腺病毒介导酪氨酸羟化酶基因治疗垂体泌乳素瘤实验研究

Prolactinomas are responsive to dopamine and its agonist, which bind to dopamine receptors on prolactinoma cells, whereby they not only decrease prolactin secretion, but also decrease adenoma size. Tyrosine hydroxylase (TH) is the rate-limiting enzyme in dopamine synthesis. This project is an experimental study of gene therapy of rat prolactinomas mediated by adenoviral vectors with a gene encoding rat tyrosine hydroxylase. Methods Recombinant replication-deficient adenovirus named Advth is constructed by traditional homologous recombination in 293 cells. Recombinant replication-deficient adenovirus named Ad-GFP-TH with rat TH-cDNA and control adenovirus named Ad-GFP were constructed by homologous recombination in bacterial cells. Fluorescopy, PCR, immunohistochemistry and dopamine detest are used to evaluate the recombinant adenovius. The rat pituitary prolactinoma cell line MMQ and oestrogen-induced Sprague-Dawley（SD）rat pituitary prolactinoma models are chosen as the target to study the effect of gene therapy on their growth and prolactin secretion mediated by Ad-GFP-TH. Results Recombinant Advth, Ad-GFP-TH and Ad-GFP are successfully reconstructed. Transfection of MMQ cells with Ad-GFP-TH in vitro not only restrains their growth but also decreases their PRL secretion significantly. Transfection of oestrogen-induced Sprague-Dawley（SD）rat pituitary prolactinoma with Ad-GFP-TH in vivo significantly decreases their PRL secretion, but have no influence on their growth. Conclusions Gene therapy of rat prolactinomas mediated by adenovirus with rat tyrosine hydroxylase gene has significant therapeutical effects. Gene therapy maybe a potential treatment for pituitary adenoma.

垂体腺瘤是最常见的颅内肿瘤之一，经手术和放疗常达不到生物学治愈。我们以大鼠垂体泌乳素（PRL）腺瘤为模型，研究基因治疗垂体腺瘤的可能性。本研究是基于多巴胺促效剂可允勾固逑倭鏊跣『头置赑RL降低，酪氨酸羟化酶（TH）是多巴胺合成的限速酶，我们拟将携带大鼠TH基因的腺病毒载体转染体外培养的大鼠垂体PRL腺瘤细胞和导入大鼠PRL腺瘤模型的垂体瘤组织，通过TH核酸杂交、免疫组化和PRL放免分析，观察本方法