LncRNA LINC00152 Regulates The Proliferation And Apoptosis Of Lung Cancer Cells Via Mir-16/BCL2L2

With the rapid development of medical science and technology,lung cancer continues to be a serious threat to human health.Although the comprehensive treatments with chemotherapy,radiotherapy,surgical treatment,endocrine therapy and targeted therapy as the core treatments have greatly improved the therapeutic effects of lung cancer.But the five-year survival rate of patients is still very low.Non-small cell lung cancer(NSCLC)is the major type of lung cancer.The proliferation,apoptosis,invasion and migration ability of non-small cell lung cancer cells are very complex biological processes,which have direct or indirect impact on the life quality and survival time of lung cancer patients.Non-coding RNAs are a class of non-coding single-stranded RNAs that are ubiquitous organisms and can specifically interact with target genes,resulting in inhibition of mRNA degradation or translation of target genes,affecting the activity of downstream signaling pathways,and ultimately affecting cells behavior.The expression disorder and dysfunction of LncRNA have important influence on the growth and progress of tumors.LINC00152 is one of the newly discovered tumor-associated factors,but the role and potential molecular mechanism of LINC00152 in lung cancer remains to be further studied.Purposes:The aim of the current study was to evaluate the biological behavior and mechanisms of LINC00152 on non-small cell lung cancer,and to provide a theoretical basis for the diagnosis and treatments of non-small cell lung cancer.Methods:1.In vitro,the expression LINC00152 in different non-small cell lung cancer cell lines and normal lung epithelial cells were detected by RT-qPCR,and the most representative lung cancer cell(A549)was screened out.A549 cell was transfected using LINC00152-siRNA.The transfection efficiency was verified by RT-qPCR,CCK-8,flow cytometry were used to detect the proliferation,apoptosis of A549 cells,respectively.2.In order to further explore the underlying mechanisms of LINC00152 in NSCLC,we predicted the potential target genes of LINC00152 through bioinformatics and constructed a fluorescent reporter plasmid.Luciferase reporter assay was used to confirm the relationship between LINC00152 and target gene.The expression of the target gene in non-small cell lung cancer cells was detected by RT-qPCR,western blot and immunohistochemical assay.The expression of the target gene and related factors Caspase 3,Bax,Bcl2,BCL2L2 were detected after overexpressing the expression of LINC00152.The mechanisms of LINC00152 regulating the proliferation and apoptosis of A549 were analyzed.Results:1.Compared to the normal lung epithelial cell,the expression of LINC00152 was decreased in A549 cells(P<0.01).The transfection of LINC00152-siRNA downregulated the LINC00152 expression(P<0.01).Compared with the control and NC groups,the proliferation(P<0.01)of A549 cells were significantly reduced in the LINC00152-siRNA group but the apoptosis rates were significantly increased(P<0.01).2.LINC00152 was validated as a direct target of miR-16 by bioinformatics.And BCL2L2 was validated as a direct target of miR-16 by bioinformatics too.Mir-16 was lowly expressed in NSCLC tissues,and contrary to LINC00152 expression pattern.And BCL2L2 was highly expressed in NSCLC tissues,and contrary to miR-16 expression pattern.LINC00152 down-expression inhibited the expression of Bcl2(P<0.01),BCL2L2(P<0.01)but promoted Caspase3 and Bax(P<0.01).Conclusion:1.LINC00152 is significantly down-regulated in non-small cell lung cancer cell lines.Down-regulation of LINC00152 can induce apoptosis of A549 lung cancer cells and inhibit cell proliferation of A549 cells.It showed that LINC00152 may played a role in promoting cancer in the process of non-small cell lung cancer.2.LINC00152 was a target gene of miR-16,and miR-16 is capable of targeting BCL2L2.Inhibition of expression of LINC00152 can promote the apoptosis-related factors Caspase3 and Bax and inhibit the expression of Bc 12.These results indicated that LINC00152 could regulate the expression of BCL2L2,Caspase3,Bcl2 and Bax through miR-16 and affect the proliferation and apoptosis of A549 cell.