The Effect And Regulation Of MiR-342-5p In Non-small Cell Lung Cancer Cell A549 And H460

MiRNAs are endogenous non-coding RNA with 22 nucleotides in length,which involved in regulating a variety of cellular processes.Aberrant expression of miRNAs can lead to many diseases,mi RNA shows potential applications in cancer diagnosis,prognosis and treatment.In this paper,we studied the role of miR-342-5p and its regulation were studied in non-small cell lung cancer(NSCLC)cell A549 and H460.1.The expression and effect of miR-342-5p in NSCLC A549 and H460Firstly,we detected miR-342-5p expression in lung cancer cells A549 and H460 by real-time quantitative PCR(qPCR).The result showed that the expression of miR-342-5p in A549 and H460 is lower than that of normal lung epithelial cells HBE.Then,in order to investigate the role of miR-342-5p in A549 and H460,miR-342-5p mimics were used to increase the expression of miR-342-5p.The malignant phenotype of cancer cells were observed after A549 and H460 cells were transfected with miR-342-5p mimics.The results showed that overexpression of miR-342-5p significantly inhibited the proliferation,migration,invasion and colony capacity of A549 and H460.To further confirm the above results,A549 and H460 cells were infected with a lentiviral vector with overexpression of miR-342-5p,and stable overexpression cells were screened by puromycin.The results confirmed that overexpression of miR-342-5p significantly inhibited the malignant phenotype of infected cells.Finally,the H460 cells with miR-342-5p stable over-expression were subcutaneous injected into nude mice,results showed that compared to the control group,miR-342-5p over-expression in H460 inhibited the rate of tumor formation,and decresed the volume and the weight of tumor.The above results indicate that miR-342-5p functions as a tumor suppressor gene in lung cancer cells.2.Detection and interaction of miR-342-5p target geneIn order to investigate how miR-342-5p inhibiting the malignant phenotype of A549 and H460 cells,we predicted the target genes of miR-342-5p by using TargetScan7 software,and detected the expression of each candidate target gene in A549 and H460 by qPCR.Among the candidate target genes,the expression of ELP3,FURIN,and PPARGC1 A genes are significantly higher in A549 and H460 cells than that in HBE cells,meanwhile the expression these genes were obviously reduced in A549 and H460 cells with miR-342-5p overexpression.In addition,results from Luciferase assay showed that PPARGC1 A were demonstrated to be targets of miR-342-5p.To further confirm the crosstalk between miR-342-5p and target gene PPARGC1 A,three small interfering RNA(siRNA)sequence targeting PPARGC1 A gene were synthesized and transfected into A549 and H460,then cell malignant phenotype were observed.Results showed that the proliferation,migration,invasion,and colony formation capacity of A549 and H460 cells were significantly inhibited.In conclusion,this study showed that mi R-342-5p is low expressed in lung cancer cell A549 and H460,over-expression of miR-342-5p decrease the proliferation,migration,invasion and colony capacity of A549 and H460 cells,and inhibited in vivo tumor formation.Results from luciferase reporter assay indicated that PPARGC1 A was a target gene of miR-342-5p,the malignant phenotype of A549 and H460 cells were inhibited by knockdown of PPARGC1 A gene.It is so,miR-342-5p may play a role as a tumor inhibitor by directly targeting PPARGC1 A gene.