Downregulation Of HNRNPDL Expression Suppresses The Malignant Phenotype Of Lung Adenocarcinoma Cells In Vitro

Objective: After β-elemene acted on A549 and NCI-H1299,the differential gene HNRNPDL was screened by gene chip technology,and to investigate the effect of HNRNPDL on the proliferation of A549 and NCI-H1299 cells and on apoptosis of A549 and NCI-H1299 cells,HNRNPDL was silenced using a lentivirus.Method:1.Preliminary experiments were performed by detecting the cell viability of lung adenocarcinoma cell lines treated with β-elemene using the CCK-8 assay,which indirectly reflects the number of viable cells based on their measured absorbance values,and then reflects the killing ability of the drug on the cells reflecting the influence of the drug on the cells.After treating A549 cells according to the drug sensitivity resμlts,the RNA of each group of samples was extracted by Trizol method and then labeled,and then hybridized with Affymetrix human gene expression profiling microarray(a large dataset of lung cancer samples in GEO database)to detect changes in gene expression levels by analyzing the ratio of the fluorescence intensity of each group of samples hybridized with the probe to determine theβ-elemicromene treatment differentially expressed genes in lung adenocarcinoma cells before and after β-elemene treatment;the genes identified and with potential proliferative and inhibitory functions were then screened using a high-content cell functional screening assay(HCS)with cell counting(Celigo).2.Lentivirus-mediated short hairpin RNA(sh RNA)with the ability to knock down HNRNPDL was constructed.Transfection of A549 and H1299 NSCLC cell lines and experiments were divided into transfection group(sh HNRNPDL)and negative control group(sh Ctrl).Results:1.The IC50 of β-elemicene was 375.5μg/ml and 299.3μg/ml after its action on human lung adenocarcinoma A549 and NCI-H1299 cells respectively by CCK-8;while the concentration was relatively stable after IC30=301.3μg/m L action on A549 cells.IC30 was selected as the final concentration of β-elemene in human lung adenocarcinoma A549 and NCI-H1299 cells in the follow-up experiment;2.A total of 721 genes with significant differential expression were screened,including 273 up-regμlated genes and 448 down-regμlated genes.High Connotation Cell Functional screening assay(HCS)was used to compare the effect of decreased gene set on the cell proliferation rate,and the gene with significant proliferation inhibitory phenotype,HNRNPDL,was initially screened among the target genes to be examined;the best interfering gene,sh HNRNPDL-PSC57267,was screened by cell counting method(Celigo)and validated by RNAi;3.The lentivirus with RNA interference sequence of HNRNPDL gene was successfμlly constructed and transfected with lung adenocarcinoma A549 and H1299 cells,and its transfection efficiency was observed under fluorescence microscope,and cells with an 80% transfection efficiency were used for subsequent experiments;The knockdown efficiency of HNRNPDL was verified by Western Blot and PCR,respectively.;4.The results of CCK-8 assay showed that knockdown of HNRNPDL inhibited the proliferation of A549 and H1299 cells.FCM analysis showed that knockdown of HNRNPDL promoted apoptosis of lung adenocarcinoma cells and blocked the cells in G1 phase.In addition,it was found that the ability to form cell clones was inhibited.Scratch assay showed reduced cell migration ability;Conclusion: The differential gene HNRNPDL,screened by gene microarray technology,inhibits the proliferation of lung adenocarcinoma cells and promotes apoptosis after β-elemene action on lung adenocarcinoma cells.Our results provide further insight into the pathogenesis of NSCLC and suggest new therapeutic targets for lung adenocarcinoma.