| Objective Through the analysis of TCGA database and GEO database,the high expression of long non-coding RNA DRAIC(lncRNA DRAIC)in lung adenocarcinoma was found.We explored the effect of lncRNA DRAIC on malignant phenotypes such as proliferation,migration,invasion and apoptosis of lung adenocarcinoma cells,and further carried out in-depth research on its mechanism,hoping to bring a new basis and direction to the treatment of lung adenocarcinoma.Method 1 Quantitative real time fluorescene polymerase chain reaction(q RT-PCR)assay was used to detect the expression differences of lncRNA DRAIC and microRNA let-7i-5p between lung adenocarcinoma cells and human lung normal epithelial cells.2 LncRNA DRAIC knockdown test was performed on A549 and H1299 cells by si RNA transfection method.The effects of lncRNA DRAIC knockdown on cell proliferation,migration,invasion and apoptosis were detected by CCK-8 assay,cloning assay,scratch assay,Transwell assay,flow cytometry and Western blot assay.3 Subcellular localization of lncRNA DRAIC was detected by in situ hybridization.Target genes of lncRNA DRAIC were predicted through bioinformatics websites,and the regulation of lncRNA DRAIC on target genes was analyzed by double luciferase reporting assay and q RT-PCR assay.4Micro RNA let-7i-5p overexpression experiments were performed on A549 and H1299 cells by transfecting let-7i-5p mimics.The effects of microRNA let-7i-5p overexpression on cell proliferation,migration,invasion and apoptosis were detected by CCK-8 assay,cloning assay,scratch assay,Transwell assay,flow cytometry and Western blot assay.5Si RNA-DRAIC and let-7i-5p inhibitor were co-transfected in A549 and H1299 cells.The effects of co-transfected si RNA-DRAIC and let-7i-5p inhibitor on cell proliferation,migration,invasion and apoptosis were detected by CCK-8 assay,cloning assay,scratch assay,Transwell assay,flow cytometry and Western blot assay.Results 1 The expression of lncRNA DRAIC in lung adenocarcinoma tissues was higher than that in paracancer tissues,and the expression of lncRNA DRAIC in cell lines was higher than that in normal lung epithelial cells.After si RNA transfection,the expression of lncRNA DRAIC decreased significantly.2 Compared with the control group,CCK-8 and colony experiments showed that the proliferation ability of the knockdown lncRNA DRAIC group decreased,and the scratch and Transwell experiments showed that the migration and invasion ability of the knockdown lncRNA DRAIC group decreased.Flow cytometry showed that the apoptosis rate of knockdown lncRNA DRAIC group increased.Western blot showed that Caspase-3,Caspase-9 and Bax proteins increased and Bcl-2protein decreased in knockdown lncRNA DRAIC group.The results showed that knockdown lncRNA DRAIC group promoted the expression of apoptosis protein.3 In situ hybridization showed that lncRNA was mainly located in cytoplasm.The double lucifase report experiment showed that the fluorescence intensity of let-7i-5p mimics and DRAICWT co-transfected cells was significantly lower than that of mi R-NC and DRAIC-WT cotransfected group.However,after the co-transfection of let-7i-5p mimics and DRAICMUT,the fluorescence intensity was not statistically significant compared with the mi RNC and DRAIC-MUT co-transfection group,and microRNA let-7i-5p expression in lung adenocarcinoma cells was lower than that in normal lung epithelial cells.After transfection with let-7i-5p mimics,microRNA let-7i-5p expression was significantly increased.4Compared with the control group,CCK-8 and colony formation experiments showed that the proliferation ability of cells in the overexpressed microRNA let-7i-5p group decreased,while scratch and Transwell experiments showed that the migration and invasion ability of cells in the overexpressed microRNA let-7i-5p group decreased.Flow cytometry showed that the apoptosis rate of cells in the overexpressed microRNA let-7i-5p group increased.Western blot analysis showed that Caspase-3,Caspase-9 and Bax proteins in the overexpressed microRNA let-7i-5p group increased,while Bcl-2 protein decreased.The results showed that overexpression of microRNA let-7i-5p group promoted the expression of apoptosis protein.5 Rescue experiments showed that inhibition of microRNA let-7i-5p expression partially reversed the inhibitory ability of knockdown lncRNA DRAIC on proliferation,migration and invasion of lung adenocarcinoma cells,and reversed the promoting ability of knockdown lncRNA DRAIC on apoptosis.Conclusion 1 LncRNA DRAIC is highly expressed in lung adenocarcinoma tissues and cells.Knockdown lncRNA DRAIC can inhibit cell proliferation,migration,invasion and promote cell apoptosis.2 Micro RNA let-7i-5p is underexpressed in lung adenocarcinoma cell lines,and overexpression of microRNA.let-7i-5p can inhibit cell proliferation,migration and invasion,and promote cell apoptosis.3 LncRNA DRAIC inhibits proliferation,migration and invasion of lung adenocarcinoma cells and promotes cell apoptosis by targeting up-regulation of microRNA let-7i-5p.Figure 31;Table 2;Reference 139... |
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