Effect Of MicroRNA-29a On IL-13-mediated Transcription Factor YY1 In Invasion And Metastasis Of Lung Cancer And Its Mechanism

Background and objectiveIL-13 is a multi-potent cytokine,widely involved in fibrosis,inflammation and tumor occurrence and development.Recent studies have shown that IL-13 can mediate transcription factor YY1 production by PI3K/AKT pathway in vitro model of pulmonary fibrosis.The aim of this study is to investigate whether IL-13 can also be detected in PI3K/AKT pathway mediates the production of YY1 to promote the proliferation and invasion of tumor cells,and the regulation of miR-29 a on this process.Transcription factor YY1 is a member of the zinc finger transcription factor family,its activation or inactivation can promote tumor development,cell growth,but in the occurrence and development of tumor YY1 function remains to be unclear.Studies have shown that microRNA(mi RNA)is involved in the regulation of tumor cell development,differentiation,apoptosis and proliferation.Where miR-29 a is associated with multiple apoptotic factors in cell apoptosis,and they interact to inhibit tumor cell growth.In this study,the expression of YY1 and the phosphorylation of AKT were observed after exacerbation of lung adenocarcinoma A549 cells with exogenous IL-13 at different concentrations and time.The proliferation,apoptosis and apoptosis of lung adenocarcinoma A549 cells were observed by miR-And to explore the relationship between IL-13,YY1 and miR-29 a,and to determine whether IL-13 was through PI3 K in lung adenocarcinoma cells,and to explore the effect of miR-29 a on IL-13-mediated YY1 expression.AKT pathway-mediated YY1 production,and whether miR-29 a affects the proliferation and invasion and migration of lung adenocarcinoma cells by regulating the expression of YY1.Methods:1.To observe the effect of IL-13 mediated signal transduction of transcription factor YY1 expression.IL-13 cells were treated with qRT-PCR at different concentrations and different time.The expression of YY1 mRNA was detected by immunofluorescence assay.The expression of YY1 protein and the phosphorylation of AKT were detected by immunoblotting.2.To observe the effect of PI3 K / AKT pathway on IL-13-induced YY1 mRNA and protein levels and the proliferation and migration of A549 cells.The experimental group: blank control group,IL-13 stimulation group,LY294002(p-AKT inhibitor)group,IL-13 + LY294002 group.A549 cells reached the rate of 80%,replaced with normal medium or containing the corresponding drug medium,the corresponding time after treatment with qRT-PCR detection of YY1 m RNA expression levels,protein immunoblotting YY1 protein expression and AKT phosphate The degree of.Cell scratches and MTT were used to detect cell migration and proliferation.3.To observe the effect of miR-29 a on the biological function of A549 cells in vitro.The miR-29 a plasmid,miR-29 a inhibitor and miR-29 a negative control were transfected into A549 cells.The expression of miR-29 a was detected by real-time quantitative PCR.The apoptosis of A549 cells was detected by flow cytometry The effect of miR-29 a on the proliferation and invasion of A549 cells was detected by MTT and Transwell chamber method.4.To observe the effect of miR-29 a down-regulation of IL-13-mediated expression of transcription factor YY1 on invasion and migration of A549 cells.The experimental group: blank group,miR-29 a group,IL-13 stimulation group,IL-13 +miR-29 a group.The expression of YY1 mRNA was detected by qRT-PCR,and the protein expression level of YY1 was detected by immunoblotting.The expression of YY1 was detected by qRT-PCR.The expression of YY1 mRNA was detected by immunofluorescence.MTT and Transwell cells were used to detect the proliferation and invasion of A549 cells.Results:1.YY1 and P-AKT could reach the maximum at a certain concentration(40ng /ml)and time(12h)after exogenous IL-13 lung adenocarcinoma cell A549,and promote the proliferation and migration ability of A549 cells.2.LY294002 not only inhibited YY1 mRNA and protein expression in A549 cells,but also inhibited the proliferation and migration of A549 cells.3.Increased expression of miR-29 a inhibits A549 cell proliferation,attenuates cell invasion and accelerates its apoptosis4.MiR-29 a inhibits the expression of YY1 m RNA and protein in A549 cells andinhibits the proliferation and invasion of A549 cells.IL-13 can promote the expression of YY1 mRNA and protein in A549 cells and promote the proliferation and invasion of A549 cells.In the presence of miR-29 a,the expression of YY1 and the proliferation of IL-13 in A549 cells were inhibited to a certain extent.Conclusion:In conclusion,IL-13 can induce the expression of YY1 mRNA and protein in A549 cells under certain concentration and time,which affects the phosphorylation of AKT and accelerate cell proliferation and invasion to a certain extent.PI3 K / AKT blocker LY294002 inhibits IL-13-mediated transcription factor YY1 expression and inhibits proliferation and migration of lung adenocarcinoma A549 cells.MiR-29 a can inhibit the production of YY1 in lung adenocarcinoma A549 cells and inhibit the proliferation and invasion of the cells and accelerate its apoptosis.When both IL-13 and miR-29 a act on A549 cells,the above effects can be neutralized.It is suggested that IL-13 can mediate YY1 production and promote cell proliferation and migration ability through PI3 K / AKT pathway in lung adenocarcinoma A549 cells.MiR-29 a can regulate this process.