Stockert R J
Department of Medicine, Marion Bessin Liver Research Center, Albert Einstein College of Medicine, Bronx, New York 10461.
J Biol Chem. 1993 Sep 15;268(26):19540-4.
The mechanism of regulated expression of the asialoglycoprotein receptor (ASGR) by cAMP was investigated in the human hepatoblastoma cell line, HepG2. Incubation of HepG2 cells with the cell-permeant 8-bromo-cAMP or induction of intracellular cAMP with forskolin reduced receptor expression in confluent HepG2 cultures. Immunoblot analysis established that this reduction of receptor activity was due to a reduction of expression of both ASGR subunit polypeptides H1 and H2. Estimates of the steady-state levels of H1- and H2-related mRNA by Northern blot analysis indicated that reduced ASGR expression was a result of a decrease in gene transcript number. By a combination of run-on and mRNA turnover studies, it was suggested that this reduction of ASGR-related mRNA resulted from its destabilization induced by 8-bromo-cAMP. The effect of 8-bromo-cAMP appears not to be limited to ASGR expression as a rapid reduction in the albumin mRNA was also observed. In contrast, both the beta-actin and glyceraldehyde-3-phosphate dehydrogenase mRNA levels were elevated by exposure to 8-bromo-cAMP.
在人肝癌细胞系HepG2中研究了环磷酸腺苷(cAMP)对去唾液酸糖蛋白受体(ASGR)表达调控的机制。用可透过细胞的8-溴环磷酸腺苷孵育HepG2细胞,或用福司可林诱导细胞内cAMP,均可降低汇合的HepG2培养物中的受体表达。免疫印迹分析表明,受体活性的这种降低是由于ASGR亚基多肽H1和H2的表达均减少所致。通过Northern印迹分析对H1和H2相关mRNA的稳态水平进行估计,结果表明ASGR表达降低是基因转录本数量减少的结果。通过连续转录和mRNA周转研究相结合的方法,提示ASGR相关mRNA的这种减少是由8-溴环磷酸腺苷诱导其不稳定所致。8-溴环磷酸腺苷的作用似乎并不局限于ASGR的表达,因为还观察到白蛋白mRNA迅速减少。相反,β-肌动蛋白和甘油醛-3-磷酸脱氢酶mRNA水平在暴露于8-溴环磷酸腺苷后升高。