| Abstract [eng] |
Cancer is a disease marked by uncontrolled proliferation of cells, which in spite of great efforts remains one of the most urgent public health issues and is predicted to increase in occurrence. Colorectal cancer in particular was one of the most frequently reported forms of cancer in 2022, making up 9.6% of cases. Many of the currently approved chemotherapeutic agents exhibit adverse effects. Consequently, chemotherapy can be greatly bolstered by the discovery of more effective and safe chemotherapeutic agents. This work aimed to investigate the effects of a set of triazole-based compounds on the pre-mRNA splicing of tumour-associated genes in HCT116 cells, including the evaluation of the effects of the triazole-based compounds on the viability of cells. Also, to compare tumour-associated mRNA isoform formation under normoxia and hypoxia, and to evaluate the effects of selected triazole-based compounds on the mRNA isoform formation under the same conditions. The HCT116 cells in normoxia or hypoxia (72h) were cultivated. The effects on cell viability were assessed using the MTT assay. The half-maximal inhibitory concentrations (IC50) of selected compounds was determined. mRNAs of selected genes were isolated, complimentary DNA was synthesised and then amplified by PCR. These results were visualised via electrophoresis and examined using bioinformatical analysis tools. Assessment of the effects on HCT116 cell viability of 17 triazole-based compounds, revealed that compounds IT123, IT126 and IT135 were the most effective, reducing cell viability to 8-22%. Their IC50 values were determined to be 51.39 μM, 41.23 μM and 53.09 μM, respectively. Analysis of FAS, FGFR1OP, PUF60, APAF1, CDC42BPA and CASP9 pre-mRNA splicing in cells cultured under either normoxia or hypoxia, indicated that hypoxia influences mRNA isoform formation. Assessment of the effects of selected compounds on pre-mRNA splicing in cells cultured either under normoxia or hypoxia, showed that these compounds have distinct effects on pre-mRNA splicing of different genes. In normoxia FAS pre-mRNA splicing was altered by all compounds, while under hypoxia only by compounds IT126 and IT135. PUF60 and APAF1 pre-mRNA splicing was affected only by IT123 under normoxia, while CDC42BPA mRNA isoform formation was only affected by IT126 and IT135 under normoxia. All three compounds had no effect on FGFR1OP and CASP9 isoform formation. |
