Effect of rotary cell culture system-simulated microgravity environment on the expression of lncRNA in mouse fibroblasts / 解放军医学杂志

ABSTRACT

Objective To investigate the effects of simulated microgravity by rotary cell culture system (RCCS) on expression profiles of long non-coding RNA (lncRNA) in mouse fibroblasts L929 cell line.Methods L929 cells were cultured in vitro and randomly divided into simulated microgravity (SMG) group and normal gravity (NG) group.Each group had three samples,the rotator axis of SMG group was paralleled to the ground rotation,while the rotator axis of NG group was vertical to the ground rotation,and the speed of rotation was consistent for the two groups.The samples of two groups were collected on 7th day of culture and the total RNAs were extracted,labeled and hybridized in sequence.The lncRNA and mRNA were detected by Agilent Mouse lncRNA Chips respectively.Differentially expressed lncRNA were identified and then validated by RT-qPCR.GO and Pathway analysis were applied to determine the functional distribution of these target genes.The integration predictions of the lncRNA and mRNA co-expression had been proposed to refine the functional lncRNA-mRNA relationships.Results There were 238 differentially expressed lncRNAs including 134 lncRNAs up-regulated and 104 lncRNAs down-regulated,and 237 differentially expressed mRNAs including 53 mRNAs up-regulated and 184 mRNAs down-regulated significantly in mouse fibroblasts L929 cell line under simulated microgravity by RCCS.The RT-qPCR showed a high concordance with chip microarray results in 4 differentially expressed lncRNA.GO analysis showed that the differentially expressed lncRNAs were related to the biological processes such as negative regulation of megakaryocyte differentiation and negative regulation of wound healing.Pathway analysis showed that these target genes were related to the signal pathways of systemic lupus erythematosus and TGF-β.The lncRNA-mRNA co-expression networks were also established.Conclusion The simulated microgravity by RCCS could significantly affect the expression profiles of lncRNA and mRNA in mouse fibroblasts L929.The lncRNA target genes prediction and functional enrichment analysis based on gene chip technology may provide the theoretical basis for illustrating the mechanism and management of weightlessness stress injury.