Project description:We employed high-throughput sequencing to profile the non-coding RNA content of primary ESC and Fb derived exosomes. All 6 libraries were sequenced to a comparable depth (ESC1: 12.3M, ESC2: 14M, ESC3: 12.8M, FB1: 14.7M, FB2: 13.7M, FB3: 12.3M) and exhibited a very low proportion of adapter-only reads or fragments shorter than 17bp (4.4 ± 0.5% ), showing efficient fragment incorporation during library preparation. All libraries had high small RNA content (>50% successfully aligned and assigned to small RNAs). ESC libraries had significantly higher small RNA content (ESC: 84.5 ± 0.005% vs FB: 60.5 ± 0.05%, p<0.05, two-sided Welch’s t-test). ESC samples show remarkably higher proportions of miRNAs (~9-fold higher, p=3.6*10-5 ) and snRNAs (~12-fold higher, p=0.002) compared to Fb. Fibroblasts exhibited higher RPM of reads mapping on lncRNAs (4-fold, p=0.052)
Project description:Microarray analysis was carried out on human MOH cells exposed to fb-PMT (10e-6 M tetrac equivalent) for 24 h. fb-PMT significantly downregulated electron transport chain genes ATP5A1 (ATP synthase A1), ATP51, ATP5G2, COX6B1 (cytochrome c oxidase subunit 6B1), NDUFA8 (NADH dehydrogenase [ubiquinone] FA8), NDUFV2 and other NDUF genes, SLC25A6 (solute carrier group 2), UCP2 ([mitochondrial] uncoupling protein 2) and COX5A. The NDUF and ATP genes are also relevant to control of oxidative phosphorylation and transcription. Six additional NDUF genes linked to oxidative phosphorylation were affected. Conclusions. fb-PMT caused downregulation of expression of a panel of genes involved in electron transport, oxidative phosphorylation, and cytoplasmic ribosomal protein generation in MOH cells. A key component of the anticancer activity of fb-PMT relates to disordering of ATP generation mechanisms in tumor cells.