Effect Of HES1 Gene Silencing On Proliferation,Migration And Invasion In Non-small Cell Lung Cancer A549 Cells

Lung cancer,the malignant tumor with the highest mortality and morbidity,has become a major public health problem worldwide due to high metastasis rate,high recurrence rate,high mortality rate and low prognosis rate.The occurrence of lung cancer is regulated by multi-gene and multi-signal pathway.Therefore,understanding the pathogenesis and mechanism of lung cancer is of great significance in the clinical treatment of lung cancer.Hairy and enhancer of split homolog-1(HES1)is a member of the transcription factor family of proneural basic helix-loop-helix(bHLH).Studies have shown that HES1 as an oncogene not only abnormally expressed in many solid tumors,but also related to the proliferation and metastasis of various malignant tumor cells.HES1 is also highly expressed in non-small cell lung cancer(NSCLC),but there is no effective study on the effect of HES1 on the proliferation,migration and invasion of NSCLC cells.In this experiment,the expression of HES1 was knocked down by lentivirus plasmid infection,and the effect and mechanism of HES1 on the proliferation,migration and invasion of human NSCLC were discussed.This paper has carried out the following studies:One unrelated shRNA and two shRNAs targeting HES1 gene were synthesized in vitro and inserted into pLKO.1 plasmid to form three shRNA expression vectors.RT-PCR and Western blot screened the most effective shRNA interfering with HES1 and detected the knockdown efficiency of HES1 gene.The activity of A549 cells was detected by MTT.The migration ability of A549 cells were examined by wound healing assay and Transwell migration assay.The invasive ability of A549 cells were detected by Transwell invasion assay.The mRNA expression of E-cadherin,Vimentin,SLUG,ZEB1 and MMP2 were detected by RT-PCR.The protein levels of E-cadherin?Vimentin?SLUG?ZEB1?MMP2?MMP9?PTEN?p-AKT?AKT were detected by Western blot.The results of this study are as follows:(1)The shRNA targeting HES1 gene was successfully constructed.Compared with sh-TRC group,the sh-HES1-2 group targeting HES1 was knocked down about 80%.(2)The proliferation of A549 cells in sh-HES1-2 group was significantly lower than that in sh-TRC group.(3)The migration distance of A549 cells in sh-HES1-2 group was lower than that in sh-TRC group.(4)The number of A549 cells in sh-HES1-2 group passing through Transwell polycarbonate membrane after deformation was less than that in sh-TRC group.(5)The number of A549 cells in sh-HES1-2 group passing through polycarbonate membrane after degradation of matrigel gel was lower than that in sh-TRC group.(6)The mRNA expression of E-cadherin was increased and the mRNA expression of SLUG,Vimentin,MMP2 and ZEB1 were decreased after knockdown of HES1 gene.(7)The protein expression of PTEN and E-cadherin were increased,while the protein expression of Vimentin,SLUG,ZEB1,MMP2 and MMP9 were decreased after knockdown of HES1.It is concluded that knockdown of HES1 gene can inhibit the proliferation,migration and invasion of NSCLC A549 cells.The possible molecular mechanism of HES1 involvement in NSCLC A549 cells is through down-regulation of PI3K/AKT signaling pathway and inhibition of EMT process.Therefore,this paper can provide a theoretical basis for finding an effective target of lung cancer.