Study On The Mechanism Of MiR-107 Regulating Radiosensitivity Of Lung Adenocarcinoma H1299 Cells

Objective: To preliminarily analyze the effect and regulatory mechanism of miR-107 on the radiosensitivity of lung adenocarcinoma H1299 cells,and to provide a theoretical basis for follow-up research.Methods: 1.Bioinformatics prediction of the correlation between miR-107 and ZEB1.3.Construct radioresistant H1299R cells;verify the radioresistance of H1299R cells by CCK-8 cell proliferation assay and clone formation assay;q PCR detects the expression of miR-107 and ZEB1 mRNA in H1299R and H1299 cells;Western Blot detects ZEB1,damageRepair protein,and apoptosis protein expression.4.miR-107 mimic transfection experiment up-regulated miR-107 in H1299R cells;CCK-8 and clone formation assays were used to detect the effect of up-regulation of miR-107 on the radiosensitivity of H1299R cells;q PCR detected miR-107 and miR-107 after up-regulation of miR-107.The expression of ZEB1 mRNA;Western Blot detected the expression of ZEB1,ATM,p-ATM,damage repair protein,and apoptosis protein after up-regulation of miR-107,respectively.4.miR-107 inhibitor transfection experiment down-regulated miR-107 in H1299 cells;CCK-8 and clone formation assays were used to detect the effect of down-regulation of miR-107 on the radiosensitivity of H1299 cells;q PCR detected miR-107 and H1299 cells after down-regulation of miR-107 The expression of ZEB1 mRNA;Western Blot detected the expression of ZEB1,ATM,p-ATM,damage repair protein and apoptosis protein after down-regulation of miR-107,respectively.Results: 1.Bioinformatics analysis showed that miR-107 was negatively correlated with ZEB1(P<0.05).2.The proliferation experiment of CCK-8 cells showed that the proliferation ability of H1299R cells was higher than that of H1299 cells after irradiation at a dose of 6 Gy(24h P>0.05;48h P<0.05;72h P<0.05);The 2Gy survival fraction(SF2)of H1299R cells was higher than that of H1299 cells(P<0.05);q PCR results showed that:miR-107 in H1299R cells Low expression in cells(P<0.05),ZEB1 mRNA was highly expressed in H1299R cells(P<0.05);Western Blot assay detected high expression of ZEB1 protein in H1299R cells(P<0.05).After 2Gy irradiation,PARP protein was highly expressed in H1299R cells,and Caspase3 and cle-PARP proteins were lowly expressed in H1299R cells(P<0.05).3.After up-regulating miR-107 in H1299R cells,q PCR results showed that: miR-107 was highly expressed in the miR-107 mimic group,and ZEB1 mRNA was lowly expressed in the miR-107 mimic group(P<0.05).The CCK-8 proliferation assay showed that: After irradiation at a dose of 6Gy,the proliferation ability of the miR-107 mimic group was lower than that of the negative control group(24h P<0.05;48h P<0.05;72h P<0.05);clone formation experiments showed that with the increase of irradiation dose,miR-107 The number of colonies formed in the mimic group was lower than that in the negative control group,and the SF2 in the miR-107 mimic group was lower than that in the negative control group(P<0.05).The 107 mimic groups had a high expression,while ZEB1,ATM,p-ATM,and PARP proteins were lowly expressed in the miR-107 mimic group(P<0.05).4.After down-regulation of miR-107 in H1299 cells,q PCR results showed that ZEB1 mRNA was highly expressed in the miR-107 inhibitor group(P<0.05);CCK-8proliferation experiments showed that: after 6Gy irradiation,the miR-107 inhibitor group proliferated The ability of miR-107 inhibitor group was higher than that of the negative control group(24h P<0.05;48h P<0.05;72h P<0.05);the colony formation experiment showed that: with the increase of irradiation dose,the number of colonies formed in the miR-107 inhibitor group was higher than that in the negative control group,The SF2 in the miR-107 inhibitor group was greater than that in the negative control group(P<0.05);Western Blot experiments showed that after 2Gy dose irradiation,compared with the negative control group,the expression of caspase3 protein was lower in the miR-107 inhibitor group,ZEB1,p-ATM and PARP protein was highly expressed in the miR-107 inhibitor group(P<0.05).Conclusion: 1.miR-107 can regulate the radiosensitivity of lung adenocarcinoma H1299 cells.2.Ionizing radiation may mediate the apoptosis of H1299 cells through the miR-107/ZEB1/ATM signaling axis.