Effect of Tunicamycin on Bovine Capillary Endothelial Cells

GPT inhibitor Tunicamycin develops ER stress causing anti-angiogenic response in breast tumor microvasculature due to unfolded protein response-mediated apoptosis. cDNA microarray identified 123 and 454 differentially regulated genes with 10 genes overlapping between 3h and 32 h of Tunicamycin treatment. Alg-2 expression is inconsistent but not the Dpms. Evidences support that Tunicamycin completely destroys DPMS activity in capillary endothelial cells without affecting its protein or the mRNA levels but by knocking down the phosphorylation. DPMS’ contribution to developing upr during ER stress has therefore been evaluated because of its inherent regulatory property of GPT. FTIR spectroscopy confirmed protein denaturation. Restablishing the phosphorylation status helps the DPMS regaining its activity. As a result, ER stress is reduced reversing apoptosis and bringing more cells into cycling with normal cellular morphology. Furthermore, differential expression of DPMS in breast tumor microvasculature during Tunicamycin therapy raises its potential as a tumor prognostic marker in the clinic. The experimental design consisted of eight microarray experiments. Cy-3 labeled cRNA control targets were combined with Cy-5 labeled cRNA experimental targets obtained from 3 hrs and 32 hrs samples. The combined Cy-3 and Cy-5 cRNAs targets were used for probe hybridization onto the first, third, fifth, seventh microarray, respectively. Dye swapping was done using different biological replicates for both Cy-5 labeled control targets and the Cy-3 experimental targets (as above) and used for hybridization on the second, fourth, the sixth and the eight microarray respectively. Dye swaps using different biological replicates were done to correct for any dye-bias. Two arrays did not pass quality control and were removed from the experiment: Angiogenesis 3h Replicate 1, Angiogenesis 32h Replicate 2.