QUANTITATIVE POLYMERASE CHAIN-REACTION OF LYSYL OXIDASE MESSENGER-RNA IN MALIGNANTLY TRANSFORMED HUMAN CELL-LINES DEMONSTRATES THAT THEIR LOW LYSYL OXIDASE ACTIVITY IS DUE TO LOW QUANTITIES OF ITS MESSENGER-RNA AND LOW-LEVELS OF TRANSCRIPTION OF THE RESPECTIVE GENE
QUANTITATIVE POLYMERASE CHAIN-REACTION OF LYSYL OXIDASE MESSENGER-RNA IN MALIGNANTLY TRANSFORMED HUMAN CELL-LINES DEMONSTRATES THAT THEIR LOW LYSYL OXIDASE ACTIVITY IS DUE TO LOW QUANTITIES OF ITS MESSENGER-RNA AND LOW-LEVELS OF TRANSCRIPTION OF THE RESPECTIVE GENE
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DOI：
10.1074/jbc.270.37.21590
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发表时间：
1995-09-15
影响因子：
4.8
通讯作者：
KIVIRIKKO, KI
中科院分区：
文献类型：
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作者：
HAMALAINEN, ER;KEMPPAINEN, R;KIVIRIKKO, KI
赖氨酰氧化酶（EC 1.4.3.13）是一种细胞外铜氨基氧化酶，通过催化某些赖氨酸和羟赖氨酸残基中ε-氨基的氧化脱氨基作用，引发胶原蛋白和弹性蛋白的交联。我们在这里开发了一种定量赖氨酰氧化酶mRNA的聚合酶链反应（PCR）方法，其中合成的RNA用作与靶mRNA共扩增的内标。当通过北方印迹分析研究赖氨酰氧化酶mRNA的量时，和当通过定量PCR方法测定赖氨酰氧化酶mRNA分子的数目时，发现在各种恶性转化的细胞系中相对于对照细胞细颗粒明显较低，定量PCR指示值为对照中的值的约2-10%。在转化细胞和对照之间的β-肌动蛋白mRNA分子的数量没有发现差异。核径流实验表明，赖氨酰氧化酶mRNA分子数量的减少，如果不是全部的话，大部分可以解释为各自基因的转录减少。
Lysyl oxidase (EC 1.4.3.13), an extracellular copper amino oxidase, initiates the cross-linking of collagens and elastin by catalyzing oxidative deamination of the epsilon-amino group in certain lysine and hydroxylysine residues. We developed here a polymerase chain reaction (PCR) method for the quantification of lysyl oxidase mRNA in which a synthetic RNA is used as an internal standard for coamplification with the targeted mRNA. The amount of lysyl oxidase mRNA when studied by Northern blot analysis and the number of lysyl oxidase mRNA molecules when determined by the quantitative PCR method were found to be markedly low in various malignantly transformed cell lines relative to control cell fines, quantitative PCR indicating values of about 2-10% of those in the controls. No difference was found in the number of beta-actin mRNA molecules between the transformed cells and the controls. Nuclear runoff experiments indicated that most if not all of the decrease in the number of lysyl oxidase mRNA molecules can be explained by diminished transcription of the respective gene.