Functional Analysis of Zebrafish (Danio rerio) IGFBP-1 and IGFBP-2 and Biological Activities of Fish Recombinant Protein in Inhibition of Tumor Cell Differentiation

斑馬魚類胰島素生長因子第一型與第二型之功能與重組蛋白抑制癌細胞分化分析

陳志毅(Jyh-Yih Chen) ； 陳健祺(Jian-Chyi Chen) ； 吳金洌(Jen-Leih Wu)

《臺灣水產學會刊》 31卷4期 (2004/12) Pp. 295-310

https://doi.org/10.29822/JFST.200412.0006

摘要

這個實驗的目標是對類胰島素生長因子結合蛋白第一型與第二型在飢餓，胰島素生長因子，胰島素，和生長激素在內分泌學上的調控影響研究。類胰島素生長因子結合蛋白第一型與第二型利用定量PCR來偵測其上的表現量之差異。類胰島素生長因子結合蛋白第一型可以在肝臟、鰭，卵巢、心臟上表現高濃度，類胰島素生長因子結合蛋白第二型可以在卵巢與肌肉上表現高濃度。饑餓三週後，類胰島素生長因子結合蛋白第一型與第二型的基因表現量會昇高。注射胰島素48小時後可以發現類胰島素生長因子結合蛋白第二型基因表現具有差異性。利用MTT的分析類胰島素生長因子結合蛋白第一型與第二型的重組蛋白作用在NIH3T3，HT1080，B16F10，HeLa，MCF-7的細胞上，可以發現具有抑制這些所被測試的細胞株。這些研究成果暗示類胰島素生長因子結合蛋白扮演很重要的角色在不同的細胞型態與器官中。

關鍵字

類胰島素生長因子結合蛋白 ； 饑餓 ； 荷爾蒙調控

並列摘要

The aim of this work was to study the influences of the endocrine balance during starvation, insulin-like growth factors, insulin, and growth hormone (GH) on the regulation of insulin-like growth factor binding protein-1 and -2. mRNA concentrations of IGFBP-1 and -2 were assayed by comparative reverse-transcription polymerase chain reaction (PCR). RT-PCR detected high expressions of IGFBP-1 in The liver, fin, ovary, and heart. IGFBP-2 was highly expressed in the ovary and muscle. Starvation results show that IGFBP-1 and -2 mRNAs had elevated expression levels after 3 weeks of fasting. The injected hormone experiment showed that there were significant differences in the expressions of IGFBP-2 mRNA 48h after injection of insulin. Recombinant zebrafish IGFBP-1 and -2 polypeptide bioactivities on the growth-promoting activity of the cell lines NIH3T3, HT1080, B16F10, HeLa, and MCF-7 were determined by an MTT assay. These changes were measured using MTT to assess mitochondrial reductase activity. Recombinant IGFBP-1 and -2 protein bioactivities showed inhibition in all cell lines tested. Despite such small differences, these results indicate that both their structure and involvement in regulation by growth factors of IGFBP-1 and -2 proteins appear to have been remarkably conserved. These results imply that the biological actions of IGFBPs in zebrafish may play an important role in different cell types and organ systems.

並列關鍵字

insulin-like growth factor binding protein ； starvation ； hormonal regulation ； recombinant protein ； tumor suppression