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Zuo Jin Wan reverses P-gp-mediated drug-resistance by inhibiting activation of the PI3K/Akt/NF-κB pathway.

Sui H, Pan SF, Feng Y, Jin BH, Liu X, Zhou LH, Hou FG, Wang WH, Fu XL, Han ZF, Ren JL, Shi XL, Zhu HR, Li Q - BMC Complement Altern Med (2014)

Bottom Line: We found that ZJW significantly enhanced the sensitivity of chemotherapeutic drugs and increased oxaliplatin (L-OHP)-induced cell apoptosis in a time- and dose-dependent manner.The effect of ZJW in reversing drug-resistance and suppressing phosphorylation of Akt (Ser473) and NF-κB were weakened after treatment with a PI3K/Akt activator in HCT116/L-OHP cells.Our study has provided the first direct evidence that ZJW reverses drug-resistance in human colorectal cancer by blocking the PI3K/Akt/NF-κB signaling pathway, and could be considered as a useful drug for cancer therapy.

View Article: PubMed Central - PubMed

Affiliation: Department of Medical Oncology, Shuguang Hospital, Shanghai University of Traditional Chinese Medicine, Shanghai 201203, China. Lzwf@hotmail.com.

ABSTRACT

Background: Zuo-Jin-Wan (ZJW), a traditional Chinese medicine formula, has been identified to be effective against drug resistance in cancer. In the present study, we investigated the effect of ZJW on acquired oxaliplatin-resistant and the PI3K/Akt/NF-κB pathway in vitro.

Methods: We tested the dose-response relationship of ZJW on reversing drug-resistance by CCK-8 assay and flow cytometry analysis in vitro. The protein expression of P-gp, MRP-2, LRP, and ABCB1 mRNA expression level were evaluated by Western blot and quantitative RT-PCR. The activities of PI3K/Akt/NF-κB pathway were also examined with or without ZJW, including Akt, IκB, p65 and their phosphorylation expression.

Results: We found that ZJW significantly enhanced the sensitivity of chemotherapeutic drugs and increased oxaliplatin (L-OHP)-induced cell apoptosis in a time- and dose-dependent manner. Moreover, both ZJW and a PI3K specific inhibitor (LY294002) suppressed phosphorylation of Akt (Ser473) and NF-κB, which is necessary in the activation of the PI3K/Akt/NF-κB pathway. The effect of ZJW in reversing drug-resistance and suppressing phosphorylation of Akt (Ser473) and NF-κB were weakened after treatment with a PI3K/Akt activator in HCT116/L-OHP cells.

Conclusions: Our study has provided the first direct evidence that ZJW reverses drug-resistance in human colorectal cancer by blocking the PI3K/Akt/NF-κB signaling pathway, and could be considered as a useful drug for cancer therapy.

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ZJW reversed constitutive as well as PI3K/Akt pathway-induced MDR phenotype. (A) Anti-proliferative IC50 values of L-OHP on HCT116/L-OHP cell treated with LY294002 (20 μM, 2 h), ZJW (50 μg/mL, 48 h), and a combination of ZJW (50 μg/mL, 48 h) and IGF-1 (100 ng/mL, 48 h), were analyzed by CCK-8. Data are presented as mean ± SD of triplicate experiments. **P < 0.01 vs. control, #P < 0.05 vs. ZJW group. (B) Flow cytometry analysis of apoptosis with Annexin V-FITC/PI binding to HCT116/L-OHP cells treated with LY294002 (20 μM, 2 h), ZJW (50 μg/mL, 48 h), and a combination of ZJW (50 μg/mL, 48 h) and IGF-1 (100 ng/mL, 48 h).
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Fig2: ZJW reversed constitutive as well as PI3K/Akt pathway-induced MDR phenotype. (A) Anti-proliferative IC50 values of L-OHP on HCT116/L-OHP cell treated with LY294002 (20 μM, 2 h), ZJW (50 μg/mL, 48 h), and a combination of ZJW (50 μg/mL, 48 h) and IGF-1 (100 ng/mL, 48 h), were analyzed by CCK-8. Data are presented as mean ± SD of triplicate experiments. **P < 0.01 vs. control, #P < 0.05 vs. ZJW group. (B) Flow cytometry analysis of apoptosis with Annexin V-FITC/PI binding to HCT116/L-OHP cells treated with LY294002 (20 μM, 2 h), ZJW (50 μg/mL, 48 h), and a combination of ZJW (50 μg/mL, 48 h) and IGF-1 (100 ng/mL, 48 h).

Mentions: Previous studies have demonstrated that the PI3K/Akt pathway is activated in various tumor MDR cell types. To ascertain whether the reversal of drug-resistance by ZJW in colorectal cancer cells is correlated with the activation of the PI3K/Akt pathway, a PI3K specific inhibitor (LY294002) and activator (IGF-1) were added to HCT116/L-OHP cells as described. As expected, pretreatment with LY294002 or ZJW increased the sensitivity to chemotherapy and allowed the chemotherapeutic drugs to induce cell apoptosis (Figure 2A and B). However, the combination ZJW and IGF-1 weakened the reversal of L-OHP resistance as compared with the ZJW or LY294002 groups (Figure 2A and B).Figure 2


Zuo Jin Wan reverses P-gp-mediated drug-resistance by inhibiting activation of the PI3K/Akt/NF-κB pathway.

Sui H, Pan SF, Feng Y, Jin BH, Liu X, Zhou LH, Hou FG, Wang WH, Fu XL, Han ZF, Ren JL, Shi XL, Zhu HR, Li Q - BMC Complement Altern Med (2014)

ZJW reversed constitutive as well as PI3K/Akt pathway-induced MDR phenotype. (A) Anti-proliferative IC50 values of L-OHP on HCT116/L-OHP cell treated with LY294002 (20 μM, 2 h), ZJW (50 μg/mL, 48 h), and a combination of ZJW (50 μg/mL, 48 h) and IGF-1 (100 ng/mL, 48 h), were analyzed by CCK-8. Data are presented as mean ± SD of triplicate experiments. **P < 0.01 vs. control, #P < 0.05 vs. ZJW group. (B) Flow cytometry analysis of apoptosis with Annexin V-FITC/PI binding to HCT116/L-OHP cells treated with LY294002 (20 μM, 2 h), ZJW (50 μg/mL, 48 h), and a combination of ZJW (50 μg/mL, 48 h) and IGF-1 (100 ng/mL, 48 h).
© Copyright Policy - open-access
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4288643&req=5

Fig2: ZJW reversed constitutive as well as PI3K/Akt pathway-induced MDR phenotype. (A) Anti-proliferative IC50 values of L-OHP on HCT116/L-OHP cell treated with LY294002 (20 μM, 2 h), ZJW (50 μg/mL, 48 h), and a combination of ZJW (50 μg/mL, 48 h) and IGF-1 (100 ng/mL, 48 h), were analyzed by CCK-8. Data are presented as mean ± SD of triplicate experiments. **P < 0.01 vs. control, #P < 0.05 vs. ZJW group. (B) Flow cytometry analysis of apoptosis with Annexin V-FITC/PI binding to HCT116/L-OHP cells treated with LY294002 (20 μM, 2 h), ZJW (50 μg/mL, 48 h), and a combination of ZJW (50 μg/mL, 48 h) and IGF-1 (100 ng/mL, 48 h).
Mentions: Previous studies have demonstrated that the PI3K/Akt pathway is activated in various tumor MDR cell types. To ascertain whether the reversal of drug-resistance by ZJW in colorectal cancer cells is correlated with the activation of the PI3K/Akt pathway, a PI3K specific inhibitor (LY294002) and activator (IGF-1) were added to HCT116/L-OHP cells as described. As expected, pretreatment with LY294002 or ZJW increased the sensitivity to chemotherapy and allowed the chemotherapeutic drugs to induce cell apoptosis (Figure 2A and B). However, the combination ZJW and IGF-1 weakened the reversal of L-OHP resistance as compared with the ZJW or LY294002 groups (Figure 2A and B).Figure 2

Bottom Line: We found that ZJW significantly enhanced the sensitivity of chemotherapeutic drugs and increased oxaliplatin (L-OHP)-induced cell apoptosis in a time- and dose-dependent manner.The effect of ZJW in reversing drug-resistance and suppressing phosphorylation of Akt (Ser473) and NF-κB were weakened after treatment with a PI3K/Akt activator in HCT116/L-OHP cells.Our study has provided the first direct evidence that ZJW reverses drug-resistance in human colorectal cancer by blocking the PI3K/Akt/NF-κB signaling pathway, and could be considered as a useful drug for cancer therapy.

View Article: PubMed Central - PubMed

Affiliation: Department of Medical Oncology, Shuguang Hospital, Shanghai University of Traditional Chinese Medicine, Shanghai 201203, China. Lzwf@hotmail.com.

ABSTRACT

Background: Zuo-Jin-Wan (ZJW), a traditional Chinese medicine formula, has been identified to be effective against drug resistance in cancer. In the present study, we investigated the effect of ZJW on acquired oxaliplatin-resistant and the PI3K/Akt/NF-κB pathway in vitro.

Methods: We tested the dose-response relationship of ZJW on reversing drug-resistance by CCK-8 assay and flow cytometry analysis in vitro. The protein expression of P-gp, MRP-2, LRP, and ABCB1 mRNA expression level were evaluated by Western blot and quantitative RT-PCR. The activities of PI3K/Akt/NF-κB pathway were also examined with or without ZJW, including Akt, IκB, p65 and their phosphorylation expression.

Results: We found that ZJW significantly enhanced the sensitivity of chemotherapeutic drugs and increased oxaliplatin (L-OHP)-induced cell apoptosis in a time- and dose-dependent manner. Moreover, both ZJW and a PI3K specific inhibitor (LY294002) suppressed phosphorylation of Akt (Ser473) and NF-κB, which is necessary in the activation of the PI3K/Akt/NF-κB pathway. The effect of ZJW in reversing drug-resistance and suppressing phosphorylation of Akt (Ser473) and NF-κB were weakened after treatment with a PI3K/Akt activator in HCT116/L-OHP cells.

Conclusions: Our study has provided the first direct evidence that ZJW reverses drug-resistance in human colorectal cancer by blocking the PI3K/Akt/NF-κB signaling pathway, and could be considered as a useful drug for cancer therapy.

Show MeSH
Related in: MedlinePlus