Fig. 2
Silencing NAT1 enhances the chemo-resistance of chemotherapeutic agents in CRC cells. A Using the oncoPredict package, we calculated the relative IC50 values for the top five chemotherapeutic agents in each sample based on RNA-seq data from TCGA-CRC (n = 592). Spearman correlation analysis was conducted to evaluate the relationship between NAT1 expression and the relative IC50 values of these top five chemotherapeutic agents. B The expression levels of NAT1 in each group were assessed using Western blot (WB) assays. C The IC50 values (uM) for each group were determined using the CCK-8 assay. D Flow cytometry was employed to quantify the mean fluorescence intensity of annexin-V and caspase-3 in each group. E Cells were exposed to DMSO or 5 ng of each compound—equivalent to 0.55 μM vinblastine, 0.62 μM docetaxel, 1.90 μM gemcitabine, 0.54 μM vincristine, and 1.28 μM daporinad—for an initial 24-h treatment period. This was followed by a 24-h recovery phase in fresh complete medium. The cycle of 24 h of drug exposure followed by 24 h of recovery in fresh medium was repeated for a total of five cycles. WB analysis revealed that this treatment regimen led to a downregulation of NAT1 expression in CRC cells. Statistical significance is denoted as ns represents no significance, *P < 0.05. μM represents micromolar. Comparison of sh-NC with sh-NAT1 and OE-NC with OE-NAT1 via unpaired t-test. The experiments were independently repeated at least three times