Study On The Mechanism Of Long Non-coding RNA-HCG18 Promoting The Development Of Lung Adenocarcinoma

Objective: To investigate the mechanism of long non-coding RNA-HCG18(lncRNA-HCG18)in lung cancer.Methods:(1)By lung tissue deep sequencing,expression cDNA microarray screening of differentially expressed genes;Analysis paracancerous carcinomas TCGA database lung tissues differentially expressed bioinformatics software;real-time quantitative RT-PCR,100 pairs of clinical lung cancer differential gene expression levels in tissue cancer and adjacent to cancer.(2)Cell functional verification: cck-8 method was used to detect cell proliferation;flow cytometry was used to detect cell cycle;cell scratch test was used to detect cell migration ability;Transwell test was used to determine cell invasion ability.Nude mouse tumorigenesis experiment to detect the effect of knocking down the target gene on the tumorigenic ability of lung cancer cells;(3)Use the transcription factor target prediction database Starbase database to predict the HCG18 binding site and detect the gene binding site by luciferase reporter gene;nuclear-plasma separation experiment to determine the subcellular functional location of the target gene;Western Blot technology and immunohistochemical method verification Functional mechanism of target gene.Result:(1)The results of microarray expression profiling showed that lncRNA-HCG18 was highly expressed in lung cancer(p <0.05);clinical samples and TCGA showed that the expression level of lncRNA-HCG18 cancer tissue was higher than that of adjacent tissue(p <0.05)(2)Cell function experiments verify that lncRNA-HCG18 plays the role of oncogene.The results of cck8 method showed that knocking down lncRNA-HCG18 inhibited the proliferation of cells(p <0.05);Tranwell test results showed that knocking downlncRNA-HCG18 inhibited cells Invasive ability(p <0.05);cell scratch test results showed that knocking down lncRNA-HCG18 can inhibit the migration ability of lung cancer cells(p <0.05);PI staining flow cytometry results showed that lncRNA-HCG18 was not associated with cell cycle,The difference was not statistically significant(p> 0.05);the results of tumor formation in nude mice showed that the tumor growth diameter decreased after knocking down the expression of lncRNA-HCG18(p <0.05);(3)Nuclear-plasma separation experiment results show that lncRNA-HCG18 is located in the cytoplasm;the software predicts that lncRNA-HCG18 has binding sites with has-miR195-5P and ATG14;dual luciferase reporter gene experiment,qPCR,western blot and immunohistochemistry The method showed that the low expression of ATG14 in tumor tissues after the knockdown of lncRNA-HCG18 was detected(p<0.05),indicating that lncRNA-HCG18 can regulate the miR-195 / ATG14 pathway and promote the development of lung cancer.Conclusion: lncRNA-HCG18 is highly expressed in lung adenocarcinoma.Functional experiments have confirmed that lncRNA-HCG18 can promote malignant phenotypes such as proliferation,invasion and metastasis in vivo and in vivo.Mechanism experiments confirmed that lncRNA-HCG18 can affect the activity of miR-195-5p through the "sponge" effect of miRNAs,and competitively combine with ATG14 to form miR-195-5p to form ceRNA model.It leads to increased expression of ATG14,promotes autophagy,and then promotes the development of lung adenocarcinoma.