The combination ofSPP1knockdown and gemcitabine treatment enhances apoptosis and reduces invasiveness of pancreatic cancer cells

Abstract

Pancreatic ductal adenocarcinoma (PDAC) is poised to be a leading cause of cancer-related deaths. Despite developing new treatment strategies, patient outcomes have not significantly improved. Chemoresistance has been implicated as a major contributor to ineffective treatments observed with studies suggesting combination therapy targeting multiple pathways. This study explored dysregulated genes in tumours of PDAC patients to identify targets which could be used effectively in combination with conventional therapy against cancer cells. In this study, PCR arrays were used for gene expression profiling of tumours obtained from South African PDAC patients to identify key differentially expressed pathways and potentially new therapeutic target genes. SPP1 was selected and RNA interference was used to knock the gene down. Migration and apoptosis assays were used to evaluate the effect of the knockdown, alone and in combination with Gemcitabine, on a pancreatic cancer cell line, MIA PaCa-2. Proteomic analysis using SWATH-MS was used to demonstrate potential molecular mechanisms linked to the morphological and phenotypical effects observed with treatment. We demonstrated several genes linked to the growth factor and signal transduction signalling pathways, and identified SPP1 as a target. We observed that by combining SPP1 knockdown with conventional chemotherapy, gemcitabine, resulted in a synergistic effect, leading to an enhanced early apoptotic response. A decline of migratory and invasive capabilities of MIA PaCa-2 cells was observed upon subjecting the cancer cells to SPP1 reduction and gemcitabine treatment. Furthermore, proteomic analyses uncovered several pathways that were dysregulated by the combination therapy including both pro-and anti-tumorigenic ones. The study findings indicate that SPP1 could be a potential therapeutic target for PDAC, and the possible synergistic effects observed when SPP1 knockdown was combined with gemcitabine treatment suggest a potential avenue for developing more effective treatments for PDAC while exploring tumour cell adaptation for survival.