Requirement of cyclin D1 in mesangial cell mitogenesis
Requirement of cyclin D1 in mesangial cell mitogenesis
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DOI：
10.1681/asn.v1181398
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发表时间：
2000-08-01
影响因子：
13.6
通讯作者：
Schöcklmann, HO
中科院分区：
文献类型：
--
作者：
Lang, S;Hartner, A;Schöcklmann, HO
系膜细胞增生是肾小球肾炎的常见表现。研究了细胞周期蛋白D1（cyclin D1）对MC增殖的调控作用。在系膜增生性肾小球肾炎大鼠模型中，在疾病的第5天，MC增生的高峰之前，细胞周期蛋白D1阳性MC核的数量增加是突出的。在生长停滞的大鼠MC培养中，促有丝分裂刺激血清或血小板衍生生长因子（PDGF）导致细胞周期蛋白D1蛋白表达迅速增加。转化生长因子-β 1在12 h时抑制PDGF对细胞周期蛋白D1蛋白的诱导。在检查细胞周期蛋白D1的亚细胞分布。观察到用PDGF刺激MC 6小时引起细胞周期蛋白D1从细胞质易位到细胞核中。与PDGF和转化生长因子-β 1共孵育完全抑制了这种作用，而不改变该时间点的细胞周期蛋白D1蛋白丰度。为了测试细胞周期蛋白D1蛋白水平的降低是否足以抑制有丝分裂，用与大鼠细胞周期蛋白D1 mRNA互补的反义寡核苷酸（ODN）转染MC。针对细胞周期蛋白D1的反义ODN使血清或PDGF诱导的细胞周期蛋白D1的蛋白表达分别降低至对照水平的27%或10%。这些抑制作用与细胞周期蛋白依赖性激酶4活性降低相关。cyclin D1反义ODN也能降低PDGF诱导的p21（Waf-1）蛋白水平的升高。用[H-3]胸腺嘧啶核苷摄取和细胞计数法测定cyclin D1反义ODN对血清或PDGF诱导的MC增殖有明显抑制作用。结论：MC增殖需要细胞周期蛋白D1蛋白表达增加。靶向细胞周期蛋白D1的表达可能是一种有效的手段，在体外和体内抑制MC增殖。
Hyperplasia of mesangial cells (MC) is a frequent finding in glomerulonephritis. The control and function of cyclin D1, a regulator of cell cycle progression, in MC proliferation in vivo and in vitro were investigated. In a rat model of mesangioproliferative glomerulonephritis, increases in the number of cyclin D1-positive MC nuclei were prominent on day 5 of the disease, preceding the peak of MC hyperplasia. In growth-arrested rat MC in culture, mitogenic stimulation with serum or platelet-derived growth factor (PDGF) led to rapid increases in cyclin D1 protein expression. Transforming growth factor-beta 1 inhibited PDGF induction of cyclin D1 protein at 12 h. In an examination of the subcellular distribution of cyclin D1. it was observed that stimulation of MC with PDGF for 6 h caused translocation of cyclin D1 from the cytoplasm into the nucleus. Coincubation with PDGF and transforming growth factor-beta 1 completely inhibited this effect, without altering the cellular cyclin D1 protein abundance at that time point. To test whether reduction of cyclin D1 protein levels was sufficient to inhibit mitogenesis, MC were transfected with antisense oligonucleotides (ODN) complementary to rat cyclin D1 mRNA. Antisense ODN against cyclin D1 reduced the serum- or PDGF-induced protein expression of cyclin D1 to 27 or 10% of control levels, respectively. These inhibitory effects were correlated with diminished cyclin-dependent kinase 4 activity. Antisense ODN against cyclin D1 also decreased the PDGF-induced increase in p21(Waf-1) protein levels. The MC proliferation caused by serum or PDGF was markedly inhibited by antisense ODN against cyclin D1, as measured by [H-3]thymidine uptake and cell counts. It is concluded that increased cyclin D1 protein expression of MC is required for MC proliferation. Targeting cyclin D1 expression may represent an effective means to inhibit MC proliferation in vitro and in vivo.