RT-PCR Array Technology Was Used To Screen The Differential Expression Of EMT-related Genes In Bladder Cancer And Up-regulate The Effect Of SOX10 On Bladder Cancer In Vitro

Objective:Target in this study was aimed to observe the expression of genes related to epithelial-mesenchymal transition in normal bladder tissue and bladder carcinoma,providing further evidence for the subsequent study of bladder cancer recurrence and metastasis.We choose sox10 that has much differential expression as target in this reserch.Methods:Twenty-seven cases of bladder cancer specimens were collected,and normal bladder tissues and bladder cancer tissues were distinguished by frozen section.Then,the expressions of 10 genes of related to EMT in bladder cancer tissues and normal bladder tissues were screened by Cancer Pathway Finder PCR Array produced by QIAGEN company,the ten genes are CDH2?DSP?FOXC2?GSC?KRT14?OCLN?SNAI1?SNAI2?SNAI3?SOX10.2.We select the most differentially expressed genes SOX10 as a target gene,and validate the expression of SOX10 by a plurality of samples.We improve the expression of Sox10 in bladder cancer cell lines J82 by lentiviral vectors,assessment change transwell cell invasion ability;q RT-PCR and western blot detection of transfection efficiency;m RNA and protein expression in q RT-PCR and western blot detection technology of E-cadherin;MTT detect proliferation rate change;scratch test to detect cell migration ability to change.Res?lts:Compared with normal bladder tissues,bladder tissue have 4 up-reg?lated genes,6 down-reg?lated genes,up-reg?lated genes are DSP,GSC,KRT14,OCLN,down-reg?lated genes are CDH2,FOXC2,SNAI1,SNAI2,SNAI3,SOX10,SOX10 in bladder cancer of the biggest difference.Therefore we selected SOX10 as target gene,and its diversity of this fluorescence quantitative RT-PCR validation,verification showed that: SOX10 was lower than that of normal tissues(P <0.05)in bladder cancer.Therefore selected as SOX10 gene of interest,and its diversity of this fluorescence quantitative RT-PCR validation,verification showed that: SOX10 was lower than that of normal tissues(P <0.05)in bladder cancer.2.Experimental group transfected with lentiviral overexpression of SOX10 vector,fluorescent display transfection efficiency was 85%;q RT-PCR and western blot detection of transfection efficiency res?lts show: the experimental group SOX10 m RNA and protein expression significantly up-reg?lated(P <0.05);res?lts of q RT-PCR and western blot detection of EMT markers showed that after lentivirus transfection,m RNA and protein expression of E-cadherin significantly upreg?lated(? <0.05);transwell show invasion of the experimental group were significantly lower(P <0.05);scratch test showed what we found that the control group,the experimental group overexpression of SOX10 migration of bladder cancer cells can be significantly suppressed(P <0.05);MTT assay,continuous monitoring of seven days in the experimental group and the control group changes in number of cells,the number of cells in the experimental group compared with the control group and the 490 nm absorbance values were significantly decreased(P <0.05)Conclusion:1.SOX10 expression in bladder cancer is lower than that of normal bladder tissue2.SOX10 can inhibit bladder cancer cell migration,invasion and proliferation,and inhibition of bladder cancer cells EMT.