云芝多糖对卡介苗刺激的膀胱癌细胞核因子 κB 信号通路的调控作用。

The regulatory effects of polyporus polysaccharide on the nuclear factor kappa B signal pathway of bladder cancer cells stimulated by Bacillus Calmette-Guerin.

机构信息

The Central Laboratory, the Second Affiliated Hospital of Guangzhou University of Traditional Chinese Medicine, Guangzhou 510006, China.

出版信息

Chin J Integr Med. 2011 Jul;17(7):531-6. doi: 10.1007/s11655-010-0787-y. Epub 2011 Jul 3.

DOI:10.1007/s11655-010-0787-y

PMID:21725879

Abstract

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.

摘要

目的

检测多孔菌多糖（PPS）、卡介苗（BCG）及其联合应用对核因子 kappa B（NF-κB）信号通路相关基因表达的影响，探讨 PPS 与 BCG 协同减毒作用的分子机制。

方法

T739 细胞经 PPS、BCG 及其联合处理后，采用相对定量实时 PCR、Western blot 和流式细胞术（FCM）检测膀胱癌 T739 细胞系中 IKKβ（IKKβ）、NF-κB 亚单位 p65（NF-κB p65）、细胞间黏附分子 1（ICAM1）和趋化因子（C-c 基序）配体 2（CCL2）mRNA 和蛋白表达的变化。采用生物素标记探针 ELISA 检测 T739 细胞 NF-κB p65 DNA 结合活性，免疫组织化学法观察 T739 细胞 NF-κB p65 核表达。

结果

与 T739 对照组相比，BCG 处理的 T739 细胞 IKBKB（IKKβ）、Rel A（NF-κB p65）、ICAM1 和 CCL2 的 mRNA 表达明显增加（比值>2.0），同时 IKKβ、CCL2 和 ICAM1 蛋白表达也增加。同时，BCG 处理的 T739 细胞 NF-κB p65 DNA 结合活性和 NF-κB p65 核表达明显上调（P<0.05）。与对照组相比，PPS 处理后 T739 细胞的表达同时下调，除了 ICAM1 蛋白表达。与 BCG 和 PPS 联合处理的细胞相比，与 NF-κB 信号通路相关的基因如 IKBKB、ICAM1 和 CCL2 的表达均下调，Rel A mRNA 表达、NF-κB p65 DNA 结合活性和 NF-κB p65 核表达均下调。