Regulation Of PER1 Expression By MiR-29a On Invasion And Migration Of Radiation-induced Lung Adenocarcinoma Cells

Objective:(1)To investigate the expression changes of miR-29a in lung adenocarcinoma cells after radiation.(2)To explore the effects of rhythm gene PER1 and miR-29a on invasion and migration of lung adenocarcinoma cells after radiation.(3)To explore the effect of miR-29a regulation of PER1 expression on invasion and migration of lung adenocarcinoma cells after radiation,and preliminarily reveal the related biological mechanisms of invasion and migration of lung adenocarcinoma cells after radiation.Methods:(1)The expression of the rhythm gene PER1 in A549 cells was induced in vitro,and the cells were collected at different times after X-ray irradiation to detect the expression of miR-29a in the cells.(2)After inducing the expression of PER1,the rhythm gene of A549 cells:1.The expression of PER1 gene was knocked down,and the changes of invasion and migration of A549 cells were detected at different times after 8Gy X-ray irradiation,and the protein expressions of PER1 and MMP-2 were detected.2.Overexpression or inhibition of miR-29a,the changes of invasion and migration ability of A549 cells were detected at different time after X-ray irradiation,and the changes of mRNA expression of miR-29a,PER1 and MMP-2 were detected.The protein expression changes of PER1 and MMP-2 were detected.3.MiR-29a was overexpressed and then transfected into the PER1 overexpressed plasmid to detect the expression level of PER1.The mRNA expressions of miR-29a,PER1 and MMP-2 in each group were detected at different times after 8Gy X-ray irradiation,the changes in cell invasion and migration ability,and the protein expressions of PER1 and MMP-2 were detected.Results:(1)Human lung adenocarcinoma A549 cells were cultured in vitro and tPA was added to induce the expression of rhythm gene PER1.The cells were collected at different times,and the PER1 protein expression and rhythm in the cells were detected by Western blot,and the expression of miR-29a in the cells was detected by Real-time PCR.(2)After stimulating the expression of rhythm genes in cultured A549 cells with tPA:1.Per1-siRNA was transfected into cells to interfere with the expression of the rhythm gene PER1.TPA induced cells and TPA+RNAi cells were irradiated by 8Gy X-ray,and the invasion and migration of A549 cells were detected by Transwell experiment at different times after irradiation.Protein expression of PER1 and MMP-2.2.The miR-29a mimics or miR-29a inhibitor were transfected into the cells to overexpress or inhibit miR-29a,respectively.The cells were irradiated by X-ray,and the changes in the invasion and migration ability of A549 cells were detected at different times after irradiation.The changes of miR-29a,PER1 and MMP-2 mRNA expression.The protein expression changes of PER1 and MMP-2 were detected.3.MiR-29a mimics were transfected into cells to overexpress miR-29a and inhibit the expression of PER1,and the expressions of miR-29a and PER1 in cells were detected.Then,Perl overexpression plasmid was transfected into cells to detect the expression level of Per1.The cells were irradiated with 8Gy X-ray,and the mRNA expressions of miR-29a,PER1 and MMP-2 in each group were detected at different times after irradiation,and the changes in cell invasion and migration ability,as well as the protein expressions of PER1 and MMP-2 were detected.Conclusion(s):(1)Induced by TPA,miR-29a in lung adenocarcinoma cells showed periodic changes,and the period of change was about 24 hours.(2)PER1 and miR-29a have inhibitory effects on invasion and migration of lung adenocarcinoma cells after radiation.(3)The mRNA and protein expressions of miR-29a and PER1 in lung adenocarcinoma cells showed dynamic changes after radiation,which increased firstly and then decreased.(4)By regulating the expression of PER1,miR-29a affected the invasion and migration ability of residual lung adenocarcinoma cells after radiation,presenting a dynamic change,which was firstly weakened and then enhanced.