| Gastric cancer is a common malignant tumor of the digestive tract and has become an important public health problem in China.There is a high incidence of gastric cancer in China,and so far national gastric cancer screening has not yet been achieved.Surgery is the main treatment method for gastric cancer,and laparoscopic surgery is the most widely used minimally invasive technique for the treatment of gastric cancer.Most patients with gastric cancer are already advanced at the time of initial consultation,and often relapse after surgical treatment,the 5-year survival rate of patients does not exceed 30%.If accurate and effective screening markers are used for the early screening of gastric cancer,the 5-year survival rate of patients with early gastric cancer will be greater than 90%after treatment.Therefore,it is very important to find specific diagnostic markers for gastric cancer and improve the early diagnosis rate of gastric cancer.RBM24(RNA-binding motif 24)is a member of the RNA-binding protein family and one of the important splicing factors.It mainly regulates the expression of target genes by regulating the mRNA stability or alternative splicing.We downloaded the RNA-seq data of gastric cancer tissues and adjacent tissues in the TCGA(The Cancer Genome Atlas)database,analyzed the expression of RBM24 protein in gastric cancer tissues and correlation between RBM24 protein and survival of patients with gastric cancer with the gastric cancer tissue chip.Then observed the effect of RBM24 on the phenotype of SGC-7901 gastric cancer cells.SGC-7901 gastric cancer cells overexpressing RBM24 gene were sequenced to determine the target molecules that interact with RBM24,and the mechanism of RBM24 in the development of gastric cancer was preliminary studied.Downloaded the RNA-seq data of gastric cancer tissues and adjacent tissues from the TCGA database,used Perl script for extraction,integration and ID conversion,and R language to analyze the expression of RBM24 gastric cancer tissues and adjacent tissues.At the same time,immunohistochemistry of RBM24 protein was performed in gastric cancer tissue chip to determine the expression of RBM24 protein in the nucleus and cytoplasm,analyzed the correlation between RBM24 protein and survival time of gastric cancer patients.Constructed a lentiviral vector that overexpressed RBM24,packaged as lentivirus and infected SGC-7901 gastric cancer cells to overexpress the RBM24 gene.The bioinformatics method was used to analyze the RNA-seq results of SGC-7901 cells that overexpressed the RBM24 gene.The effects of RBM24 on the phenotype of gastric cancer cell line SGC-7901 were studied by experiments such as cell proliferation,cell scratching,transwell,and detection of tumor migration-related proteins.The bioinformatics method was used to analyze the RNA-seq results,and then the differential splicing gene transcripts were verified using the Ensembl online website and PCR experiments,the candidate target genes among the differential genes that may be combined with RBM24 were preliminarily explored.Downloaded 407 data of gastric cancer tissues and adjacent tissue samples from the TCGA database,including 375 gastric cancer tissue samples and 32 adjacent cancer tissue samples.Based on the comprehensive analysis,R language was used to analyze the RBM24 in gastric cancer tissues.The expression of RBM24 gene was found to be down-regulated in gastric cancer tissues.At the same time,immunohistochemical experiments were performed on gastric cancer tissue chips,and RBM24 protein in gastric cancer tissue was up-regulated in cytoplasm and down-regulated in nucleus.After RBM24 was overexpressed in SGC-7901 cells,phenotypic observations were performed through cell proliferation,scratch healing experiments and transwell experiments.It was found that its proliferative capacity,scratch healing ability and migration ability were reduced after overexpression of RBM24,and E-cad protein expression was up-regulated.In the RNA-seq,there were 44 differentially expressed genes,of which SERPINB5,EGR1,SERF1A were up-regulated,and had a RBM24 binding site in the 3’UTR,Using qPCR and Western Blot methods to verify RNA-seq sequencing results,it was found that SERPINB5 was overexpressed at both the RNA and protein levels.Seven differential splicing genes were predicted based on the sequencing results,then we verified the predicted differential splicing genes by PCR.As a result,these genes did not have differential splicing.In summary,the expression of RBM24 mRNA is down-regulated in gastric cancer tissues.The expression of RBM24 protein in gastric cancer tissue was up-regulated in cytoplasm and down-regulated in nucleus,which was consistent with the analysis result of TCGA database..Overexpression of RBM24 in gastric cancer cells can reduce the proliferative capacity,scratch healing ability and migration ability of SGC-7901 gastric cancer cells,and E-cad protein expression was up-regulated,indicating that RBM24 may play an important role in the development of gastric cancer.Analysis of RNA-seq results revealed that SERPINB5 gene was up-regulated and had a binding site for RBM24,it may function as a downstream gene of RBM24.These results provide new research directions for revealing the mechanism of RBM24 in gastric cancer,and provide new ideas for the early diagnosis,prognosis and treatment of gastric cancer. |
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