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Curcumin induces apoptosis in human gastric carcinoma AGS cells and colon carcinoma HT-29 cells through mitochondrial dysfunction and endoplasmic reticulum stress

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Abstract

In the present study, we investigate the effect of curcumin, a major active component isolated from rhizomes of Curcuma longa, on the cytotoxicity of three human carcinoma cell lines (AGS, HT-29 and MGC803) in gastrointestinal tract and a normal gastric epithelial cell line GES-1, and the mechanism of curcumin-induced apoptosis. The results indicated that curcumin inhibited the gastrointestinal carcinoma cell growth in a dose-dependent manner and cytotoxicity was more towards the gastric carcinoma cell AGS and colon carcinoma cell HT-29 compared to normal gastric cell GES-1, and increased externalization of phosphatidylserine residue was observed by Annexin V/PI staining in the two cell lines. Treatment of AGS and HT-29 cells with curcumin enhanced the cleavage of procaspase-3, -7, -8 and -9. Meanwhile, curcumin induced endoplasmic reticulum (ER) stress and mitochondrial dysfunction as evidenced by up-regulation of CCAAT/enhancer binding protein homologous protein (CHOP), phosphorylation of JNK and down-regulation of SERCA2ATPase, release of cytochrome c, decrease of Bcl-2 and reduction of mitochondrial membrane potential in both AGS and HT-29 cells. Overexpression of bax, total JNK, phospho-FADD and total FADD were also observed in curcumin-treated HT-29 cells. Moreover, curcumin decreased cytosolic and ER Ca2+, but increased mitochondrial Ca2+ in the two cell lines. 2-Aminoethoxydiphenyl borate, an antagonist of inositol 1, 4, 5-triphosphate receptor, partly blocked curcumin-induced cytosolic Ca2+ decrease in AGS and HT-29 cells. Additionally, carbonyl cyanide m-chlorophenylhydrazone, an inhibitor of mitochondrial Ca2+ uptake, reversed curcumin-triggered AGS and HT-29 cells growth inhibition. siRNA to CHOP markedly reduced curcumin-induced apoptosis. These results suggest that curcumin can impact on ER stress and mitochondria functional pathways in AGS and HT-29 cells, death receptor pathway was also involved in curcumin-treated HT-29 cells, thus identifying specific well-defined molecular mechanisms that may be targeted by therapeutic strategies.

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Abbreviations

[Ca2+]c :

Cytosolic calcium

[Ca2+]ER :

Endoplasmic reticulum calcium

[Ca2+]Mito :

Mitochondrial calcium

ER:

Endoplasmic reticulum

CHOP:

CCAAT/enhancer binding protein homologous protein

IP3 :

Inositol 1,4,5-triphosphate

CCCP:

Carbonyl cyanide m-chlorophenylhydrazone

ROS:

Reactive oxygen species

MTT:

3-(4,5)-Dimethylthiahiazo(-z-y1)-3,5-diphenytetrazoliumromide

PI:

Propidium iodide

Fura-2 AM:

Fura-2 acetoxymethyl ester

GRP78/BiP:

Glucose-regulated protein 78/Binding immunoglobulin protein

2-APB:

2-Aminoethoxydiphenyl borate

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Acknowledgments

This study was supported by grants from National Science and Technology Major Project of China (2009ZX09311-003); the Opening Project of Shanghai Key Laboratory of Complex Prescription, Shanghai Science and Technology Committee, P.R. China (Number 10DZ2270900); the Project of Shanghai Leading Academic Discipline, Shanghai Education Committee (J50305), P.R. China; and E-Institute of Traditional Chinese Medicine Internal Medicine, Shanghai Municipal Education Commission (E03008), P.R. China.

Conflict of interest

The authors declare that they have no conflict of interest.

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Authors and Affiliations

  1. Institute of Chinese Materia Medica, Shanghai University of Traditional Chinese Medicine, Shanghai, 201203, China

    Aili Cao, Yang Dong, Hailian Shi & Dazheng Wu

  2. Putuo Hospital, Shanghai University of Traditional Chinese Medicine, Shanghai, 200062, China

    Qi Li, Peihao Yin, Li Wang & Dazheng Wu

  3. Institute of Digestive Diseases, Shanghai University of Traditional Chinese Medicine, Shanghai, 200032, China

    Guang Ji & Jianqun Xie

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  1. Aili Cao
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  2. Qi Li
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Correspondence to Jianqun Xie or Dazheng Wu.

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Aili Cao and Qi Li contributed equally to this work.

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Cao, A., Li, Q., Yin, P. et al. Curcumin induces apoptosis in human gastric carcinoma AGS cells and colon carcinoma HT-29 cells through mitochondrial dysfunction and endoplasmic reticulum stress. Apoptosis 18, 1391–1402 (2013). https://doi.org/10.1007/s10495-013-0871-1

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