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Summer Research Fellowship Programme of India's Science Academies

Chemically induced ER stress and cell death in HEK 293 cells

Swaroopa Gugulothu

Telangana Social Welfare Residential Degree College for Women, Jagityal. 505327

Dr. Bhanuprakash Reddy

HOD, Biochemistry Division, National Institute of Nutrition (NIN), Tarnaka. 500007.

Abstract

In this report I have focused on the UPR (Unfolded Protein Response) which occurs in the cell. ER stress is the area of my interest. For this study I have used two potential chemical inducers (Thapsigargin, Tunicamycin) of endoplasmic reticulum (ER). Tunicamycin inhibits the N glycosylation by preventing the addition of nascent polypeptides to the oligosaccharide and blocks the protein folding mechanism. HEK 293 cells were used in the study. A concentration of 1 mM of thapsigarin and 5ug of tunicamycin is given to the cells independently and studied for the expression level of ER stress markers and cell death markers with the help of western blot. When compared with the cells with no treatment, TM and TG treated cells showed a greater degree of expression in cell death markers and ER stress markers. From this analysis of western blot results, it is observed that TM and TG increases protein aggregation which leads to ER stress and cell death. The expression level of the ER stress markers such as GRP 78, IRE1alpha and cell death markers such as caspase-3, PARP, bax were seen through western blotting analysis and quantification was done by normalization with beta actin.

Keywords: ER stress and unfolded protein response, tunicamycin, thapsigarin, apotosis, cell death, protein aggregation and misfolded protiens.

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