Time point studies of bPGN treated HCT-116 cell line

HCT-116 cell lines were treated with bPGN and treated cells were harvested at time-point 0, 1hour and 24hours. Identification of RNA-interacting pharmacophores could provide new chemical probes and potentially novel RNA-based therapeutics. Herein, using a high-throughput differential scanning fluorimetry assay, we identified small molecule natural products with the capacity to bind the discrete stem-looped structure of pre-miR-21. The most potent compound identified was a prodiginine-type compound, butylcylcoheptyl prodiginine (bPGN), with the ability to inhibit Dicer-mediated processing of pre-miRNA-21 in vitro and in cells. Time dependent RT-qPCR, western blot, and transcriptomic analyses showed modulation of miR-21 expression and its target genes such as PDCD4 and PTEN upon treatment with bPGN, supporting on-target inhibition. Consequently, inhibition of cellular proliferation in HCT-116 colorectal cancer cells was also observed when treated with bPGN. The discovery that bPGN can bind and modulate the expression of regulatory RNAs such as miR-21 helps set the stage for further development of this class of natural product as a molecular probe or therapeutic agents against miRNA-dependent diseases. Cells were treated with bPGN and cells were harvested for nanostring analysis at time-points 0, 1, and 24 hours. Contributor: NCI Genomics Center