Inhibition Of Proliferation And Migration Of Lung Adenocarcinoma Cells By MRPL12 And Immune Correlation Research

Objectives:To evaluate the expression level of mitochondrial ribosomal protein 12(MRPL12)in lung adenocarcinoma,analyze the possible correlation between MRPL12 and the clinicopathological characteristics of patients,and explore the effect of MRPL12 on the biological function of lung adenocarcinoma cells;To explore the possible influence and correlation of MRPL12 on the immune function of lung adenocarcinoma.It provides a theoretical basis for the discovery of new molecular markers,therapeutic targets and related immunotherapy in the diagnosis and treatment of lung adenocarcinoma.1.Lung adenocarcinoma transcriptome data and clinical data were downloaded and collated from TCGA database,and MRPL12 expression in lung adenocarcinoma,correlation of clinicopathological features and prognostic relevance were analyzed using R software,Perl software.Methods:1.Lung adenocarcinoma transcriptome data and clinical data were downloaded and collated from TCGA database,and MRPL12 expression in lung adenocarcinoma,correlation of clinicopathological features and prognostic relevance were analyzed using R software,Perl software.2.The differential expression of MRPL12 in lung adenocarcinoma was verified from MRPL12 transcriptome level,protein level by using in TIMER,UALCAN,HPA database.The prognostic impact of differential expression of MRPL12 in lung adenocarcinoma was verified with the GEPIA database and Kaplan – Meier plotter database.The correlation and prognostic value between the differential expression of MRPL12 and different clinicopathological features of lung adenocarcinoma were further validated using Kaplan – Meier plotter database.3.The relationship between MRPL12 and immune infiltration was investigated using TISCH,TIMER,and TISIDB databases,which were derived from MRPL12 expression in the TME at the single-cell level,correlation between MRPL12 expression and immune infiltration,correlation between MRPL12 expression and immune cell gene signatures,correlation between MRPL12 expression and immune regulators,correlation between MRPL12 expression and chemokines,and prognostic value of immune cell-based MRPL12 in LUAD,The immune correlates of MRPL12 were explored.4.The biological pathways that MRPL12 might be involved in lung adenocarcinoma were investigated using Gene MANIA;Linked Omics database for MRPL12 interactome analysis,go analysis of MRPL12 and related genes and KEGG pathway analysis.5.RT qPCR was used to measure mrpl12 expression at the m RNA level in lung adenocarcinoma cell lines,and differentially expressed cell lines were selected for further cell functional experiments.6.MRPL12 expression in lung adenocarcinoma cells was knocked down by using mrpl12 siRNA and siNC to transiently transfect lung adenocarcinoma cell lines H1299 and SPCA1;RT qPCR and fluorescence microscopy were used to determine the transfection efficiency of the cell lines;Until after successful transfection,CCK-8,scratch,Transwell migration,invasion were used to study how mrpl12 affects the proliferation,migration,invasion of lung adenocarcinoma cells.7.Statistical analyses were performed using R software and its analysis packages,online bioinformatics databases,and SPSS 22.0 software(IBM,Armonk,NY,USA).Kaplan Meier curves were applied to evaluate the relationship between MRPL12 expression and OS.The association between MRPL12 expression and immune infiltrates,immunomodulators and chemokines was assessed by Spearman correlation analysis.To identify prognostic factors,univariate and multivariate Cox regression analyses were performed.Experimental data were expressed as mean ± SD and compared using student’s t-test.Multiple group comparisons were performed using one-way ANOVA followed by Bonferroni’s post hoc test.Each cell experiment was repeated at least 3 times.Statistical significance was set at P < 0.05.Results:1.R software,online bioinformatics database analysis all showed that the transcriptome and protein expression of MRPL12 were abnormally higher in lung adenocarcinoma;High MRPL12 expression suggested worse prognosis in lung adenocarcinoma patients;High MRPL12 expression was equally suggestive of worse prognosis in different subgroups of clinicopathological features in lung adenocarcinoma patients,and MRPL12 might serve as a prognostic factor to predict patient outcome.2.Bioinformatics analysis showed that high expression of MRPL12 in lung adenocarcinoma was associated with low infiltration levels of multiple immune cells.3.RT qPCR experiments showed that MRPL12 was abnormally highly expressed in multiple lung adenocarcinoma cells compared with normal lung tissue cells;Cell functional assays showed that MRPL12 knockdown significantly inhibited LUAD cell proliferation,migration and invasion.Conclusions:1.MRPL12 is abnormally highly expressed in lung adenocarcinoma,and its high expression suggests a worse prognosis in patients with lung adenocarcinoma.2.The proliferation and migration functions of LUAD cells were inhibited after mrpl12 was knocked down.MRPL12 affects lung adenocarcinoma development as Cancer promoting genes.3.High mrpl12 expression was negatively correlated with the level of immune infiltration in lung adenocarcinoma.4.MRPL12 may serve as a novel therapeutic target and prognostic marker in lung adenocarcinoma,and its differential expression may affect the level of immune cell infiltration and associated immunotherapy in lung adenocarcinoma.