Experimental study on the invasiveness inhibition of bladder cancer cells by nuclear factor-κB decoy

来源 :The Chinese-German Journal of Clinical Oncology | 被引量 : 0次 | 上传用户:fcunui_w
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Objective:To determine whether NF-κB is constitutively activated in human bladder cancer cell and,if so,to deter-mine the invasiveness inhibition of bladder cancer cells by nuclear factor-κB decoy—circular dumbbell oligodeoxynucleotides(CD-ODN).Methods:NF-κBp65 activation was determined by immunohistochemical analysis of formalin-fixed,paraffin-embed-ded specimens from 38 cases of bladder transitional cell carcinoma patients.We quantified nuclear staining of RelA as a marker of NF-κBp65 activation.CD-ODN were transfected into human bladder cancer cell line BIU87 by lipofectamine.Luciferase reporter were applied to detecting NF-κB DNA binding activity.The expression levels of uPA were detected by RT-PCR and the cells’ invasion ability by transwell cell culture chamber.Results:P65 excessive activation existed in tumor cell(P<0.01),the activation degree correlated significantly with the expression of uPA(r=0.89,P<0.01),as well as related to tumor invasion-related clinicopathological features such as lymphatic metastasis(P<0.01)and pathological ranking(P<0.05);After transfection with CD-ODN,the activation of NF-κB in BIU87 cell line was suppressed remarkably,the expression level of uPA was decreased and the cells’ invasiveness was weakened as well.Conclusion:Excessively activated NF-κB is related to tumor progression pos-sibly due to its transcriptional regulation of invasion-related factors such as uPA.CD-ODN can efficiently suppress DNA binding activity of NF-κB to reduce the invasive potency of tumor. Objective: To determine whether NF-κB is constitutively activated in human bladder cancer cell and, if so, to deter-mine the invasiveness inhibition of bladder cancer cells by nuclear factor-κB decoy-circular dumbbell oligodeoxynucleotides (CD-ODN). Methods: NF-κBp65 activation was determined by immunohistochemical analysis of formalin-fixed, paraffin-embed-ded specimens from 38 cases of bladder transitional cell carcinoma patients. We quantified nuclear staining of RelA as a marker of NF-κBp65 activation. CD-ODN were transfected into human bladder cancer cell line BIU87 by lipofectamine. Luciferase reporter were applied to detect NF-κB DNA binding activity. The expression levels of uPA were detected by RT-PCR and the cells’ invasion ability by transwell cell culture chamber. Results: P65 excessive activation of in tumor cells (P <0.01), the activation degree correlated significantly with the expression of uPA (r = 0.89, P <0.01), as well as related to tumor invasion-related clinicopathologica After transfection with CD-ODN, the activation of NF-κB was significantly decreased in BIU87 cell line (P <0.01) and pathological ranking cells’ invasiveness was weakened as well. Conlusion: Excessively activated NF-κB is related to tumor progression pos-sibly due to its transcriptional regulation of invasion-related factors such as uPA.CD-ODN can be able to suppress DNA binding activity of NF-κB to reduce the invasive potency of tumor.
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