Adenovirus vectors differentially modulate proliferation of pituitary lactotrophs in primary culture in a mitogen and infection time-dependent manner.
Adenovirus vectors differentially modulate proliferation of pituitary lactotrophs in primary culture in a mitogen and infection time-dependent manner.
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腺病毒载体以丝裂原和感染时间依赖性方式差异调节原代培养物中垂体催乳素细胞的增殖。
DOI：
10.1677/joe-08-0102
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发表时间：
2008
期刊：
影响因子：
--
通讯作者：
J. Arita
中科院分区：
文献类型：
--
作者：
Zhenhuan Wang;Tetsuo Mitsui;Maho Ishida;J. Arita
腺病毒是一种功能强大、应用广泛的基因转移载体。然而，它们的应用限制并没有得到很好的描述。我们使用原代培养的大鼠垂体促乳素细胞作为模型，研究腺病毒载体感染如何调节丝裂原诱导的增殖和丝裂原信号通路的活性。携带β-半乳糖苷酶基因的腺病毒载体(Betagal)可提高基础增殖水平，并阻断胎牛血清(FBS)诱导的哺乳细胞增殖，但不影响Forsklin、IGF-I和溴隐亭诱导的增殖变化。表达β-雌二醇腺病毒不改变17β-雌二醇(E(2))在IGF-I存在下的抑制作用，但可阻断E(2)在葡聚糖包被的炭条血清或Forsklin存在下的刺激作用。表达NO蛋白的腺病毒不能阻断FBS诱导的增殖，但能有效地调节基础增殖水平和E(2)的刺激作用。基础增殖水平的升高和对FBS诱导的增殖的阻断是一过性的，并在感染后5d消失，而Forsklin对E(2)的刺激作用持续阻断。腺病毒感染使磷酸化的cAMP反应元件结合蛋白(PCREB)和ERK1/2的基础蛋白水平升高，pCREB免疫反应性乳酸菌比例增加。腺病毒还以一种基因特异性的方式改变了雌激素诱导的几个雌激素反应基因的mRNA表达。结果表明，腺病毒载体对不同有丝分裂原的乳酸菌增殖有不同程度的干扰作用。我们的结果提示，在正常细胞的原代培养中进行腺病毒基因转移时，必须考虑与所转移的基因无关的腺病毒的影响。
Adenoviruses are powerful, widely utilized vectors for gene transfer. Limitations to their application, however, have not been well described. We used rat pituitary lactotrophs in primary culture as a model for studying how adenovirus vector infection modulates mitogen-induced proliferation and the activities of mitogen signaling pathways. Infection with adenovirus vectors expressing beta-galactosidase (betagal) raised basal proliferative levels and blocked fetal bovine serum (FBS)-induced proliferation of lactotrophs, but did not influence the changes in proliferation induced by forskolin, IGF-I, and bromocriptine. The betagal-expressing adenoviruses did not alter the inhibitory action of 17beta-estradiol (E(2)) in the presence of IGF-I; however, they blocked the stimulatory action of E(2) in the presence of dextran-coated charcoal-striped serum or forskolin. An adenovirus expressing no protein failed to block FBS-induced proliferation, but was effective in modulating basal proliferative levels and the stimulatory actions of E(2). The increased basal proliferative level and the blockade of FBS-induced proliferation were transient, and lost 5 days after infection while the blockade of the stimulatory action of E(2) in the presence of forskolin persisted. Adenovirus infection raised basal protein levels of the phosphorylated forms of cAMP response element-binding protein (pCREB) and ERK1/2 and increased the proportion of pCREB-immunoreactive lactotrophs. Adenoviruses also altered estrogen-induced responses in mRNA expression of several estrogen-responsive genes in a gene-specific manner. The results demonstrate that an adenovirus vector differentially interferes with lactotroph proliferation in response to various mitogens. Our results suggest that the effects of the adenovirus that are independent of the genes transferred must be considered when performing adenoviral gene transfer in the primary cultures of normal cells.
影响因子：
20.3
作者：
N. Ben-Jonathan;R. Hnasko
通讯作者：
N. Ben-Jonathan;R. Hnasko
影响因子：
10.5
作者：
HSIAO, KM;MCMAHON, SL;FARNHAM, PJ
通讯作者：
FARNHAM, PJ
正常垂体促性腺激素基因转移的高效方法：腺病毒介导的 G 蛋白偶联受体激酶 2 表达抑制黄体生成素分泌。
影响因子：
4.8
作者：
Neill,JD;Musgrove,LC;Duck,LW;Sellers,JC
通讯作者：
Sellers,JC