote the proliferation,migration and invasion of HepG2 cells by activating the STAT5/NOTCH2 signaling chain.
FynHepatocellular carcinomaSignal transducer and activator of transcription 5NOTCH
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