The effect of fructose on pancreatic cancer, pancreatic inflammation: İn vitro comperative study

OBJECTIVE: The aim of our study was to investigate the effects of fructose on pancreatic cancer. MATERIALS AND METHODS: The human pancreatic cancer cell line (PANC-1) was used in our study.In the cell culture study performed, the cells were planted in the flask using the appropriate medium (culture medium) under vitrocial conditions.The proliferation, passages and follow-up of the cells were examined regularly using Inverted Microscope.Fructose and glucose dissolved in low glucose DMEM were added to PANC-1 cells separately.Fructose and glucose-free cells were accepted as controls.The maximum proliferative effect values of the active substances were calculated in GraphPad Prism v5.0 Software.Combination experiments were also performed to see the effect of fructose and glucose together.The RNeasy Plus Kit (Qiagen) was used for the isolation of total RNA from the cells in which the combination of fructose, glucose and fructose + glucose, and the control cells containing fructose and glucose were not administered.CDNA synthesis was performed using the-All-in-OneTMFirst-Strand cDNA Synthesis Kit '(Genecopoeia) from appropriate isolated total RNAs.qPCR Array study was performed with "All-in-OneTM qPCR Mix" (Genecopoeia) to determine the expression levels of genes using cDNA samples.Primers designed specifically for our target genes (KRAS, NFKB, ERK2) were used.Real-time PCR data were analyzed by web-based RT2Profiller PCR Array Data Analysis version 3.5 (http://pcrdataanalysis.sabiosciences.com/pcr/arrayanalysis.php) using LightCycler 480 software (Roche) Cycle Threshold (Ct) values. RESULTS: At 72 hours, the value of fructose and glucose that increased the proliferation in the cell line was determined to be 20 mM for both substances.According to the isobologram analysis to evaluate the co-efficacy of both agents, the dose where fructose and glucose combination increased cell proliferation was found to be 15 mM.When HGK-DMEM was applied to the treated cells, it was determined that NFKB1 expression was 4.46, 3.29 and 6.52 times more in the cells which were applied fructose, glucose and fructose-glucose combination, respectively.The ratio of NFKB1 was found to be 4,65, 3,43 and 7,87 times, respectively, in the cells treated with fructose, glucose and fructose-glucose when compared to LGK-DMEM-treated cells.When HGK-DMEM was applied to the treated cells, it was found that NFKB1 expression was 11.47, 7.51 and 11.93 times, respectively. When cells treated with fructose , glucose and fructose-glucose combination compared to LGK-DMEM-treated cells CRAS expression was found to be 7.56, 4.95 and 7.87 times, respectively.Cells treated with fructose, glucose, and fructose-glucose combination compared to the HGK-DMEM cells were found to express 5,69, 1,4 and 4,45 times more ERK2 genes, respectively.When compared with HGK-DMEM cells the ratio of 4.5, 1.1 and 3.52 times more ERK2 gene was determined in the cells which were applied fructose, glucose and fructose-glucose combination, respectively. CONCLUSION: Our study showed that KRAS, ERK2 and NFKB1 gene expression increased in the pancreatic cancer cells more than glucose,shows synergistic effect on NFKB1 and KRAS expression when combined with fructose glucose.These genes encode proteins in key pathways in pancreatic cancer pathogenesis.The metabolism of fructose different from glucose in the metabolism of the metabolism will increase the expression of these genes and will be better understood by later studies.New studies should be conducted to elucidate its role in pathogenesis and to determine whether pathways in fructose metabolism can be used as a therapeutic target.