ISOLATION OF GENE MARKERS OF DIFFERENTIATED AND PROLIFERATING VASCULAR SMOOTH-MUSCLE CELLS
ISOLATION OF GENE MARKERS OF DIFFERENTIATED AND PROLIFERATING VASCULAR SMOOTH-MUSCLE CELLS
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DOI：
10.1161/01.res.73.1.193
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发表时间：
1993-07-01
影响因子：
20.1
通讯作者：
METCALFE, JC
中科院分区：
文献类型：
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作者：
SHANAHAN, CM;WEISSBERG, PL;METCALFE, JC
为了分离分化和增殖的血管平滑肌细胞（VSMCs）的特异性标志物，我们使用了差异cDNA筛选技术，从培养的大鼠主动脉VSMCs的RNA。通过北方分析确定所有分离的cDNA表达的组织特异性。我们分离出7种不同的cDNA，它们在新鲜分散的、分化的主动脉VSMCs中的表达比去分化的晚期传代细胞更强。这些是原弹性蛋白的cDNA，基质蛋白; α-平滑肌（SM）肌动蛋白，γ-SM肌动蛋白，钙调蛋白和受磷蛋白，这些都是与分化的VSMC的收缩功能相关的蛋白质; SM 22 α，一种功能未知的平滑肌特异性蛋白质，和CHIP 28，一种推定的膜通道蛋白，在其他SM组织中不高表达，因此可能是一种新的VSMC标记物。还分离出两个在晚期传代去分化的VSMCs中优先表达的cDNA。这些是骨桥蛋白和基质Gla蛋白（MGP）的cDNA。与CHIP 28一样，MGP在主动脉VSMCs中强烈表达，但在含有SM细胞的其他类型的组织中不表达，这表明两者在血管组织中具有特异性功能。骨桥蛋白和MGP以前都从发育中的骨中分离出来。它们在增殖的VSMCs中的表达表明它们可能参与调节血管病变中常见的钙化。所获得的cDNA组扩展了可用于通过原位杂交在体内鉴定VSMC和表征其表型的DNA探针的范围。因此，它们应该有助于分析血管病变发展过程中的基因表达。
To isolate specific markers of both differentiated and proliferating vascular smooth muscle cells (VSMCs), we used the technique of differential cDNA screening using RNA from cultured rat aortic VSMCs. The tissue specificity of expression of all of the cDNAs isolated was determined by Northern analysis. We isolated seven distinct cDNAs that were more strongly expressed in freshly dispersed, differentiated, aortic VSMCs compared with dedifferentiated late-passage cells. These were the cDNAs for tropoelastin, a matrix protein; alpha-smooth muscle (SM) actin, gamma-SM actin, calponin, and phospholamban, which are all proteins associated with the contractile function of differentiated VSMCs; SM22alpha, a smooth muscle-specific protein of unknown function, and CHIP28, a putative membrane channel protein that is not highly expressed in other SM tissues and may therefore be a new VSMC marker. Two cDNAs that were expressed preferentially in late-passage dedifferentiated VSMCs were also isolated. These were the cDNAs for osteopontin and matrix Gla protein (MGP). Like CHIP28, MGP was strongly expressed in aortic VSMCs but not in other types of tissues containing SM cells, suggesting that both have specific functions in vascular tissue. Osteopontin and MGP have both previously been isolated from developing bone. Their expression in proliferating VSMCs suggests that they may be involved in regulating the calcification that commonly occurs in vascular lesions. The set of cDNAs obtained extends the range of DNA probes that are available for identifying VSMCs and characterizing their phenotype in vivo by in situ hybridization. Therefore, they should aid in the analysis of gene expression during the development of vessel lesions.