a Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences , Moscow , Russian Federation.
b Centre for Investigative and Diagnostic Oncology, Department of Natural Sciences, School of Science and Technology , Middlesex University , London , United Kingdom.
Cell Cycle. 2017 Oct 18;16(20):1979-1991. doi: 10.1080/15384101.2017.1371880. Epub 2017 Sep 5.
The nucleolar proteins which link cell proliferation to ribosome biogenesis are regarded to be potentially oncogenic. Here, in order to examine the involvement of an evolutionary conserved nucleolar protein SURF6/Rrp14 in proliferation and ribosome biogenesis in mammalian cells, we established stably transfected mouse NIH/3T3 fibroblasts capable of conditional overexpression of the protein. Cell proliferation was monitored in real-time, and various cell cycle parameters were quantified based on flow cytometry, Br-dU-labeling and conventional microscopy data. We show that overexpression of SURF6 accelerates cell proliferation and promotes transition through all cell cycle phases. The most prominent SURF6 pro-proliferative effects include a significant reduction of the population doubling time, from 19.8 ± 0.7 to 16.2 ± 0.5 hours (t-test, p < 0.001), and of the length of cell division cycle, from 17.6 ± 0.6 to 14.0 ± 0.4 hours (t-test, p < 0.001). The later was due to the shortening of all cell cycle phases but the length of G1 period was reduced most, from 5.7 ± 0.4 to 3.8 ± 0.3 hours, or by ∼30%, (t-test, p < 0.05). By Northern blots and qRT-PCR, we further showed that the acceleration of cell proliferation was concomitant with an accumulation of rRNA species along both ribosomal subunit maturation pathways. It is evident, therefore, that like the yeast homologue Rrp14, mammalian SURF6 is involved in various steps of rRNA processing during ribosome biogenesis. We concluded that SURF6 is a novel positive regulator of proliferation and G1/S transition in mammals, implicating that SURF6 is a potential oncogenic protein, which can be further studied as a putative target in anti-cancer therapy.
与核糖体生物发生相关的核仁蛋白被认为具有潜在的致癌性。在这里,为了研究进化保守的核仁蛋白 SURF6/Rrp14 参与哺乳动物细胞增殖和核糖体生物发生的情况,我们建立了能够条件性过表达该蛋白的稳定转染的小鼠 NIH/3T3 成纤维细胞系。实时监测细胞增殖,并基于流式细胞术、Br-dU 标记和常规显微镜数据定量各种细胞周期参数。我们发现 SURF6 的过表达加速了细胞增殖并促进了细胞周期的所有阶段的过渡。SURF6 最显著的促增殖作用包括群体倍增时间从 19.8 ± 0.7 小时显著缩短至 16.2 ± 0.5 小时(t 检验,p < 0.001),细胞分裂周期长度从 17.6 ± 0.6 小时显著缩短至 14.0 ± 0.4 小时(t 检验,p < 0.001)。后者是由于所有细胞周期阶段的缩短,但 G1 期的长度缩短最多,从 5.7 ± 0.4 小时缩短至 3.8 ± 0.3 小时,或缩短约 30%(t 检验,p < 0.05)。通过 Northern 印迹和 qRT-PCR,我们进一步表明细胞增殖的加速伴随着核糖体亚基成熟途径中 rRNA 物种的积累。因此,与酵母同源物 Rrp14 一样,哺乳动物 SURF6 参与核糖体生物发生过程中 rRNA 的各种加工步骤。我们得出结论,SURF6 是哺乳动物增殖和 G1/S 过渡的新型正调控因子,这表明 SURF6 是一种潜在的致癌蛋白,可以进一步作为抗癌治疗的潜在靶点进行研究。