Wax S D, Tsao L, Lieb M E, Fallon J T, Taubman M B
Department of Medicine, Brookdale Center for Molecular Biology, Mount Sinai School of Medicine, New York, New York, USA.
Lab Invest. 1996 Apr;74(4):797-808.
We have previously reported the identification of a novel cDNA, SM-20, whose corresponding mRNA levels are regulated by growth factors in rat aortic smooth muscle cell (SMC) culture. Affinity-purified polyclonal and monoclonal antibodies were made against a 230 amino acid region of the SM-20 putative peptide expressed in the bacterial vector, pET-3b. Western blot analyses of rat and human SMC lysates detected a single protein species of approximately 40 kd. SM-20 was not detected in concentrates of the culture medium. By immunohistochemistry, SM-20 was localized to filaments in the cytoplasm of cultured SMC. Like SM-20 mRNA, levels of SM-20 protein were increased 1-3 hours after serum stimulation of rat aortic SMC. In rat tissues, SM-20 antigen was detected in abundance in smooth, cardiac, and skeletal muscle. SM-20 was also detected in some epithelial cells of the kidney, pancreas, gastrointestinal tract, and skin, but was not found in the parenchyma of the liver, spleen, or testis. In the rat arterial wall, SM-20 antigen was restricted to the SMC of the media and, after balloon arterial injury, was found most abundantly in the neointima. In human atherosclerotic coronary arteries, SM-20 antigen predominated in the SMC of the intimal plaques and was not detected in macrophages, endothelium, or adventitial cells. Thus, SM-20 encodes a protein which serves as a novel SMC-specific marker within the vessel wall.
我们先前报道了一种新型cDNA即SM - 20的鉴定,其相应的mRNA水平在大鼠主动脉平滑肌细胞(SMC)培养中受生长因子调控。针对在细菌载体pET - 3b中表达的SM - 20推定肽的230个氨基酸区域制备了亲和纯化的多克隆和单克隆抗体。对大鼠和人SMC裂解物的蛋白质印迹分析检测到一种约40 kd的单一蛋白质。在培养基浓缩物中未检测到SM - 20。通过免疫组织化学,SM - 20定位于培养的SMC细胞质中的细丝。与SM - 20 mRNA一样,大鼠主动脉SMC血清刺激后1 - 3小时,SM - 20蛋白水平升高。在大鼠组织中,在平滑肌、心肌和骨骼肌中大量检测到SM - 20抗原。在肾脏、胰腺、胃肠道和皮肤的一些上皮细胞中也检测到SM - 20,但在肝实质、脾或睾丸中未发现。在大鼠动脉壁中,SM - 20抗原局限于中膜的SMC,在球囊动脉损伤后,在内膜中含量最丰富。在人动脉粥样硬化冠状动脉中,SM - 20抗原在内膜斑块的SMC中占主导地位,在巨噬细胞、内皮细胞或外膜细胞中未检测到。因此,SM - 20编码一种蛋白质,该蛋白质可作为血管壁内一种新型的SMC特异性标志物。