论文部分内容阅读
Objective:To detect the effects of Polyporus polysaccharide(PPS),Bacillus Calmette-Guerin (BCG),and their combination on the nuclear factor kappa B(NF-κB)signaling pathway associated-gene expression and investigate the molecular mechanisms of the toxic-reducing effect of PPS in coordination with BCG against bladder cancer.Methods:After T739 cells were treated with PPS,BCG and their combination, the changes in mRNA and protein expression of inhibitor of kappa B kinase beta(IKKβ),NF-κB subunit p65 (NF-κB p65),intracellular adhesion molecule 1(ICAM1)and chemokine(C-c motif)ligand 2(CCL2)in bladder cancer cell line T739 were determined by relative quantitative real-time PCR,Western blot,and flow cytometry (FCM).NF-κB p65 DNA-binding activity in T739 cell was detected by biotinylated probe-ELISA,and NF-κB p65 nuclear expression in T739 cell was observed by immunohistochemistry.Results:Compared with the T739 control group,the mRNA expression of IKBKB(IKKβ),Rel A(NF-κB p65),ICAM1 and CCL2 in T739 cells treated with BCG were increased obviously(Ratio>2.0),as well as the expression of IKKβ,CCL2 and ICAM1 proteins.Meanwhile,NF-κB p65 DNA-binding activity and NF-κB p65 nuclear expression in T739 cells treated with BCG were up-regulated significantly(P<0.05).Compared with the control,the increased expression in T739 cells were simultaneously down-regulated after PPS treatment,except for ICAM1 protein expression.With cells treated with a combination of BCG and PPS,the expression of genes associated with the NF-κB signaling pathway,such as IKBKB,ICAM1 and CCL2,were all down-regulated compared to the BCG group,as well as Rel A mRNA expression,NF-κB p65 DNA-binding activity and NF-κB p65 nuclear expression.Conclusions: PPS could inhibit the over-activation of the NF-κB signaling pathway induced by BCG in bladder cancer cells and accordingly attenuate the adverse reactions to BCG therapy.
Objective: To detect the effects of Polyporus polysaccharide (PPS), Bacillus Calmette-Guerin (BCG), and their combination on the nuclear factor kappa B (NF-κB) signaling pathway associated-gene expression and investigate the molecular mechanisms of the toxic- reducing effect of PPS in coordination with BCG against bladder cancer. Methods: After T739 cells were treated with PPS, BCG and their combination, the changes in mRNA and protein expression of inhibitor of kappa B kinase beta (IKKβ), NF-κB subunit p65 (CCL2) in bladder cancer cell line T739 were determined by relative quantitative real-time PCR, Western blot, and flow cytometry (FCM) . NF-κB p65 DNA-binding activity in T739 cells was detected by biotinylated probe-ELISA, and NF-κB p65 nuclear expression in T739 cells was observed by immunohistochemistry. Results: Compared with the T739 control group, the mRNA expression of IKBKB ( IKKβ), Rel A (NF-κB p65), ICAM1 and CCL2 in T739 cells treated with BCG were increased obviously (Ratio> 2.0), as well as the expression of IKKβ, CCL2 and ICAM1 proteins. Meanwhile, NF-κB p65 nuclear-binding activity and NF- with BCG were up-regulated significantly (P <0.05) .Compared with the control, the increased expression in T739 cells were simultaneously down-regulated after PPS treatment, except for ICAM1 protein expression. With cells treated with a combination of BCG and PPS, the expression of genes associated with the NF-κB signaling pathway, such as IKBKB, ICAM1 and CCL2, were all down-regulated compared to the BCG group, as well as Rel A mRNA expression, NF-κB p65 DNA-binding activity and NF -κB p65 nuclear expression.Conclusions: PPS could inhibit the over-activation of the NF-κB signaling pathway induced by BCG in bladder cancer cells and near attenuate the adverse reactions to BCG therapy.