Investigating Hippo-YAP Downstream Gene Expression Under Glutamine and Serum Deprivation in HEp2 Cancer Cells

Abstract

The Hippo-YAP signaling pathway is becoming more and more important in cancer research because of its crucial function in controlling cell growth, proliferation, and apoptosis. By simulating the tumor microenvironment that can be shown by CTPS filament formation, nutrient deprivation is theorized to affect Yes Associated Protein (YAP) downstream genes activation. This study specifically examines the effects of glutamine and serum deprivation on 23 YAP downstream genes that have been screened by previous laboratory research in HEp2 cancer cells using Immunostaining and qPCR. HEp2 cells were treated with two conditions of media, nutrient rich which have nutrients and nutrient deprived which gets deprived from glutamine and serum. Then, the cells get tested with CTPS antibodies for CTPS filament and harvested for the qPCR analysis. The results highlight the effect of nutrient deprivation through CTPS Immunostaining and nutrient deprivation effects towards YAP downstream gene with the downstream gene expression increase or decrease within the cell mRNA expression fold (2

-ΔΔCt). A total of 17 YAP downstream genes have been identified using t-test to be significantly increased or decreased (p<0.05), which all of it correlates with cellular function. All of the 17 influenced genes are correlated with cellular survival within nutrient deprivation conditions.