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NSAIDs may regulate EGR-1-mediated induction of reactive oxygen species and non-steroidal anti-inflammatory drug-induced gene (NAG)-1 to initiate intrinsic pathway of apoptosis for the chemoprevention of colorectal cancer

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Abstract

This study aims to investigate the unclear molecular relationship involved in the activation of intrinsic pathway of apoptosis and NSAID-activated gene-1 (NAG-1) induction as a putative target in NSAIDs-mediated chemoprevention of colorectal cancer. Male Sprague-Dawley rats were administered with a colon-specific pro-carcinogen, 1,2-dimethylhydrazine dihydrochloride to achieve the early stages of colorectal cancer. Histopathological examination was performed for the analysis of neoplastic lesions while flow cytometry was performed for the relative quantification of intracellular reactive oxygen species (ROS), differential mitochondrial membrane potential (MMP or ΔΨ M), and apoptotic events. Various target biomolecules were analyzed either for their mRNA or protein expression profiles via RT-PCR and quantitative Real-Time PCR, or Western blotting and immunofluorescence, respectively. Enhanced gene as well as protein expression of pro-apoptotic agents was observed with the daily oral administration of two NSAIDs viz. Sulindac (cyclooxygenase (COX)-non-specific) and Celecoxib (a selective COX-2 inhibitor). A significant increase in early growth response-1 (EGR-1) protein expression and nuclear localization in NSAIDs co-administered animals may have positively regulated the expression of NAG-1 with a significant enhancement of intracellular ROS in turn decreasing the ΔΨ M to initiate apoptosis. In silico molecular docking analysis also showed that Sulindac and Celecoxib can block the active site pocket of B-cell lymphoma-extra large (Bcl-xL, anti-apoptotic transmembrane mitochondrial protein) which could be a putative mechanism followed by these NSAIDs to overcome anti-apoptotic properties of the molecule. NSAIDs-mediated up-regulation of EGR-1 and thereby NAG-1 along with implication of higher ROS load may positively regulate the intrinsic pathway of apoptosis for the chemoprevention of colorectal cancer.

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Acknowledgments

We thank Mrs. Bhupinder Kaur, CSIC, PGIMER, Chandigarh for her help in acquiring FACS data. Financial assistance from the Department of Biotechnology, Govt. of India (BT/PR11516/MED/30/147/2008) is gratefully acknowledged. We are thankful to University Grants Commission (UGC), Govt. of India for providing fellowship to the research student, Mr. Vivek Vaish. We also acknowledge the contributions of Dr. Ravi Kumar, Application Specialist, Schrödinger, India for kind support in getting trial license for Schrödinger Suit 2011 and help in performing the docking studies.

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The authors declare no conflict of interest.

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  1. Department of Biophysics, Panjab University, Chandigarh, 160 014, India

    Vivek Vaish, Honit Piplani, Chandan Rana & Sankar Nath Sanyal

  2. Department of Histopathology, Post Graduate Institute of Medical Education and Research, Chandigarh, 160 012, India

    Kim Vaiphei

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  1. Vivek Vaish
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Correspondence to Sankar Nath Sanyal.

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Vaish, V., Piplani, H., Rana, C. et al. NSAIDs may regulate EGR-1-mediated induction of reactive oxygen species and non-steroidal anti-inflammatory drug-induced gene (NAG)-1 to initiate intrinsic pathway of apoptosis for the chemoprevention of colorectal cancer. Mol Cell Biochem 378, 47–64 (2013). https://doi.org/10.1007/s11010-013-1593-y

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