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. 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. 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. 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.