Relative quantification of angiotensin-converting enzyme mRNA in human smooth muscle cells, monocytes, and lymphocytes by the polymerase chain reaction.
Relative quantification of angiotensin-converting enzyme mRNA in human smooth muscle cells, monocytes, and lymphocytes by the polymerase chain reaction.
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通过聚合酶链式反应对人平滑肌细胞、单核细胞和淋巴细胞中血管紧张素转换酶 mRNA 进行相对定量。
DOI：
10.1006/abio.1994.1260
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发表时间：
1994
影响因子：
2.9
通讯作者：
Lanzillo,JJ
中科院分区：
文献类型：
--
作者：
Aschoff,JM;Lazarus,D;Fanburg,BL;Lanzillo,JJ
建立了一种在100个细胞中相对定量血管紧张素转换酶（ACE）mRNA的方法。将总RNA逆转录为cDNA后，使用两组引物进行多重聚合酶链反应，同时扩增ACE cDNA和内标磷酸甘油醛脱氢酶（GAPDH）的cDNA。通过调节引物对浓度，ACE和GAPDH都以恒定的效率扩增。巨噬细胞样U937组织细胞性淋巴瘤细胞组成型表达ACE mRNA。新鲜分离的人单核细胞最初不表达ACE mRNA，但培养4天后，其表达量为U937细胞的8%。佛波酯刺激后，单核细胞转录ACE的水平与U937细胞相当。人平滑肌细胞的ACE mRNA比4天单核细胞高9倍，但比U937细胞低30%。相反，混合的淋巴细胞群体没有ACE mRNA。
A method was developed for relative quantification of angiotensin-converting enzyme (ACE) mRNA in as few as 100 cells. After reverse transcription of total RNA to cDNA, multiplexed polymerase chain reaction with two sets of primers amplified ACE cDNA and that of an internal standard glyceraldehyde phosphate dehydrogenase (GAPDH) simultaneously. By adjusting primer pair concentrations, both ACE and GAPDH were amplified with constant efficiencies. Macrophage-like U937 histiocytic lymphoma cells expressed ACE mRNA constitutively. Freshly isolated human monocytes did not express ACE mRNA initially, but after 4 days in culture had 8% of the amount found in U937 cells. After phorbol ester stimulation, monocytes transcribed ACE at levels comparable to U937 cells. Human smooth muscle cells had ninefold more ACE mRNA than 4-day monocytes, but 30% less than U937 cells. In contrast, a mixed population of lymphocytes was devoid of ACE mRNA.