Expression and promoter-alternation of osteoblastic insulin-like growth factors

成骨细胞胰岛素样生长因子的表达和启动子改变

基本信息

批准号：
06671854
负责人：
TAKAHASHI Kojiro
金额：
$ 1.41万
依托单位：
Okayama University
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (C)
财政年份：
1994
资助国家：
日本
起止时间：
1994 至 1995
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-06671854/
关键词：
Insulin-like growth factor II Insulin-like growth factor I Osteoblastic cell Chondrosarcoma Chondrocytes Transcriptional promoter RT-PCR RNase Protection Assay Insulin-like growth factor-II Insulim-like growth factor-I 軟骨様細胞転写調節 I

项目摘要

Using a chondrocytic cell line HCS-2/8 derived from human chondrosarcoma, the quantitative analytical method adapting the RNase protection Assay (RPA) was developped in order to examine the expression and promoter-alternation of insulin-like growth factor genes. The probe of the RPA experiments was constructed for the promoters 2P-1,2P-3 and 2P-4 of IGF-2 and the promoters 1P-1 and 1P-2 of IGF-1, but the promoter 2P-2 of IGF-2 failed to be constructed because the transcriptional product was very little in HCS-2/8. Now, we continue to investigate the determination of detectable limitation of the RPA method using their probes, and we will retry to construct the probe asfter the accumulation of RT-PCR product for 2P-2 transcript.During the present developmental investigations, we found some new facts as following :1.The simultaneous expression of four promoters of IGF-2 were induced in HCS-2/8 and human normal chondrocytes.2.A defect of four base pair in the promoter 2P-4 transcript of IGF-2 in HCS-2/8 was observed at the 3'-end of exon 6, at which the position is upstream of 9 to 6 base from the translation-starting point. This defect may be due to the alternative splicing specific to the chondrosarcoma.3.Comparing with HCS-2/8 and human normal chondrocytes, we found a parallel correlation between IGF-2 and H19 (tumor reppressor gene) expressions.4.The restriction fragment length polymorphism of IGF-2 gene in HCS-2/8 suggested that only the paternal allele (s) existed at the genomic level, but the maternal imprinting allele was absent in the chondrosarcoma. This perhaps correlate with the tumor formation.

使用源自人软骨肉瘤的软骨细胞系HCS-2/8，开发了适应RNase保护测定法（RPA）的定量分析方法，以检查胰岛素样生长因子基因的表达和启动子抗性。为IGF-2的启动子2p-1,2p-3和2p-4构建了RPA实验的探针，IGF-1的启动子1P-1和1P-2构建了IGF-1的启动子1P-1和1P-2，但是IGF-2的启动子2P-2无法构造，因为HCS-2/8中的转录产物很少。现在，我们继续使用其探针调查RPA方法可检测到的限制，我们将重试构建对2P-2转录的RT-PCR产物的积累的探测。在当前的发展研究中，我们发现了一些新事实，如以下一些新事实：1。IGF-2的四四个基本范围和人类的四个基本范围。在HCS-2/8中IGF-2的启动子2p-4转录本中观察到外显子6的3末端，该位置从转换启动点开始在9至6个基础上。该缺陷可能是由于软骨肉瘤的替代剪接引起的。3.com与HCS-2/8和人类正常软骨细胞相比，我们发现IGF-2和H19和H19和H19（肿瘤reppressor reppressor基因）的表达之间存在平行相关。4。限制性的限制长度碎片的限制性碎片eigf-2基因igf-2基因在hcf-2 in h hc in h hcc in h hccy in hc in h hcc in h h h h h h h h h h h h h h h h h h h h h h h h h h h h h h h h h h h h h h h h h h h h i Nep的。。。。。。基因组水平，但是软骨肉瘤中不存在母体烙印等位基因。这可能与肿瘤形成有关。