Objective:To investigate the expression and significance of ADAM12 in bladder cancer.Methods:Transcriptome data and clinical data of bladder cancer and normal samples from TCGA database were downloaded to analyze the ...Objective:To investigate the expression and significance of ADAM12 in bladder cancer.Methods:Transcriptome data and clinical data of bladder cancer and normal samples from TCGA database were downloaded to analyze the expression level and prognosis of ADAM12 in bladder cancer and normal tissues,and analyze its clinical correlation.Bladder cancer wax block samples were collected to detect the expression level of ADAM12 protein in cancer and adjacent cancer by immunohisto-chemistry.In vitro experiments,the experiments were divided into siNC group and siADAM12 group by transfection interference sequence,and the effect of knockdown ADAM12 on the proliferation and migration of bladder cancer cells was evaluated by CCK8,Transwell and scratch experiments.Western Blot was used to detect the expression level of EMT-related protein in each group of bladder cancer cells.Bioinformatics was used to explore the potential mechanism of ADAM12 in influencing the progression of bladder cancer.Results:ADAM12 expression in bladder cancer tissue is higher than normal tissue(P<0.05).Compared with the low expression group,the prognosis of high-expression ADAM12 was worse(P=0.007),and it was related to tumor stage,depth of invasion and lymph node metastasis(P<0.05).The immunohistochemical results showed that ADAM12 was expressed higher in bladder cancer tissues than in neighboring tissues(P<0.05).The results of in vitro experiments showed that knocking down ADAM12 could inhibit the proliferation and migration of bladder cancer cells compared with the control group.Compared with the NC group,the Western Blot test results showed that the expression of E-cadherin was increased in the siADAM12 group,while the expression of N-cadherin and Vimentin was decreased.Overexpression leads to the opposite.Bioinformatics analysis showed that ADAM12 may be involved in multiple cancer-related signaling pathways and is associated with the infiltration of various immune cell(P<0.05).Conclusion:ADAM12 is highly expressed in bladder cancer tissues and is associated with tumor immune cell infiltration and promotes the migration of bladder cancer cells by regulating EMT,which may be a prognostic marker and biotherapeutic target for bladder cancer.展开更多
Objective Pituitary adenomas(PAs)can adapt an aggressive phenotype by invading adjacent brain structures with rapid cellular proliferation.Previous studies demonstrated that excessive expression of metalloproteases AD...Objective Pituitary adenomas(PAs)can adapt an aggressive phenotype by invading adjacent brain structures with rapid cellular proliferation.Previous studies demonstrated that excessive expression of metalloproteases ADAM12 and MMP-14 is instrumental for the active proliferation and invasiveness of PA cells in vitro and of tumors in vivo.However,the mechanisms regulating ADAM12 and MMP-14 expression in PAs remain unclear.Methods Target gene prediction and transcriptomic profiling of invasive vs.noninvasive human PA samples were performed to identify miRNA species potentially involved in the regulation of ADAM12 and MMP14.For cellular analyses of miRNA functions,two mouse PA cell lines(AtT20 and TtT/GF)were transfected with miR-149-3p and miR-149-5p,respectively.The effects of miR-149(3p and 5p)on expression levels of ADAM12 and MMP14 were determined by Western blotting followed by an analysis of proliferation and colony formation assays,scratch migration assays,and invasion assays.Results A significant downregulation of miRNA-149 was observed in invasive vs.noninvasive PA(0.32 vs.0.09,P<0.0001).In AtT-20 and TtT/GF mouse PAs cells,transfection of mimic miRNA-149(3p and 5p)caused a significantly reduced cell proliferation and matrigel invasion,whilst the effect on cell migration was less pronounced.Both strands of miRNA-149(3p and 5p)markedly reduced protein levels of ADAM12 and MMP-14 by at least 40%in both cell lines.Conclusion This study proved that the invasiveness of PA cells is,at least partly,regulated by miRNA-149-dependent expression of ADAM12 and MMP-14.展开更多
基金Natural Science Foundation of Anhui Province(No.1808085MH293)。
文摘Objective:To investigate the expression and significance of ADAM12 in bladder cancer.Methods:Transcriptome data and clinical data of bladder cancer and normal samples from TCGA database were downloaded to analyze the expression level and prognosis of ADAM12 in bladder cancer and normal tissues,and analyze its clinical correlation.Bladder cancer wax block samples were collected to detect the expression level of ADAM12 protein in cancer and adjacent cancer by immunohisto-chemistry.In vitro experiments,the experiments were divided into siNC group and siADAM12 group by transfection interference sequence,and the effect of knockdown ADAM12 on the proliferation and migration of bladder cancer cells was evaluated by CCK8,Transwell and scratch experiments.Western Blot was used to detect the expression level of EMT-related protein in each group of bladder cancer cells.Bioinformatics was used to explore the potential mechanism of ADAM12 in influencing the progression of bladder cancer.Results:ADAM12 expression in bladder cancer tissue is higher than normal tissue(P<0.05).Compared with the low expression group,the prognosis of high-expression ADAM12 was worse(P=0.007),and it was related to tumor stage,depth of invasion and lymph node metastasis(P<0.05).The immunohistochemical results showed that ADAM12 was expressed higher in bladder cancer tissues than in neighboring tissues(P<0.05).The results of in vitro experiments showed that knocking down ADAM12 could inhibit the proliferation and migration of bladder cancer cells compared with the control group.Compared with the NC group,the Western Blot test results showed that the expression of E-cadherin was increased in the siADAM12 group,while the expression of N-cadherin and Vimentin was decreased.Overexpression leads to the opposite.Bioinformatics analysis showed that ADAM12 may be involved in multiple cancer-related signaling pathways and is associated with the infiltration of various immune cell(P<0.05).Conclusion:ADAM12 is highly expressed in bladder cancer tissues and is associated with tumor immune cell infiltration and promotes the migration of bladder cancer cells by regulating EMT,which may be a prognostic marker and biotherapeutic target for bladder cancer.
基金This research was funded in part by the ERANET PerMed consortium“PerProGlio”(to J.W.B.and A.S.).
文摘Objective Pituitary adenomas(PAs)can adapt an aggressive phenotype by invading adjacent brain structures with rapid cellular proliferation.Previous studies demonstrated that excessive expression of metalloproteases ADAM12 and MMP-14 is instrumental for the active proliferation and invasiveness of PA cells in vitro and of tumors in vivo.However,the mechanisms regulating ADAM12 and MMP-14 expression in PAs remain unclear.Methods Target gene prediction and transcriptomic profiling of invasive vs.noninvasive human PA samples were performed to identify miRNA species potentially involved in the regulation of ADAM12 and MMP14.For cellular analyses of miRNA functions,two mouse PA cell lines(AtT20 and TtT/GF)were transfected with miR-149-3p and miR-149-5p,respectively.The effects of miR-149(3p and 5p)on expression levels of ADAM12 and MMP14 were determined by Western blotting followed by an analysis of proliferation and colony formation assays,scratch migration assays,and invasion assays.Results A significant downregulation of miRNA-149 was observed in invasive vs.noninvasive PA(0.32 vs.0.09,P<0.0001).In AtT-20 and TtT/GF mouse PAs cells,transfection of mimic miRNA-149(3p and 5p)caused a significantly reduced cell proliferation and matrigel invasion,whilst the effect on cell migration was less pronounced.Both strands of miRNA-149(3p and 5p)markedly reduced protein levels of ADAM12 and MMP-14 by at least 40%in both cell lines.Conclusion This study proved that the invasiveness of PA cells is,at least partly,regulated by miRNA-149-dependent expression of ADAM12 and MMP-14.
