Experimental validation of PLAC8 in small intestinal neuroendocrine tumors

Abstract: Small intestinal neuroendocrine tumors (SI-NETs) are the most common malignant tumor in the small bowel. It is a small-growing cancer with good overall survival 5 years after diagnosis, but no full cure is available when in the metastatic stage. Placenta specific 8 (PLAC8) in chromosome 4 has been seen to have several roles in the cell-survival pathways, such as Nf-Kb/AKT/PI3K and autophagy regulations. Previous research has commented on PLAC8's potential role in gastronomic cancers, but it is unknown what function it has on SI-NETs. In this study, I measure PLAC8 expression in the cell line CNDT 2.5 with the use of RT-qPCR and western blot where I will compare RNA expressions and protein levels. I have transfected CNDT 2.5 cells with PLAC8 plasmids to create overexpression of the gene. Three assays were used to understand if PLAC8 overexpression changed the cell proliferation or cell-apoptotic functions of the cell; these were a proliferation assay, scratch wound assay,and apoptosis assay. The results showed an increased proliferation within PLAC8overexpressed cells compared to untreated cells. It did not have any significant change in the migration of the cells and PLAC8 overexpressed cells do not seem to have a change in its apoptotic events compared to pCMV6 transfected cells (empty vector). These results can help us understand what type of cellular pathways PLAC8 could influence, which in this study could be the Nf-Kb/AKT/PI3K or autophagy processes rather than the Epithelial-mesenchymal transition (EMT). These findings could in further studies help us find potential drugs which could target PLAC8 or its targets in the cell cycle.