Brinckerhoff C E, Plucinska I M, Sheldon L A, O'Connor G T
Biochemistry. 1986 Oct 21;25(21):6378-84. doi: 10.1021/bi00369a006.
As part of our studies on the mechanisms controlling the synthesis of the neutral proteinase collagenase by rabbit synovial cells, we used a cDNA clone to measure total collagenase mRNA levels and to determine mRNA half-life. Phorbol myristate acetate was used to induce collagenase synthesis while all-trans-retinoic acid and dexamethasone were used to inhibit it. Cells stimulated with phorbol myristate acetate contained substantial amounts of collagenase mRNA, but cells treated with all-trans-retinoic acid or dexamethasone contained decreased amounts of collagenase mRNA which correlated well with levels of collagenase protein. Studies on mRNA half-life showed that the t1/2 for total poly(A+) RNA was about 25 h, while that of collagenase varied from as short as 12 h to as long as 36 h. The half-life was not affected by treatment with all-trans-retinoic acid or dexamethasone but was affected by the level of induction of collagenase mRNA: the greater the amount of collagenase mRNA induced, the longer the t1/2. We conclude that our data are consistent with the hypothesis that retinoic acid and dexamethasone act at the level of transcription to decrease collagenase production and the increased level of collagenase mRNA resulting from stimulation with phorbol esters is, in part, due to increased stability of the induced collagenase mRNA.
作为我们对兔滑膜细胞中性蛋白酶胶原酶合成调控机制研究的一部分，我们使用一个cDNA克隆来测量总胶原酶mRNA水平并确定mRNA半衰期。佛波酯用于诱导胶原酶合成，而全反式维甲酸和地塞米松则用于抑制其合成。用佛波酯刺激的细胞含有大量的胶原酶mRNA，但用全反式维甲酸或地塞米松处理的细胞中胶原酶mRNA含量降低，这与胶原酶蛋白水平密切相关。对mRNA半衰期的研究表明，总聚腺苷酸（poly(A+)）RNA的半衰期约为25小时，而胶原酶的半衰期则从短至12小时到长达36小时不等。半衰期不受全反式维甲酸或地塞米松处理的影响，但受胶原酶mRNA诱导水平的影响：诱导的胶原酶mRNA量越大，半衰期越长。我们得出结论，我们的数据与以下假设一致，即维甲酸和地塞米松在转录水平起作用以减少胶原酶的产生，并且佛波酯刺激导致的胶原酶mRNA水平升高部分归因于诱导的胶原酶mRNA稳定性增加。
