INDUCTION OF HEPARIN-BINDING EPIDERMAL GROWTH FACTOR-LIKE GROWTH-FACTOR MESSENGER-RNA BY PROTEIN-KINASE-C ACTIVATORS
INDUCTION OF HEPARIN-BINDING EPIDERMAL GROWTH FACTOR-LIKE GROWTH-FACTOR MESSENGER-RNA BY PROTEIN-KINASE-C ACTIVATORS
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DOI：
10.1038/ki.1994.322
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发表时间：
1994-09-01
影响因子：
19.6
通讯作者：
TSAI, JH
中科院分区：
文献类型：
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作者：
TAN, MS;TSAI, JC;TSAI, JH
肝素结合表皮生长因子样生长因子（HB-EGF）是一种强效的巨噬细胞来源的平滑肌细胞有丝分裂原。为了确定HB-EGF基因是否在系膜细胞中转录和调节，我们通过RNA印迹分析测定了培养的大鼠系膜细胞中HB-EGF mRNA的水平。在未受刺激的系膜细胞中检测到2.5kb的HB-EGF mRNA。蛋白激酶C激活剂12-O-十四烷酰佛波醇13-乙酸酯（TPA）可使系膜细胞中HB-EGF mRNA水平增加15倍，TPA对HB-EGF mRNA的诱导具有时间和剂量依赖性。10%胎牛血清、离子霉素、凝血酶和内皮素-1也能刺激HB-EGF mRNA的表达。蛋白激酶C抑制剂Staurosporine可阻断TPA和血清对HB-EGF mRNA的诱导作用。为了确定HB-EGF是否对系膜细胞具有促有丝分裂作用，我们用HB-EGF表达质粒转染COS细胞。转染这些质粒的COS细胞培养液以剂量依赖性方式增加系膜细胞中H-3-胸苷的掺入。据我们所知，这是第一个报告，HB-EGF在肾细胞中表达。HB-EGF的这种可诱导的转录表明它可能在肾脏疾病的系膜细胞增殖中具有自分泌作用。
Heparin-binding epidermal growth factor-like growth factor (HB-EGF) is a potent smooth muscle cell mitogen of macrophage origin. To determine whether the HB-EGF gene is transcribed and regulated in mesangial cells, we measured HB-EGF mRNA levels in cultured rat mesangial cells by RNA blot analysis. A 2.5-kb HB-EGF mRNA was detected in unstimulated mesangial cells. The protein kinase C activator 12-O-tetradecanoylphorbol 13-acetate (TPA) increased HB-EGF mRNA levels by 15-fold in mesangial cells, and this induction of HB-EGF mRNA by TPA was both time- and dose-dependent. HB-EGF mRNA could also be stimulated by 10% fetal calf serum, ionomycin, thrombin, and endothelin-1. Staurosporine, a protein kinase C inhibitor, abolished the induction of HB-EGF mRNA by TPA and serum. To determine whether HB-EGF is mitogenic for mesangial cells, we transfected COS cells with HB-EGF expression plasmids. Culture medium from COS cells transfected with these plasmids increased H-3-thymidine incorporation in mesangial cells in a dose-dependent manner. To our knowledge, this is the first report that HB-EGF is expressed in renal cells. This inducible transcription of HB-EGF suggests that it may have an autocrine role in mesangial cell proliferation in kidney disease.