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Curcumin Reverses 5-Fluorouracil Resistance by Promoting Human Colon Cancer HCT-8/5-FU Cell Apoptosis and Down-regulating Heat Shock Protein 27 and P-Glycoprotein

  • Wen-ting He
  • Yan-hua Zhu
  • Tong Zhang
  • Patima Abulimiti
  • Fan-ye Zeng
  • Li-ping Zhang
  • Ling-juan Luo
  • Xin-mei Xie
  • Hong-liang Zhang
Original Article
  • 21 Downloads

Abstract

Objective

To investigate the potential mechanisms that curcumin reverses 5-fluorouracil (5-FU) multidrug resistance (MDR).

Methods

Cell growth and the inhibitory rate of curcumin (2–25 μg/mL) and/or 5-FU (0.05–1000 μg/mL) on human colon cancer HCT-8 and HCT-8/5-FU (5-FU-resistant cell line) were determined using cell counting kit-8 (CCK-8) assay. Apoptosis and cell cycle after 5-FU and/or curcumin treatment were detected by flflow cytometry (FCM) and transmission electron microscopy (TEM). The expression of the multidrug resistance related factors p-glycoprotein (P-gp) and heat shock protein 27 (HSP-27) genes and proteins were analyzed by reverse transcription polymerase chain reaction (RT-PCR) and Western blotting (WB), respectively.

Results

The inhibitory rate of curcumin or 5-FU on HCT-8 and HCT-8/5-FU cells proliferation at exponential phase were in a dosedependent manner, HCT-8 cell line was more sensitive to curcumin or 5-FU when compared the inhibitory rate of HCT-8/5-FU. The 50% inhibitory concentration (IC50) of combination 5-FU and curcumin (4.0 μg/mL) in HCT-8/5-FU was calculated as 179.26 μg/mL, with reversal fold of 1.85. Another IC50 of combination 5-FU and curcumin (5.5 μg/mL) in HCT-8/5-FU was calculated as 89.25 μg/mL, with reversal fold of 3.71. Synergistic effect of 5-FU and curcumin on HCT-8 and HCT-8/5-FU cells were found. The cell cycle analysis performed by FCM showed that HCT-8 and HCT-8/5-FU cells mostly accumulated at G0/G1 phase, which suggested a synergistic effect of curcumin and 5-FU to induce apoptosis. FCM analysis found that the percentage of apoptosis of cells treated with curcumin, 5-FU and their combination were significantly increased compared to the control group (P<0.05), and the percentage of apoptosis of the combination groups were slightly higher than other groups (P<0.05). The mRNA levels of P-gp (0.28±0.02) and HSP-27 (0.28±0.09) in HCT-8/5-FU cells treated with combination drugs were lower than cells treated with 5-FU alone (P-gp, 0.48±0.07, P=0.009; HSP-27, 0.57±0.10, P=0.007). The protein levels of P-gp (0.25±0.06) and HSP-27 (0.09±0.02) in HCT-8/5-FU cells treated with combination drugs were decreased when compared to 5-FU alone (P-gp, 0.46±0.02, P=0.005; HSP-27, 0.43±0.01, P=0.000).

Conclusions

Curcumin can inhibit the proliferation of human colon cancer cells. Curcumin has the ability of reversal effects on the multidrug resistance of human colon cancer cells lines HCT-8/5-FU. Down-regulation of P-gp and HSP-27 may be the mechanism of curcumin reversing the drug resistance of HCT-8/5-FU to 5-FU.

Keywords

curcumin Chinese medicine 5-fluorouracil multidrug resistance HCT-8 HCT-8/5-FU colon cancer P-gp HSP-27 

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Notes

Acknowledgement

Special thanks to Urumqi OE Biotech Co. Ltd. for their technical assistances in this study.

Supplementary material

11655_2018_2997_MOESM1_ESM.pdf (720 kb)
Curcumin Reverses 5-Fluorouracil Resistance by Promoting Human Colon Cancer HCT-8/5-FU Cell Apoptosis and Down-regulating HSP-27 and P-gp

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Copyright information

© Chinese Association of the Integration of Traditional and Western Medicine 2018

Authors and Affiliations

  • Wen-ting He
    • 1
  • Yan-hua Zhu
    • 1
  • Tong Zhang
    • 2
  • Patima Abulimiti
    • 1
  • Fan-ye Zeng
    • 1
  • Li-ping Zhang
    • 1
  • Ling-juan Luo
    • 1
  • Xin-mei Xie
    • 1
  • Hong-liang Zhang
    • 1
  1. 1.Second Department of OncologyTraditional Chinese Medicine Hospital of Xinjiang Uyghur Autonomous RegionUyghurChina
  2. 2.Department of Oncology, Xiyuan HospitalChina Academy of Chinese Medical SciencesBeijingChina

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