The effect of combination of resveratrol and BIBR1532 on the LncRNA expression in colorectal cancer cells

Colorectal cancer is known as the third most common cancer type and the fourth most common cause of death and thus new targeted therapies are continuously investigated. In cancer cells; it is known that certain types of long non-coding RNAs are highly expressed, and these lncRNAs are involved in development and progression of carcinomas. Therefore, lncRNAs are thought as a target for cancer therapies. BIBR1532 is a telomerase inhibitor and in cancer research, it is used to inhibit active telomerase in tumours. Resveratrol is a natural phytoalexin and it targets the molecules that are involved in proliferation and apoptosis. In addition, resveratrol induces apoptosis by inhibiting KRAS-mediated signalling. In this study, the aim is to apply BIBR1532 and resveratrol agents to HT-29 colorectal cancer cells and to determine cyototoxicity of these agents. Following this, possible apoptotic effects of BIBR1532 and resveratrol will be investigated. Finally, after exposure to these two agents, lncRNA expression changes will be determined in HT-29 cells. The concentration of HT-29 cells in each well of a 96-well plate was determined with the proliferation experiment. With WST-1 analysis cytotoxic effects of BIBR1532 and resveratrol were investigated depending on time and dose. After this, pre-determined IC50 doses were applied to HT-29 cells both single and in combination and the cells with no drug application, were taken as control.Then apoptosis test was performed via Annexin V assay. For lncRNA analysis, agents were applied as in the apoptosis test and again the cells with no drug application, were taken as control. RNA isolation from all cell groups was carried out and cDNA was synthesized, then RT-PCR was performed for the examination of changes in lncRNA levels. For HT-29 cells, optimal cell concentration was determined as 2x104 cells/ ml and according to cytotoxicity assay IC50 doses were decided for BIBR1532 and resveratrol to be 50.81 μM (48. hour) and 86.23 μM (72 hour) respectively. CI was 1.7 for these two agents. Viability of cells after exposure to both BIBR1532 and resveratrol agents, decreased; however there was no apoptosis inducing effect found for both agents. There is no study about the application of BIBR1532 and resveratrol combination on CRC cells in literature. We suggest that the investigation of the effects of these agents on cancer related lncRNAs and linking lncRNAs with CRC will give new point of view in generating new treatment strategies.