Gene expression profiling and protein-protein network analysis revealed prognostic hub biomarkers linking cancer risk in type 2 diabetic patients.

Type 2 diabetes mellitus (T2DM) and cancer are highly prevalent diseases imposing major health burden globally. Several epidemiological studies indicate increased susceptibility to cancer in T2DM patients. However, genetic factors linking T2DM with cancer have been poorly studied. In this study, we followed computational approaches using the raw gene expression data of peripheral blood mononuclear cells of T2DM and cancer patients available in the gene expression omnibus (GEO) database. Our analysis identified shared differentially expressed genes (DEGs) in T2DM and three common cancer types, namely, pancreatic cancer (PC), liver cancer (LC), and breast cancer (BC). The functional and pathway enrichment analysis of identified common DEGs highlighted the involvement of critical biological pathways, including cell cycle events, immune system processes, cell morphogenesis, gene expression, and metabolism. We retrieved the protein-protein interaction network for the top DEGs to deduce molecular-level interactions. The network analysis found 7, 6, and 5 common hub genes in T2DM vs. PC, T2DM vs. LC, and T2DM vs. BC comparisons, respectively. Overall, our analysis identified important genetic markers potentially able to predict the chances of PC, LC, and BC onset in T2DM patients.

Yeast 2-hybrid assay for investigating the interaction between the centrosome proteins PLK4 and STIL.

Centrosomes act as a major microtubule-organizing center in many animal cells. This non-membranous cell organelle is involved in a wide range of cell functions like division, migration, and signaling. Centrosome houses hundreds of proteins that interact in a regulated manner to carry out their tasks. Most of these centrosome proteins have either the coiled-coil or disordered regions, making them challenging to purify for functional analysis. Yeast 2-hybrid (Y2H) is an in vivo based method for studying protein-protein interactions by exploiting the modular nature of yeast's GAL4 transcription factor. This method provides an alternative strategy for investigating bi-molecular interactions, especially when dealing with otherwise difficult proteins to purify by conventional biochemical approaches. We present a detailed protocol for showing the interaction between the two core centrosome proteins, i.e., PLK4 and STIL, using the Y2H assay.