Apoptosis induced by adenosine involves endoplasmic reticulum stress in EC109 cells

Wu,Ling-Fei; Wei,Bi-Liu; Guo,Yi-Tian; Ye,Yan-Qing; Li,Guo-Ping; Pu,Ze-Jin; Feng,Jia-Lin

doi:10.3892/ijmm.2012.1085

Apoptosis induced by adenosine involves endoplasmic reticulum stress in EC109 cells

Authors:
- Ling-Fei Wu
- Bi-Liu Wei
- Yi-Tian Guo
- Yan-Qing Ye
- Guo-Ping Li
- Ze-Jin Pu
- Jia-Lin Feng
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Affiliations: Department of Gastroenterology, The Second Affiliated Hospital, Shantou University Medical College, Shantou 515041, P.R. China, Department of Information, The Second Affiliated Hospital, Shantou University Medical College, Shantou 515041, P.R. China
Published online on: August 2, 2012 https://doi.org/10.3892/ijmm.2012.1085
Pages: 797-804

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Abstract

Apoptosis plays a critical role in the development and homeostasis of multicellular organisms, and endoplasmic reticulum stress (ERS) is one of the intrinsic apoptosis pathways. Previous studies have shown that adenosine induces apoptosis in several cancer cell lines. However, the molecular mechanism remains poorly understood. In this study, we explored whether adenosine triggers apoptosis of EC109 esophageal carcinoma (EC) cells by ERS. The MTT assay was used to determine cell proliferation; cell cycle detection (FCM) and terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) assay were performed to determine cell apoptosis. The subcellular distribution and expression of the ERS-related proteins GRP78, cleaved caspase-3, cleaved caspase-4, CHOP and NF-κB p65 were detected by western blot techniques. NF-κB activation was measured by electrophoretic mobility shift assay (EMSA). The MTT assay demonstrated that adenosine inhibited EC109 cell proliferation in a dose- and time-dependent manner. FCM and TUNEL assay verified that adenosine caused an apoptotic peak in cell cycle arrest and a higher percentage of apoptotic cells. Western blot analysis confirmed that the expression of GRP78, cleaved caspase-4, CHOP, NF-κB p65 and cleaved caspase-3 were upregulated in a dose-dependent manner after adenosine treatment. EMSA revealed that adenosine activated NF-κB p65. This is the first demonstration that adenosine inhibits cell proliferation, increases GRP78 and NF-κB p65 expression and induces apoptosis by CHOP and caspase-4 pathways. The ERS pathway is involved in adenosine-induced apoptosis in EC109 cells.

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