Expression Of SPTBN2 Gene In Bladder Cancer And The Effects On Proliferation,Migration,and Invasion Of Bladder Cancer Cell Line 5637

Objective: To investigate the expression of Spectrin Beta,Non-Erythrocytic 2(SPTBN2)in bladder cancer tissues,the correlation with clinical characteristics and its effects on the proliferation,migration,and invasion of bladder cancer cell line 5637.Methods:(1)TCGA database and GSE13507 data set were used to analyze the expression of SPTBN2 in bladder cancer and its correlation with patient prognosis and clinicopathological factors(2)Single gene GSEA analysis and protein interaction analysis were used to study the potential signaling pathways and biological functions of SPTBN2.(3)RNA interfering was used to knock down the expression of SPTBN2 in bladder cancer cell line 5637.The experiment was divided into blank control group(Blank group),negative control group(NC group),siRNA #1group and siRNA #2 group.The experimental groups(siRNA#1 group and siRNA#2 group)were transfected with SPTBN2 siRNA,while the negative control group was transfected with negative siRNA,and the blank control group was added with the same amount of transfection reagents.Real-time quantitative PCR was used to detect the transcriptional level of SPTBN2 in cells,and Western blot was used to measure the protein expression level of SPTBN2 in cells.Cell proliferation was detected by CCK-8 assay.Cell migration and invasion ability were detected by wound healing assay and Transwell invasion assay.Results:(1)Based on the analysis of TCGA bladder cancer data set and GSE13507 data set,the expression level of SPTBN2 in bladder cancer tissues was higher than that in adjacent normal tissues,and patients with high expression of SPTBN2 had worse prognosis,the difference was statistically significant(P < 0.05).The expression level of SPTBN2 in bladder cancer was correlated with T stage(OR = 2.152,95% CI = 1.390-3.364,P < 0.001),N stage(OR = 1.935,95% CI = 1.257-2.999,P = 0.003)and histological type(OR = 10.313,95%CI = 2.941-65.289,P = 0.002).(2)Single gene GSEA enrichment analysis showed that SPTBN2-related differential genes in bladder cancer were mainly enriched in KRAS signaling pathway.KEGG enrichment analysis showed that SPTBN2-related interacting proteins were mainly enriched in apoptosis,proteoglycans in cancer,mitophagy-animal,gap junction,and bladder cancer signaling pathways.GO enrichment analysis showed that SPTBN2-related interacting proteins were mainly enriched in structural constituent of cytoskeleton,spectrin binding,and ion channel binding in molecular function(MF)category,axon initial segment,main axon,and axon part in the cellular component(CC)category and axon guidance,neuron projection guidance and regulation of membrane potential in the biological process(BP)category.SPTBN2 was associated with genes of MAPK signaling pathway included KRAS(r =0.338,P < 0.001),RAF1(r =0.362,P < 0.001),MAP2K1(r = 0.338,P < 0.001),and MAPK1(r = 0.382,P < 0.001),EGFR(r = 0.360,P < 0.001),the difference was statistically significant.(3)After transfection of SPTBN2 siRNA,the transcription level and protein expression level of SPTBN2 in experimental group were lower than those in negative control group and blank control group.The proliferative,migratory,and invasive abilities of cells in the experimental group were decreased compared with the negative control group,and the difference was statistically significant(P <0.05).Conclusion:(1)The increased expression of SPTBN2 in bladder cancer is correlated with the development of bladder cancer,while high expression of SPTBN2 is related to poor prognosis in patients with bladder cancer.(2)The proliferation,migration,and invasion of bladder cancer cell line 5637 can be inhibited by down-regulating SPTBN2 expression.