Identification of Candidate Biomarkers and Cancer Genes AHNAK2 and EPPK1 in Pancreatic Cancer

Abstract

Aims: The lack of specific symptoms at early tumor stages, together with a high biological aggressiveness of the tumor contribute to the high mortality rate for pancreatic cancer (PC). Improved screening for earlier diagnosis, through the detection of diagnostic and prognostic biomarkers provides the best hope of increasing the rate of curatively resectable carcinomas. The aim of this study is to provide new targets for use as biomarkers in PC. Study Design: In a previous study, we identified novel candidate cancer genes and biomarkers that were significantly upregulated in PC, through a meta-analysis of large number of microarray datasets, using bioinformatics methods.  In this study, we analyzed the expression of these genes in a panel of pancreatic cancer cell lines by quantitative Reverse Transcription-PCR (qRT-PCR).  Place and Duration of Study: Department of Chemistry and Biochemistry and Department of Biology, University of Northern Iowa, USA, between June 2014 and Dec 2015. Methodology: We analyzed the expression of three genes, AHNAK2, EPPK1 and IGHG3 in a panel of seven standard PC cell lines, AsPC-1, BxPC-3, Capan-2, CFPAC-1, HPAF-II, PANC-1, and SW 1990 by Relative Quantification. qRT-PCR experiments were conducted in triplicate, and each experiment was replicated twice using different passages.  Results: AHNAK2 was significantly upregulated in all PC cell lines tested, with P values < 0.005 except for PANC-1 (P < 0.05). EPPK1 too was significantly upregulated (P < 0.05) in six of seven PC cell lines tested. While IGHG3 was nominally upregulated in all PC cell lines, upregulation was significant (P < 0.05) in only four PC cell lines. Conclusion: Our results confirm that AHNAK2 and EPPK1 are novel candidates for use as biomarkers in pancreatic cancer. IGHG3 does not appear to be a suitable candidate, due to its low levels of expression in both PC and control cell lines.