CX-5461 potentiates imatinib-induced apoptosis in K562 cells by stimulating KIF1B expression (Part 4)
Description
The selective RNA polymerase I inhibitor CX-5461 has been shown to be effective in treating some types of leukemic disorders. Emerging evidence suggests that combined treatments with CX-5461 and other chemotherapeutic agents may achieve enhanced effectiveness as compared to monotherapies. In the present study, we tested whether CX-5461 could potentiate the effect of imatinib in the human chronic myeloid leukemia cell line K562. Cells were divided into 3 treatment groups (with 3 independent biological replications in each group): control, imatinib (100 nM for 48 hr) alone, and imatinib+CX-5461 (100 nM for 48 hr). Genome-wide mRNA sequencing was performed.
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K562 cells were cultured in RPMI 1640 medium supplemented with 10% fetal bovine serum and antibiotics. Cells were divided into 3 treatment groups (with 3 independent biological replications in each group): control (Con), imatinib (IM) (100 nM for 48 hr) alone, and imatinib+CX-5461 (IM_CX) (100 nM for 48 hr). Total RNA was extracted using TRIZOL Reagent (Thermo Fisher, Waltham, MA, USA) following the manufacturer's instructions. RNA integrity was checked with an Agilent Bioanalyzer 2100 (Agilent Technologies, Santa Clara, CA, USA). Qualified total RNA was further purified by RNAClean XP Kit (Beckman Coulter, Brea, CA, USA) and RNase-Free DNase Set (QIAGEN, Germany). RNA concentration was determined using a NanoDrop ND-2000 spectrophotometer (Thermo Fisher). Library construction was performed using VAHTS Stranded mRNA-seq Library Prep Kit for Illumina (from Vazyme, Nanjing, Jiangsu Province, China). RNA sequencing was performed using a NextSeq Illumina550 platform (Illumina, San Diego, CA, USA) in a paired-end manner. RNA processing, library construction and sequencing services were provided by Shanghai Biotechnology Corporation (Shanghai, China). Differentially expressed genes were defined by the false discovery rate (q value) < 0.05 and fold change > 2.