Evaluation of Anti-proliferative Activity of Quercetin on Pancreas Cancer Cells and its effect on important microRNAs

Abstract

Pancreatic cancer is one of the most lethal cancers worldwide. The resistant nature of this cancer increases the need for newer compounds to be combined with conventional chemotherapy. It appears that Quercetin is a flavonoid with excellent properties for this purpose. MicroRNAs are small, non-coding RNA molecules that regulate gene expression. They can be used as biomarkers, prognostic factors, or therapeutic targets for some cancers. Quercetin may improve the effect of chemotherapy on microRNA profile in pancreatic cancer in-vitro.Aims: This study aimed to investigate the effect of the combination of Quercetin with 5-FU on the expression level of selected microRNAs in vitro.Methods: The cytotoxicity of Quercetin on pancreatic cancer cell line was investigated using the MTT assay. Next, total RNA was extracted from cancer cells in four groups (control, treatment with Quercetin, treatment with 5-FU, and treatment with a combination of Quercetin + 5-FU). Then, specific cDNA for miR-21, miR-133, miR- 155, miR-181 and miR-451 was prepared. For this purpose, specific RT primers was designed for each of the mentioned microRNAs separately. Finally, the target microRNAs expression levels were checked using the Real-Time PCR method.Results: IC50 of Quercetin was determined 50 µM. The real-time PCR result is as follows: The expression levels of miR-21 and miR-155 were significantly reduced in all groups compared to the control group. The expression of miR-133 was significantly increased in the combination group. miR-181 did not show meaningful changes. miR-451 was up-regulated in all groups. The c-Myc gene was chosen as a common target of these microRNAs. Both real-time PCR and Western Blot tests showed a significant reduction in all groups compared to the control group.Conclusion: Based on the results, Quercetin can potentiate 5-FU's anti-proliferative effect in the Mia-PaCa-2 cell line.