Altered gene expression in CHO cells following polyamine starvation.
AIM
To investigate the impact of polyamine deprivation on the transcriptome of CHO cells RESULTS: Polyamines play a central but poorly-understood role in cell proliferation. Most studies to date have utilised chemical inhibitors to probe polyamine function. Here we exploit the fact that CHO cells grown in serum-free medium have an absolute requirement for putrescine supplementation due to their deficiency in activity of the enzyme arginase. A gene expression microarray (Affymetrix) analysis of CHO-K1 cells starved of polyamines for 3 days showed that cessation of growth, associated with increased G1/S transition and inhibition of M/G1 transition was accompanied by increased mRNA levels of mitotic complex checkpoint genes (Mad2l1, Tkk, Bub1b) and in the transition of G1- to S-phase (such as Skp2 and Tfdp1). mRNAs associated with DNA homologous recombination and repair (including Fanconi's anaemia-related genes) and with RNA splicing were consistently increased. Alterations in mRNA levels for genes related to protein processing in the ER, to ER stress, and to p53-related and apoptosis pathways were also observed. mRNAs showing highest levels of fold-change included several which code for membrane-localised proteins and receptors (Thbs1, Tfrc1, Ackr3, Extl1).


CONCLUSIONS
Growth-arrest induced by polyamine deprivation was associated with significant alterations in levels of mRNAs associated with cell cycle progression, DNA repair, RNA splicing, ER trafficking and membrane signalling as well as p53 and apoptosis-related pathways.
['Capella Roca|Berta|B|http://orcid.org/0000-0001-7071-2677', 'Doolan|Padraig|P|', 'Barron|Niall|N|', "O'Neill|Fiona|F|", 'Clynes|Martin|M|']
D000818:Animals D016466:CHO Cells D002470:Cell Survival D006224:Cricetinae D003412:Cricetulus D003470:Culture Media D005786:Gene Expression Regulation D011700:Putrescine D059467:Transcriptome
Gene expression Polyamines Transcriptomics Putrescine Chinese hamster ovary cells (CHO)