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Differential expression of cGMP‐inhibited cyclic nucleotide phosphodiesterases in human hepatoma cell lines

机译:cGMP抑制的环核苷酸磷酸二酯酶在人肝癌细胞系中的差异表达

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>PDE3 or cGMP-inhibited cyclic nucleotide phosphodiesterase (cGI PDE) activity was detected in homogenates of HepG2, Hep3B and HuH7, but not SK-Hep-1, human hepatoma cells. In HepG2 and Hep3B cells PDE3 activity was found predominantly in particulate fractions; in HuH7, in both particulate and supernatant fractions. cDNAs encoding two human PDE3s (an ‘adipocyte’ type, HcGIP1),and a ‘cardiovascular’ type, HcGIP2) have been cloned. HcGIP1 cDNA hybridized strongly with poly(A)+ RNA species from HepG2 and Hep3B. Both HcGIPI and HcGIP2 mRNAs were expressed in Hep3B and HuH7 cells. The nucleotide sequence of an ∼300-bp cDNA fragment, isolated after RT-PCR cloning from HepG2 RNA, was identical to a sequence within the conserved domain of HcGIP1 cDNA, consistent with the presence of HcGIPI mRNA in HepG2 cells.
机译:在人肝癌HepG2,Hep3B和HuH7的匀浆物中检测到pDE3或cGMP抑制的环核苷酸磷酸二酯酶(cGI PDE)活性。在HepG2和Hep3B细胞中,PDE3活性主要存在于颗粒级分中。在HuH7中,无论是颗粒级分还是上清级分。已克隆了编码两种人类PDE3(一种“脂肪细胞”型,HcGIP1)和一种“心血管”型,HcGIP2的cDNA。 HcGIP1 cDNA与HepG2和Hep3B的poly(A) + RNA种类强烈杂交。 HcGIPI和HcGIP2 mRNA均在Hep3B和HuH7细胞中表达。从HepG2 RNA进行RT-PCR克隆后分离出的约300 bp cDNA片段的核苷酸序列与HcGIP1 cDNA保守域内的序列相同,这与HepG2细胞中HcGIPI mRNA的存在一致。

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