MiR-140-5p Targeting YES1 Gene Affects Apoptosis And Proliferation Of A549 Cells

MicroRNAs are a class of small(19-24 nucleotides in length),highly conserved non-coding RNAs that may bind to the 3’-untranslated regions(3’-UTRs)of target mRNAs and subsequently suppress the translation of such RNAs to proteins.It has become increasingly evident that different miRNAs may either exhibit oncogenic or tumor-suppressive roles in numerous pathways,depending on their target genes or cellular context.miRNAs are not only involved in the regulation of malignant tumor cell proliferation,but also serve an important role in tumor invasion and metastasis.Therefore,the role of miRNAs in NSCLC has been receiving increasing attention.miRNAs associated with different types of cancer include oncogenic miRNAs,tumor suppressive miRNAs and miRNA regulators.Previous studies have indicated that miR-140-5p serves an important role in numerous tumor types;however,studies investigating the association between miR-140-5p expression and clinical outcomes in patients with resected NSCLC are limited and have demonstrated contradictory results.Therefore,the specific role of miR-140-5p in NSCLC remains to be determined.Objective:In this study,the expression difference of miR-140-5p in lung cancer and adjacent tissues was tested to observe whether its low expression was related to lung cancer.In addition,it was investigated whether miR-140-5p could regulate YES1 proto-oncogene expression,thereby affecting cell proliferation and apoptosis,so as to provide new research ideas for the accumulation data of targeted therapy for lung cancer.Methods:The expression difference of miR-140-5p in lung cancer and adjacent tissues was tested by RT-qPCR to explore whether the low expression of miR-140-5p is related to the occurrence of lung cancer.Lung cancer A549 cell line was cultured in RPMI-1640 complete medium.A549 cells were divided into transfection group,negative control group and NC group,and transfected with miR-140-5p mimics and control mimics respectively.Cells were collected after transfection for 48 h,and apoptosis in each group was detected by flow cytometry.Use TargetScan target genes,such as a database to predict miR-140-5p gene targets,determine YES1 gene is the role of miR-140-5p site,will include targeted point 3 ′ UTR sequences gene synthesis,and set up the negative control group,with Western blot detection,detection YES1,Bcl-2,Bax and Caspase 3 expression of apoptosis related factors such as level,clear whether the miR-140-5p can by acting on YES1 gene caused apoptosis of A549 cells.CCK-8 assay was used to determine whether miR-140-5p could inhibit the proliferation of A549 cells by acting on YES1 gene.Results:The difference of miR-140-5p expression between lung cancer and adjacent tissues was compared by fluorescence PCR,and it was found that the expression level of miR-140-5p in adjacent tissues was much higher than that of lung cancer cell lines(P<0.05).The apoptosis of lung A549 cells transfected with miR-140-5p mimics was detected by flow cytometry,and it was confirmed that the overexpression of miR-140-5p could promote the apoptosis of lung cancer cells.The gene website similar to TargetScan confirms that the possible action sites of miR-140-5p in YES1 from the target gene database,and the 3′utr sequence containing the target sites was synthesized,and the fluorescence reporter vector detection confirmed that YES1 was one of the target gene of miR-140-5p.Western blot was used to investigate the expression of YES1,bcl-2,Bax and caspase-3 in A549 cells transfected with miR-140-5p.The results showed that,compared with the NC group,the ectopic expression of mir-140-5p in A549 cells significantly reduced the expression of YES1 and bcl-2 at mRNA and protein levels,while the expression of Bax and caspase-3 in mimics group was significantly increased.Meanwhile,cck-8 results also confirmed that the cell proliferation ability of mir-140-5p transfected cells was weakened.These results suggest that mir-140-5p may regulate cell viability in vitro by targeting YES1.Conclusion:1.Compared with adjacent tissues,mir-140-5p was significantly reduced in lung cancer tissues.2.Mir-140-5p could target YES1 gene.3.Mir-140-5p affects the apoptosis and proliferation of A549 cells by regulating YES1 gene expression.