RETINOIC ACID STIMULATES INTERSTITIAL COLLAGENASE MESSENGER-RIBONUCLEIC-ACID IN OSTEOSARCOMA CELLS
RETINOIC ACID STIMULATES INTERSTITIAL COLLAGENASE MESSENGER-RIBONUCLEIC-ACID IN OSTEOSARCOMA CELLS
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DOI：
10.1210/en.135.6.2542
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发表时间：
1994-12-01
期刊：
影响因子：
4.8
通讯作者：
QUINN, CO
中科院分区：
文献类型：
--
作者：
CONNOLLY, TJ;CLOHISY, JC;QUINN, CO
大鼠成骨性骨肉瘤细胞系UMR 106-01在维甲酸(RA)作用下可分泌间质胶原酶。Northern印迹分析表明，RA在6h可引起胶原酶信使RNA(MR NA)的表达增加，2 4h达最大值(为基础水平的2 0.5倍)，72h下降至基础水平，这种刺激作用呈剂量依赖关系，在5×10(-7)M RA时反应最大。核连续分析显示，RA治疗后12-24小时，胶原酶mRNA转录速率增加了20倍以上。放线菌酮阻断RA对胶原酶mRNA的刺激，证明了从头合成蛋白质的必要性。RA不仅导致胶原酶分泌增加，而且已知还会减少UMR 106-01细胞的胶原合成。在这项研究中，胶原酶mRNA的增加伴随着α(1)(I)前胶原mRNA水平的下降，在24小时达到最大(下降70%)，并在72小时恢复到接近对照的水平。核连续分析表明，α(1)(I)前胶原表达的下降没有统计学意义。RA不降低α(1)(I)前胶原mRNA的稳定性(计算t(1/2)=8.06+/-0.30和9.01+/-0.62h)。然而，转录和稳定性可能共同导致了稳定的α(1)(I)前胶原mRNA的主要下降。放线菌酮治疗抑制了基础水平的α(1)(I)前胶原mRNA的积累，表明需要持续的蛋白质合成来维持该基因的基础表达。
The rat osteoblastic osteosarcoma cell line UMR 106-01 secretes interstitial collagenase in response to retinoic acid (RA). The present study demonstrates by Northern blot analysis that RA causes an increase in collagenase messenger RNA (mRNA) at 6 h, which is maximal at 24 h (20.5 times basal) and declines toward basal level by 72 h. This stimulation is dose dependent, with a maximal response at 5 x 10(-7) M RA. Nuclear run-on assays show a greater than 20-fold increase in the rate of collagenase mRNA transcription between 12-24 h after RA treatment. Cycloheximide blocks RA stimulation of collagenase mRNA, demonstrating the need for de novo protein synthesis. RA not only causes an increase in collagenase secretion, but is known to decrease collagen synthesis in UMR 106-01 cells. In this study, the increase in collagenase mRNA is accompanied by a concomitant decrease in the level of alpha(1)(I) procollagen mRNA, which is maximal at 24 h (70% decrease), with a return to near-control levels by 72 h. Nuclear run-on assays demonstrated that the decrease in alpha(1)(I) procollagen expression does not have a statistically significant transcriptional component. RA did not statistically decrease the stability of alpha(1)(I) procollagen mRNA (calculated t(1/2) = 8.06 +/- 0.30 and 9.01 +/- 0.62 h in the presence and absence of RA, respectively). However, transcription and stability together probably contribute to the major decrease in stable alpha(1)(I) procollagen mRNA observed. Cycloheximide treatment inhibits basal level alpha(1)(I) procollagen mRNA accumulation, demonstrating the need for on-going protein synthesis to maintain basal expression of this gene.