Isolation of marmoset P450c17 cDNA and gene regulation in vitro.
Isolation of marmoset P450c17 cDNA and gene regulation in vitro.
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狨猴 P450c17 cDNA 的分离和体外基因调控。
DOI：
10.1081/erc-200044018
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发表时间：
2004
影响因子：
2.1
通讯作者：
Bird,IanM
中科院分区：
文献类型：
--
作者：
Pattison,JChristina;Conley,AlanJ;Bird,IanM
绒猴在肾上腺发育期间表达胎儿区，但在成年期无法产生大量的C19类固醇。然而，尚不清楚P450 c17调控是否与人类/恒河猴不同或一级序列是否发生改变。为此，我们分离了绒猴和恒河猴的肾上腺皮质细胞，并用已知的P450 c17表达调节剂处理细胞48小时。P450 c17蛋白随Forskolin（F）处理而增加，但仅被AII（A）和TPA（T）略微抑制。联合A + F和T + F显著消融F反应。F处理后，皮质醇水平（EIA）升高，A和T抑制。联合给药可部分抑制F诱导的反应。然后使用RT-PCR/TA克隆从肾上腺中分离绒猴和恒河猴P450 c17 cDNA的蛋白编码区。绒猴P450 c17有一个氨基酸缺失，但与人和恒河猴的cDNA序列同源性分别为90.6%和91.4%，与人和恒河猴的预测AA序列同源性分别为82.4%和85%。由于绒猴肾上腺皮质细胞表现出与恒河猴相似的内分泌功能，成年绒猴肾上腺中17，20-裂解酶活性受损可能部分是由于初级序列的差异。
Marmosets express a fetal zone during adrenal development but fail to produce significant amounts of C19 steroids in adulthood. It is not clear however if P450c17 regulation is different from that in humans/rhesus or the primary sequence is altered. To this end we isolated marmoset and rhesus adrenocortical cells, and treated the cells with known regulators of P450c17 expression for 48 h. P450c17 protein increased with Forskolin (F) treatment, but was marginally inhibited by AII (A) and TPA (T) alone. Combined A + F and T + F dramatically ablated the F response. Cortisol levels (EIA) increased upon F treatment and were inhibited by A and T. Combination of treatments partially inhibited the F‐induced response. The protein‐coding region of marmoset and rhesus P450c17 cDNAs were then isolated from adrenals using RT‐PCR/TA cloning. Marmoset P450c17 shows one amino acid deletion but otherwise shares 90.6% and 91.4% homologies with the human and rhesus cDNA sequences, and 82.4% and 85% homologies with the human and rhesus predicted AA sequences, respectively. Since marmoset adrenocortical cells exhibit similar endocrine function to rhesus, impaired 17,20‐lyase activity in the adult marmoset adrenal may in part be due to differences in the primary sequence.