Combined With Bioinformatics To Analyze The Effect Of Rab23 On Proliferation And Apoptosis Of Gastric Cancer Cells

Objective:The incidence of gastric cancer is high in our country.Due to the lack of attention to the preliminary screening of gastric cancer,most patients with gastric cancer are in the middle or advanced stage,thus missing the best treatment time.Therefore,it is very important to study the mechanism of occurrence,development and metastasis of gastric cancer.As a tumor-related factor,Rab23 plays a key role in tumor proliferation,apoptosis and invasion.The targeting study of mi RNA and Rab23 provides a new idea for the mechanism of tumorigenesis and development.This study was a preliminary investigation of the role of Rab23 in the occurrence and progression of gastric cancer.DEm RNAs and DEmi RNAs were analyzed by bioinformatics,establishing mi RNA-m RNA regulatory network and further searching for potential regulatory mi RNAs of Rab23.The effect of Rab23 on proliferation and apoptosis of gastric adenocarcinoma cells was preliminarily verified in vitro.Methods:1.Detection of Rab23 expression in gastric cancer and paracancerous tissues by q PCR.2.Bioinformatics method was used to integrate and analyze the published m RNA and mi RNA high-throughput transcriptome data of gastric adenocarcinoma.3.Construction of protein-protein interaction(PPI)network composed of DEmi RNA and its target genes by Cytoscape software.4.Preliminary screening of DEmi RNA and target genes based on PPI network.5.GO and KEGG analysis of DEmi RNAs and target genes in bioinformationnetwork by WebGestalt.6.After shRNA-Rab23 transfection of MNK-45 cells,CCK8 assay was used to detect the proliferation of MNK-45 cells.Western blot was used to detect the changes of Rab23,Bcl-2,PI3 K,p-PI3 K and Rac1 in MNK-45 cells.Results:1.q PCR detection showed that the expression of Rab23 m RNA in gastric cancer(6.83±5,12)was higher than that in paracancerous tissues.(P < 0.05)2.4586 DEm RNAs(FDR<0.05)and 30 DEmi RNAs(FDR<0.05)were found by integrated analysis.5091 pairs of mi RNA-m RNA targeting pairs were obtained from mi RWalk database,including 2266 differential target genes.3.The PPI network consisted of 586 nodes and 650 edges.According to PPI network,five HUB proteins were identified.including ENO1(degree=69),HDGF(degree =66),CDK4(degree =50),KPNA2(degree =45)and ATP4A(degree =32).4.Mi RNA-130 a and mi RNA-363 are predicted to regulate the expression of Rab23.5.Based on the GO enrichment analysis,cell proliferation(FDR =0),regulation of cell proliferation(FDR=2.85E-12),vesicle-mediated transport(FDR=0.003872877),endoplasmic reticulum(FDR =0),catalytic complex(FDR =5.72E-14),cytoplasmic vesicle(FDR =6.95E-12)and intracellular vesicle(FDR =6.95E-12)were the most significantly enriched GO terms in gastric cancer.The PI3K-Akt signaling pathway(FDR=0.002893618)and ECM-receptor interaction(FDR =0.002893618)were two significantly enriched pathways in gastric cancer.6.The viability of MKN-45 cells with Rab23 knockdown(0.305±0.004)was lower than that of the control group(0.409±0.005).(P<0.05)Rab23 knockdown significantly decreased the expression level of Bcl-2 and Rac1.There was no significant change in the phosphorylation level of PI3 K protein and p-PI3 K.(P<0.05)Conclusions:1.The expression of Rab23 is up-regulated in gastric cancer,suggesting that Rab23 may be related to the development of gastric cancer.2.Mi RNA-130 a and mi RNA-363 are related to the expression of Rab23.3.The knockdown of Rab23 can inhibit gastric cancer cell proliferation and increase apoptosis.PI3K/Akt pathway is the possible mechanism of Rab23.