Mechanism Of Ribosomal Protein L23a(RPL23a) Promoting Metastasis In Non-Small Cell Lung Cancer

Objective:Lung cancer is a malignant tumor with the highest morbidity and mortality in the world.At present,great progress has been made in traditional treatment of lung cancer(surgical radiotherapy and chemotherapy),emerging targeted therapy and immunotherapy,but the five-year survival rate of lung cancer has not been significantly improved.Lung cancer metastasis is the most critical factor for poor prognosis.Therefore,it is of great significance to explore the molecular mechanism of NSCLC metastasis to improve patient prognosis Ribosomal Protein L23a(RPL23a)is an RNA-binding protein located at the large subunit of the ribosome.Studies have found that RPL23 a is upregulated in NSCLC metastases,but the specific mechanism is unclear This study aims to explore the role of RPL23 a in the invasion and metastasis of lung cancer and its potential mechanism,so as to provide a research basis for exploring new therapeutic targets in the early diagnosis and treatment of NSCLC.Methods:1.Based on the basis of previous studies and from the perspective of transcriptomics,gene transcription differences were analyzed in 5 pairs of primary lesions of non-small cell lung cancer(adenocarcinoma)and matched metastatic tissues,and the possible gene RPL23 a related to metastasis was analyzed and screened.2.Public databases were used to analyze the expression characteristics of RPL23 a in NSCLC tissues,and GO analysis was performed on genes significantly associated with RPL23 a.3.Western Blot was used to detect the expression of RPL23 a in NSCLC cells with different metastasis abilities,Transwell migration experiment was conducted to study the invasion and metastasis ability of NSCLC,and preliminary screening of cell lines for subsequent RNA sequencing.4.RPL23 a was knocked down in lung adenocarcinoma cell line H1299 by si-RNA transfection method,and the transfection efficiency at m RNA level was verified by q RTPCR 48 h later.5.Transcriptome sequencing technology was used to detect gene expression profiles in H1299 cell lines before and after RPL23 a gene knockout,and R language DESeq2 was used to analyze differentially expressed genes in RNA-seq data of control group and RPL23 a gene knockout group,and functional and pathway enrichment analysis was used.To screen the potential downstream genes associated with RPL23 a in lung adenocarcinoma metastasis and explore its related regulatory signaling pathways.Results:1.Transcriptome data analysis showed that RPL23 a was highly expressed in pleural fluid metastases of non-small cell lung cancer(adenocarcinoma).2.The analysis results of public database showed that RPL23 a expression in NSCLC tissues increased in lung adenocarcinoma,which was consistent with the previous data of this study.Moreover,the high expression of RPL23 a is correlated with the tumor stage and prognosis of lung adenocarcinoma patients.The overall survival rate of patients with high RPL23 a expression decreased and the prognosis was poor.COX regression analysis suggested that RPL23 a could be used as an independent prognostic factor for lung adenocarcinoma,while the results were not significant in lung squamous cell carcinoma.3.Western Blot and Transwell invasion and migration experiments preliminarily verified that NSCLC cell lines with high RPL23 a expression had stronger migration and invasion ability,and adenocarcinoma cell line H1299 was screened for subsequent RNAseq experiments.4.After si-RNA transfection in lung adenocarcinoma cell line H1299,the expression of RPL23 a decreased,and the interference effect was good.5.Transcriptomic analysis showed that there were 48 up-regulated genes and 46down-regulated genes in H1299 cell line transfected with RPL23 a.Comprehensive GO and KEGG enrichment analysis showed that the gene OAS1 and OAS2 were significantly down-regulated after knockdown of RPL23 a.Conclusion:RPL23a gene is highly expressed in the metastases of non-small cell lung cancer(adenocarcinoma),which may be closely related to the development and metastasis of NSCLC.Increased expression in human lung adenocarcinoma was correlated with overall survival rate and clinical stage.RPL23 a may be involved in the regulation of lung cancer metastasis through the regulation of OSA gene family molecules.This study provides a research basis for further exploration of the molecular mechanism of the occurrence and development of lung cancer.RPL23 a is expected to become a target for predicting the diagnosis and treatment of lung adenocarcinoma.