| Objective: Prostate cancer(PCa)is the second most common male cancer affecting Western society.Despite significant progress in exploring prostate cancer biomarkers and treatment strategies,mortality from invasive disease remains high.Therefore,it is necessary to find effective predictors related to prostate cancer progression and metastasis,which may help patients to choose appropriate treatment methods and monitor treatment results.In this study,we used the GEO database to screen for V-set and transmembrane domain containing protein 2-like(VSTM2L)genes that are highly expressed in prostate cancer.To verify the specific expression of VSTM2 L gene in prostate cancer tissues and its correlation with clinical pathological parameters of patients,explore its clinical significance,and provide a further theoretical basis for understanding the pathophysiology,diagnosis and treatment of prostate cancer.Methods:(1)Four prostate cancer high-throughput m RNA expression chips were selected by searching the GEO database,including GSE3325,GSE55945,GSE13131,and GSE106363.GSE3325,GSE13131,GSE106363 overexpression differential gene expression fold is set to more than 4 times,GSE55945 overexpression differential gene expression fold is set to more than 2.5 times.There are 98 differential genes in benign and limited genes in GSE3325,753 differential genes in benign and metastatic genes,112 differential genes in GSE55945,431 differential genes in GSE13131,and 4731 differential genes in GSE106363.Using Fun Rich software to make a Wayne diagram,it was concluded that the VSTM2 L gene is one of the four gene chips over-expressing differentially expressed genes.First,the online UALCAN database was used to analyze the VSTM2 L gene data from TCGA,and to verify its differential expression in normal prostate tissue and prostate cancer tissue,and to analyze whether it was related to the level of prostate cancer Gleason score.Secondly,search ONCOMINE database and GEPIA database to verify whether VSTM2 L gene expression in prostate cancer tissues is higher than that in benign tissues.GEPIA database was used to verify the effect of VSTM2 L gene on the overall survival and disease-free survival of patients with prostate cancer.Finally,the The(HPA,Human Protein Atlas)database was searched to verify whether the VSTM2 L gene is overexpressed in prostate cancer,and to analyze the correlation between the VSTM2 L gene expression level and patient survival.Further analysis of the expression of VSTM2 L gene encoding Q96N03 protein in prostate cancer immunohistochemical samples.GO annotation and KEGG pathway enrichment analysis of 99 genes co-expressed with VSTM2 L screened from databases such as ONCOMINE were performed using DAVID and KOBAS 3.0.(2)VSTM2L gene and clinicopathological correlation of prostate cancer patients were analyzed using online software linkedomics.Download TCGA data to analyze the correlation between VSTM2 L gene expression and clinicopathology of 361 patients with prostate cancer and its effect on patient prognosis.(3)Tissue chip combined with immunohistochemical technology was used to study the expression differences of VSTM2 L protein in 3 normal prostate tissues,2 adjacent cancer tissues and 94 cancer tissues,and further study the relationship between the expression level and the clinicopathological characteristics of patients.Results:(1)GEO data analysis shows that VSTM2 L is highly expressed in prostate cancer tissues and more highly expressed in tissues and cell lines with high malignancy.The results of online database verification are consistent with the results of gene chip screening.UALCAN database analysis showed that the overall expression of VSTM2 L gene in prostate cancer tissue was significantly higher than that of benign prostate tissue(p <0.05)With the increase of Gleason score,the expression of VSTM2 L gene increased.Compared with normal prostate tissues,Gleason scores 6,7,8,9 VSTM2 L gene is highly expressed in prostate cancer tissues and has statistical significance(p <0.05),but there is no statistical difference compared with Gleason10 scores.Analysis of ONCOMINE and GEPIA database showed that the expression of VSTM2 L gene in prostate cancer tissues was higher than that in normal tissues(P﹤0.05),but there was no correlation between the expression level and overall survival and disease-free survival(P> 0.05).The Human Protein Atlas database immunohistochemical results showed that VSTM2 L protein expression was up-regulated in prostate cancer tissue compared to normal prostate tissue.ONCOMINE and other databases screened 99 co-expressed genes with VSTM2 L.GO enrichment analysis and Pathway analysis indicate that it is mainly concentrated in the cell and participates in many biological processes: signal transduction regulation,cell communication regulation,negative regulation of phosphorylation,protein binding,Smad binding,etc.The main pathways are concentrated in AMPK Signal pathway,PI3K-Akt signal pathway,etc.(2)The analysis results of online analysis website Linked Omics showed that there was a positive correlation between VSTM2 L gene expression and tumor tissue purity(r = 0.108,p<0.05),which was not related to other clinical pathological data.Analysis of the downloaded TCGA data showed that the high expression rate of VSTM2 L m RNA in PCa tissues was related to positive lymph node metastasis(P = 0.042)and the number of positive lymph nodes(P = 0.02).VSTM2 L m RNA expression was not related to age,surgical marginal status,T stage,Gleason score,clinical stage,pathological grade and distant metastasis(P> 0.05).Spearman rank correlation coefficient analysis showed that the expression of VSTM2 L m RNA in Prostate cancer tissue was related to lymph node metastasis and the number of positive lymph node biopsies(P <0.05).However,the results also showed that VSTM2 L m RNA expression was not related to age,surgical margin status,T stage,Gleason score,clinical stage,pathological grade,and distant metastasis(P> 0.05).Prognostic analysis of 361 patients showed no significant difference in disease-free survival time between the VSTM2 L high and low expression groups(P = 0.577).(3)Tissue microarray results showed that VSTM2 L stained higher in prostate cancer tissues than normal tissues and adjacent tissues.It showed cytoplasm,brownish yellow and tan staining in cell membranes or part of nuclei,and interstitial staining.The clinical pathological analysis showed that with the increase of Gleason score in prostate cancer,the expression rate of VSTM2 L protein increased(P = 0.005).The high expression rate of VSTM2 L in the poorly differentiated group was 58.9%,and the high expression rate of VSTM2 L in the welldifferentiated group was 26.3%,and the difference was statistically significant(P = 0.011).Spearman correlation analysis showed that VSTM2 L protein expression was correlated with Gleason score,age,and pathological grade in prostate cancer tissues(P <0.05).Conclusion:(1)Compared with normal tissues,VSTM2 L gene is highly expressed in prostate cancer tissue,and its expression level is correlated with prostate cancer malignancy,metastasis,Gleason score(6-9),lymph node metastasis and positive number of lymph nodes.(2)GO analysis and KEGG-Pathway analysis indicate that VSTM2 L and co-expressed genes may affect energy metabolism and signal translation in cancer cells by affecting protein binding,oxygen or extracellular matrix binding.It plays a role in tumor cell proliferation,adhesion,invasion and metastasis.(3)Compared with normal and paracancerous tissues,VSTM2 L protein is highly expressed in prostate cancer tissues,mainly expressed in the cytoplasm,cell membrane,part of the nucleus,and VSTM2 L interstitial reaction is positive in some sections.(4)The expression level of VSTM2 L protein in prostate cancer is related to Gleason score and the degree of glandular pathological differentiation.(5)The results of TCGA data analysis did not show that the level of VSTM2 L gene expression was related to the prognosis of patients. |
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