Abstract 1229 ''Targeting Notch, IL-1 and Leptin crosstalk may have promising approach in HCT-15 colorectal cancer cells''

Objective: The interaction between NILCO signaling and VEGF is associated with metastasis and decreased survival rate. Previous studies have shown that leptin plays a pivotal role in the crosstalk between NILCO and angiogenic VEGF. Additionally, it has been shown that matrix metalloproteinases (MMPs) are induced by leptin, causing tumor cells to spread from the basement membrane and contributing to tumor angiogenesis. In this study, we aimed to determine the effect of targeting NILCO in human colorectal cancer (CRC). Methods: HCT-115 cells were transfected with Notch, IL-1, and leptin small interfering RNAs (siRNA) using lipofectamine 2000 reagent according to the manufacturers' instructions. Notch1, Cas-9 and VEGFA mRNA gene levels were determined by qRT-PCR; leptin, VEGF-A and TIPM-1 protein levels were determined by WB, and leptin, VEGF-A and IL-1β protein levels were determined by ELISA. Statistical significance was tested with with one-way ANOVA and post hoc Tukey tests using GraphPad Prism6. Results: Inhibition of leptin by siRNA decreased Notch mRNA levels and, leptin and VEGFA protein levels.According to ELISA results; leptin knockdown decreased leptin, IL-1 and VEGFA protein levels. Inhibition of Notch1 by siRNA decreased Notch1 mRNA levels, leptin protein levels and, increased TIMP-1 protein level. According to ELISA results, downregulation of Notch1 decreased leptin, IL-1 and VEGF-A protein levels. IL-1 siRNA increased cas-9 mRNA levels and, decreased VEGF-A mRNA and VEGFA protein levels. According to ELISA results, IL-1 siRNA decreased leptin, IL-1 and VEGFA protein levels. Conclusion: In conclusion, our study demonstrates that communication between members of the NILCO complex triggers pro-angiogenic and invasive signals that induce colorectal tumor development. Targeting of the NILCO complex may be a promising approach to treatment of CRC. The present study was supported by Eskisehir Osmangazi University Scientific Research Projects (BAP) Coordination Unit (grant no. 202011C102).