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1,25-(OH)₂-维生素D₃对未转化和转化的C3H/10T1/2细胞克隆中同源受体和c-myc诱导作用的时间进程研究

Time-course study of 1,25-(OH)2-vitamin D3 induction of homologous receptor and c-myc in nontransformed and transformed C3H/10T1/2 cell clones.

作者信息

Paatero G I, Trydal T, Karlstedt K A, Aarskog D, Lillehaug J R

机构信息

Department of Biology, Abo Akademi University, Finland.

出版信息

Int J Biochem. 1994 Mar;26(3):367-74. doi: 10.1016/0020-711x(94)90056-6.

DOI:10.1016/0020-711x(94)90056-6
PMID:8187933
Abstract
  1. We have used 1,25-dihydroxyvitamin D3 (1,25-(OH)2D3) to investigate autoregulation of homologous receptor and the control of c-myc mRNA and protein expression in C3H/10T1/2 cells. 2. 10 nM 1,25-(OH)2D3 stimulated 1,25-(OH)2D3 receptor (VDR) synthesis in both non-transformed C3H/10T1/2 Cl 8 and in chemically transformed C3H/10T1/2 Cl 16 cells within 4 hr of treatment. Maximal induction was observed between 8 and 24 hr. 3. Two VDR mRNA transcripts, 2.7 and 4.8 kb, were present in both cell types. There were parallel changes in VDR specific mRNA levels and cellular VDR concentration in the C3H/10T1/2 Cl 8 cells indicating that the increase in receptor concentrations was dependent on de novo mRNA synthesis. 4. The increase in VDR mRNA concentration in the chemically transformed C3H/10T1/2 Cl 16 cells was maximal already at 4 hr, preceding the maximal increase in receptor concentration by 4-6 hr. 5. Analysis of c-myc mRNA levels also showed cell line specificity. 6. The c-myc mRNA level increased 2.1-fold with 10 nM 1,25-(OH)2D3 treatment in C3H/10T1/2 Cl 8 cells after 12 hr while the C3H/10T1/2 Cl 16 cells had maximal c-myc mRNA level after 1 hr. 7. The relative amount of c-myc mRNA remained higher than that of unstimulated controls the next 10-12 hr in C3H/10T1/2 Cl 16 cells. 8. The c-myc protein levels were not affected by 1,25-(OH)2D3 treatment in either cell line as detected by Western blot analysis. 9. Our data suggest that 1,25-(OH)2D3 mediated induction of VDR does not require prior c-myc protein synthesis in the C3H/10T1/2 cells.(ABSTRACT TRUNCATED AT 250 WORDS)
摘要
  1. 我们使用1,25 - 二羟基维生素D3(1,25-(OH)2D3)来研究同源受体的自身调节以及C3H/10T1/2细胞中c - myc mRNA和蛋白质表达的调控。2. 10 nM的1,25-(OH)2D3在处理4小时内刺激了未转化的C3H/10T1/2 Cl 8细胞和化学转化的C3H/10T1/2 Cl 16细胞中1,25-(OH)2D3受体(VDR)的合成。在8至24小时之间观察到最大诱导。3. 两种VDR mRNA转录本,2.7 kb和4.8 kb,存在于两种细胞类型中。C3H/10T1/2 Cl 8细胞中VDR特异性mRNA水平和细胞VDR浓度存在平行变化,表明受体浓度的增加依赖于从头合成mRNA。4. 化学转化的C3H/10T1/2 Cl 16细胞中VDR mRNA浓度在4小时时已达到最大值,比受体浓度的最大增加提前4至6小时。5. 对c - myc mRNA水平的分析也显示出细胞系特异性。6. 在C3H/10T1/2 Cl 8细胞中,10 nM的1,25-(OH)2D3处理12小时后,c - myc mRNA水平增加了2.1倍,而C3H/10T1/2 Cl 16细胞在1小时后达到最大c - myc mRNA水平。7. 在接下来的10 - 12小时内,C3H/10T1/2 Cl 16细胞中c - myc mRNA的相对量仍高于未刺激的对照。8. 通过蛋白质印迹分析检测,两种细胞系中c - myc蛋白质水平均不受1,25-(OH)2D3处理的影响。9. 我们的数据表明,在C3H/10T1/2细胞中,1,25-(OH)2D3介导的VDR诱导不需要先前的c - myc蛋白质合成。(摘要截短至250字)

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