Ccl 2

Four primers, CCL2, CXCL8, CXCL10, and PTGS2, were purchased from Sangon Biotech Corporation (Shanghai, China). The sequence details are listed in Table S1. HCT116 cells were intervened with the above 3 drugs at a concentration of 20 µM for 48 hours. After that, the cells were collected, and RNA was extracted using the TRIzol method. After the purity and integrity of the RNA had been determined, cDNA was prepared through reverse transcription. The PCR conditions were the following: pre-denaturation at 95 °C for 1 minute; denaturation at 95 °C for 15 seconds, annealing at 58 °C for 20 seconds, extension at 72 °C for 20 seconds, for 40 cycles; then, extension at 72 °C for 5 minutes to terminate the reaction. After the completion of real-time quantitative PCR (RT-qPCR), the reliability of the melting curve and amplification curve results obtained by PCR was quantitatively analyzed, and the cycle threshold (Ct) was set. The ratios of CCL2, CXCL8, CXCL10, PTGS2 and the internal control, β-actin, were used to represent their relative expression levels, and the relative mRNA expression intensity of each was calculated. There were three duplicate holes in each group, and the test was repeated 3 times.