Zhang Chunyan, Chang Cuifang, Zhao Weiming, Gao Hang, Wang Qiwen, Li Deming, Zhang Fuchun, Zhang Shifu, Xu Cunshuan
Xinjiang Key Laboratory of Biological Resources and Genetic Engineering, College of Life Science and Technology, Xinjiang University, Urumqi, China.
State Key Laboratory Cultivation Base for Cell Differentiation Regulation, College of Life Science, Henan Normal University, Xinxiang, Henan, China.
PLoS One. 2017 Jul 27;12(7):e0180607. doi: 10.1371/journal.pone.0180607. eCollection 2017.
Our previous study has proved that the chromosome 9 open reading frame 116 (C9orf116) (NM_001106564.1) was significantly up-regulated in the proliferation phase of liver regeneration. To study its possible physiological function, we analyzed the effect of C9orf116 on BRL-3A cells via over-expression and interference technique. MTT results showed that the cell viability of the interference group was significantly lower than the control group at 48h after transfection (P<0.05), whereas it was significantly higher in the over-expression group (P<0.05). The flow cytometry results showed that C9orf116 knockdown or over-expression had little effect on BRL-3A cell apoptosis. However, the number of cells in division phase (G2/M) was significantly reduced in the interference group (P<0.05), but significantly increased in the over-expression group (P<0.01). Furthermore, the expressions of cell proliferation-related genes CCNA2, CCND1 and MYC both at mRNA and protein levels were down-regulated in the interference group and up-regulated in the over-expression group. Therefore, we concluded that C9orf116 may promote cell proliferation by modulating cell cycle transition and the expression of key genes CCNA2, CCND1 and MYC in BRL-3A cells.
我们之前的研究已证明，9号染色体开放阅读框116（C9orf116）（NM_001106564.1）在肝再生的增殖期显著上调。为研究其可能的生理功能，我们通过过表达和干扰技术分析了C9orf116对BRL-3A细胞的影响。MTT结果显示，转染后48小时，干扰组的细胞活力显著低于对照组（P<0.05），而过表达组的细胞活力则显著更高（P<0.05）。流式细胞术结果显示，C9orf116的敲低或过表达对BRL-3A细胞凋亡影响不大。然而，干扰组中分裂期（G2/M）的细胞数量显著减少（P<0.05），而过表达组中则显著增加（P<0.01）。此外，干扰组中细胞增殖相关基因CCNA2、CCND1和MYC在mRNA和蛋白质水平的表达均下调，而过表达组中则上调。因此，我们得出结论，C9orf116可能通过调节细胞周期转换以及BRL-3A细胞中关键基因CCNA2、CCND1和MYC的表达来促进细胞增殖。
