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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 modulates the MDR phenotype in a dose- and time-dependent manner. (A&B) CCK-8 assay was used to detect the cellular inhibition of L-OHP in HCT116 cells and HCT116/L-OHP cells treated with ZJW at 50 μg/mL for 24, 48, and 72 h. (C) Flow cytometry analysis of apoptosis with Annexin V-FITC/PI binding to HCT116/L-OHP cells after treatment with ZJW at 0, 12.5, 25, and 50 μg/mL for 48 h. (D) Flow cytometry analysis was used to distinguish cells in different phases of the cell cycle as Methods described.
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Fig1: ZJW modulates the MDR phenotype in a dose- and time-dependent manner. (A&B) CCK-8 assay was used to detect the cellular inhibition of L-OHP in HCT116 cells and HCT116/L-OHP cells treated with ZJW at 50 μg/mL for 24, 48, and 72 h. (C) Flow cytometry analysis of apoptosis with Annexin V-FITC/PI binding to HCT116/L-OHP cells after treatment with ZJW at 0, 12.5, 25, and 50 μg/mL for 48 h. (D) Flow cytometry analysis was used to distinguish cells in different phases of the cell cycle as Methods described.

Mentions: We previously showed that ZJW enhances the inhibition rate of chemotherapeutic agents in a dose-dependent manner in MDR tumor cells, including colorectal cancer cells (HCT116/L-OHP), gastric carcinoma cells (SGC7901/DDP), and hepatic carcinoma cells (Bel/Fu). To further examine the effect of ZJW in reversing chemoresistance, we detected the cell proliferation in response to chemotherapeutic agents by the CCK-8 assay and apoptosis by flow cytometry. As shown in Figure 1A, ZJW caused a significant decrease in the cell inhibition rate of chemotherapeutic agents in a time-dependent manner. However, sensitive cell proliferation treatment with ZJW and L-OHP were not found to be significantly different from L-OHP control group (Figure 1B). Similar to our previous study, ZJW increased L-OHP-induced cell apoptosis in a dose-dependent manner (Figure 1C). To further examine the effect of ZJW in cell cycle, Flow cytometric analysis was used after HCT116/L-OHP cells in response to ZJW only without HCT116/L-OHP. However, the cell cycle analyses data showed that there was no change in any phase arrest, especially in sub-G0 population of cell (Figure 1D), suggesting that ZJW did not alter cell cycle in HCT-116/L-OHP cells. Therefore, these data suggest that ZJW is responsible for reversing chemoresistance in its lowest dosage of the IC10 with a time- and dose-dependent manner.Figure 1


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 modulates the MDR phenotype in a dose- and time-dependent manner. (A&B) CCK-8 assay was used to detect the cellular inhibition of L-OHP in HCT116 cells and HCT116/L-OHP cells treated with ZJW at 50 μg/mL for 24, 48, and 72 h. (C) Flow cytometry analysis of apoptosis with Annexin V-FITC/PI binding to HCT116/L-OHP cells after treatment with ZJW at 0, 12.5, 25, and 50 μg/mL for 48 h. (D) Flow cytometry analysis was used to distinguish cells in different phases of the cell cycle as Methods described.
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4288643&req=5

Fig1: ZJW modulates the MDR phenotype in a dose- and time-dependent manner. (A&B) CCK-8 assay was used to detect the cellular inhibition of L-OHP in HCT116 cells and HCT116/L-OHP cells treated with ZJW at 50 μg/mL for 24, 48, and 72 h. (C) Flow cytometry analysis of apoptosis with Annexin V-FITC/PI binding to HCT116/L-OHP cells after treatment with ZJW at 0, 12.5, 25, and 50 μg/mL for 48 h. (D) Flow cytometry analysis was used to distinguish cells in different phases of the cell cycle as Methods described.
Mentions: We previously showed that ZJW enhances the inhibition rate of chemotherapeutic agents in a dose-dependent manner in MDR tumor cells, including colorectal cancer cells (HCT116/L-OHP), gastric carcinoma cells (SGC7901/DDP), and hepatic carcinoma cells (Bel/Fu). To further examine the effect of ZJW in reversing chemoresistance, we detected the cell proliferation in response to chemotherapeutic agents by the CCK-8 assay and apoptosis by flow cytometry. As shown in Figure 1A, ZJW caused a significant decrease in the cell inhibition rate of chemotherapeutic agents in a time-dependent manner. However, sensitive cell proliferation treatment with ZJW and L-OHP were not found to be significantly different from L-OHP control group (Figure 1B). Similar to our previous study, ZJW increased L-OHP-induced cell apoptosis in a dose-dependent manner (Figure 1C). To further examine the effect of ZJW in cell cycle, Flow cytometric analysis was used after HCT116/L-OHP cells in response to ZJW only without HCT116/L-OHP. However, the cell cycle analyses data showed that there was no change in any phase arrest, especially in sub-G0 population of cell (Figure 1D), suggesting that ZJW did not alter cell cycle in HCT-116/L-OHP cells. Therefore, these data suggest that ZJW is responsible for reversing chemoresistance in its lowest dosage of the IC10 with a time- and dose-dependent manner.Figure 1

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