Clemmons D R, Van Wyk J J
J Clin Invest. 1985 Jun;75(6):1914-8. doi: 10.1172/JCI111906.
Cultured porcine aortic smooth muscle cells and human fibroblasts produce somatomedinlike peptides and secrete them into the surrounding microenvironment. This production has been linked to their ability to replicate. The objective of this study was to determine if a specific anti-somatomedin-C (Sm-C) monoclonal antibody that binds the somatomedinlike peptides could inhibit replication by porcine aortic smooth muscle cells and human fibroblasts. To determine if the antibody could inhibit the effect of endogenously produced somatomedinlike peptide, increasing concentrations of antibody were co-incubated with platelet-derived growth factor, a known stimulant of somatomedinlike peptide secretion, and Sm-C-deficient platelet-poor plasma. Addition of the antibody reduced fibroblast [3H]thymidine incorporation from 35,100 +/- 500 to 10,600 +/- 700 cpm (P less than 0.001), and in smooth muscle cells from 29,600 +/- 1,800 to 10,800 +/- 1,100 cpm (P less than 0.001). Co-incubation of exogenously added Sm-C (20 ng/ml) with maximally inhibitory dilutions of antibody increased [3H]thymidine incorporation in fibroblasts from 7,800 +/- 1,000 to 18,900 +/- 800 cpm (P less than 0.01), and in smooth muscle cells from 9,800 +/- 1,200 to 17,200 +/- 1,100 cpm (P less than 0.01). Insulin, which can substitute for Sm-C as a mitogen and does not bind to the antibody, stimulated DNA synthesis when co-incubated with the antibody, thereby excluding the possibility of nonspecific cytotoxicity. These results strengthen the hypothesis that the rate of DNA synthesis of these two cell types in vitro is directly linked to their capacity to produce somatomedinlike peptides. They further support the cellular production of somatomedinlike peptides as examples of the autocrine model of growth regulation.
培养的猪主动脉平滑肌细胞和人成纤维细胞可产生类生长调节素肽,并将其分泌到周围的微环境中。这种产生与它们的复制能力有关。本研究的目的是确定一种能结合类生长调节素肽的特异性抗生长调节素C(Sm-C)单克隆抗体是否能抑制猪主动脉平滑肌细胞和人成纤维细胞的复制。为了确定该抗体是否能抑制内源性产生的类生长调节素肽的作用,将浓度不断增加的抗体与血小板衍生生长因子(一种已知的类生长调节素肽分泌刺激物)和缺乏Sm-C的少血小板血浆共同孵育。加入该抗体后,成纤维细胞的[³H]胸腺嘧啶核苷掺入量从35100±500 cpm降至至10600±700 cpm(P<0.001),平滑肌细胞的掺入量从29600±1800 cpm降至10800±1100 cpm(P<0.001)。将外源性添加的Sm-C(20 ng/ml)与最大抑制稀释度的抗体共同孵育,可使成纤维细胞的[³H]胸腺嘧啶核苷掺入量从7800±1000 cpm增加至18900±800 cpm(P<0.01),平滑肌细胞的掺入量从9800±1200 cpm增加至17200±1100 cpm(P<0.01)。胰岛素可作为有丝分裂原替代Sm-C且不与该抗体结合,与抗体共同孵育时可刺激DNA合成,从而排除了非特异性细胞毒性的可能性。这些结果强化了这样一种假说,即这两种细胞类型在体外DNA合成的速率与其产生类生长调节素肽的能力直接相关。它们进一步支持了类生长调节素肽的细胞产生是生长调节自分泌模型的实例这一观点。