Differential effects of extracellular matrix proteins on human airway smooth muscle cell proliferation and phenotype
Differential effects of extracellular matrix proteins on human airway smooth muscle cell proliferation and phenotype
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DOI：
10.1165/ajrcmb.23.3.3990
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发表时间：
2000-09-01
影响因子：
6.4
通讯作者：
Lee, TH
中科院分区：
文献类型：
--
作者：
Hirst, SJ;Twort, CHC;Lee, TH
成熟的气道平滑肌细胞的特征在于低增殖指数和表达收缩标记蛋白，如平滑肌α-肌动蛋白（sm-alpha-actin）、钙调蛋白和平滑肌肌球蛋白重链（sm-MHC）。在本研究中，定义的细胞外基质（ECM）成分进行了检查，对有丝分裂原刺激的，培养的人气道平滑肌细胞的增殖和表型状态。结果表明，尽管细胞在用1 - 100 μ g/ml纤连蛋白（FN）、胶原I（Col I）、层粘连蛋白（LN）或基质胶预包被的平板上粘附和铺展，但它们随后的增殖反应在性质上不同。与塑料上的细胞相比，FN和Col I增强了对血小板衍生生长因子（PDGF）-BB或α-凝血酶的增殖反应。LN，但是，减少有丝分裂原刺激的增殖。在基质胶上培养的细胞中发现了类似的减少。ECM底物对收缩表型的影响通过使用免疫细胞化学和流式细胞术方法检查sm-alpha-肌动蛋白、sm-MHC和钙调蛋白的细胞表达来确定。大约75%的PDGF-BB刺激的细胞，培养在LN或Matrigel，表达sm-or-肌动蛋白，钙调蛋白，和sm-MHC，但只有8至10%的Ki 67核抗原增殖标记染色。相比之下，超过75%的FN或Col I培养的细胞Ki 67抗原阳性，但只有20%的收缩蛋白阳性。sm-alpha-肌动蛋白和DNA含量的流式细胞术分析证实了免疫细胞化学的结果，并表明，观察到的减少sm-alpha-肌动蛋白含量培养后FN或Col I，LN和基质胶相比，发生在大多数的细胞群体，支持双向表型调制。总之，数据表明ECM底物调节培养的人气道平滑肌细胞的增殖和表型。
Mature airway smooth muscle cells are characterized by a low proliferative index and expression of contractile marker proteins such as smooth muscle alpha-actin (sm-alpha-actin), calponin, and smooth muscle myosin heavy chain (sm-MHC). In the present study, defined extracellular matrix (ECM) components were examined on the proliferative and phenotypic status of mitogen-stimulated, cultured human airway smooth muscle cells. The results demonstrate that although cells adhered and spread on plates precoated with (1 to 100 (mu g/ml) of fibronectin (FN), collagen I (Col I), laminin (LN), or Matrigel, their subsequent proliferative response varied qualitatively. FN and Col I enhanced proliferation in response to either platelet-derived growth factor (PDGF)-BB or alpha-thrombin, compared with cells on plastic. LN, however, reduced mitogen-stimulated proliferation. A similar reduction was found in cells cultured on Matrigel. The effect of ECM substrates on contractile phenotype was determined by examining cellular expression of sm-alpha-actin, sm-MHC, and calponin using immunocytochemical and flow cytometric methods. Approximately 75% of PDGF-BB-stimulated cells, cultured on LN or Matrigel, expressed sm-or-actin, calponin, and sm-MHC, but only 8 to 10% stained for the Ki67 nuclear antigen proliferation marker. In contrast, more than 75% of cells cultured on FN or Col I were positive for Ki67 antigen, but only 20% were positive for contractile proteins. Flow cytometric analysis of sm-alpha-actin and DNA content confirmed the immunocytochemical findings and showed that the observed reduction in sm-alpha-actin content after culture on FN or Col I, compared with LN and Matrigel, occurred in the majority of the cell population, supporting bidirectional phenotype modulation. Overall, the data suggest that ECM substrates modulate both proliferation and phenotype of human airway smooth muscle cells in culture.