The anticancer Ruthenium drug BOLD-100 regulates apoptosis of BRAFMT colorectal cancer cells through modulation of the AhR/ROS/ATR signalling axis.

The understanding of how oncogenes modulate the mechansim of action of compounds relies on interrogating acute and longer transcriptional changes following treatment. We report here the transcriptional profiles of an isogenic cell line model VACO432 (BRAFMT V600E/WT) and VT1 (BRAFWT/-) following 3H and 24H treatment with a novel ruthenium compound BOLD-100. We compare the two cell line models based on their BRAF status to identify transcriptional changes modulted by oncogenic BRAF. Overall design: Bulk RNA-sequencing was performed for VACO432 and VT1 cells treated with IC30 dose (24µM) BOLD-100 for 3h and 24h. Three biological replicates for each sample were included in the experiment. Total RNA was extracted as described in the main materials and methods section. Each sample underwent QC, and library preparation was performed with 100ng RNA (Illumina NextSeq 500 kit, Illumina, UK) according to the manufacturer's instructions. using the KAPA RNA HyperPrep Kit with RiboErase (HMR) (Roche, USA). Paired 91 end sequencing was conducted with a sequencing depth of 50 million reads, resulting in 75-base pair reads. The reads were trimmed based on Phred score from the 3-prime end and then aligned to the human genome (hg38) using TopHat21 (version 2.1.0) with default settings.