抑制pit-1基因表达对垂体生长激素腺瘤生物学行为影响的实验研究  

Effect of pituitary-specific transcription factor-1 antisence oligodeoxynucleotide on biological behavior of the growth hormone adenomas

作　　者：范月超[1] 张洪涛[1] 王雄伟[1] 张华楸[1] 舒凯[1] 万峰[1] 牛洪泉[1] 雷霆[1] 

机构地区：[1]华中科技大学同济医学院附属同济医院神经外科,武汉市430030

出　　处：《实用医学杂志》2005年第23期2597-2600,共4页The Journal of Practical Medicine

摘　　要：目的:研究垂体特异转录因子-1(pit-1)反义寡核苷酸(ASODN)对垂体生长激素(GH)腺瘤细胞增殖和细胞凋亡的影响,为以pit-1为靶点治疗GH腺瘤提供理论和实验依据。方法:设计合成硫代磷酸化反义寡核苷酸(PS-ASODN),经脂质体(lipofectin)包裹转染瘤细胞,每隔24h取样。采用四甲基偶氮唑蓝(MTT)法检测ASODN对细胞生长增殖的影响;用流式细胞仪(FCM)检测细胞凋亡和周期分布的改变,用免疫组化SABC法、逆转录-聚合酶链式反应(RT-PCR)法测定经ASODN作用后,pit-1蛋白和pit-1mRNA表达的变化。结果:ASODN组与正义组(SODN)及对照组相比较,48h、72h后细胞生长增殖受抑制(P<0.05),96h、120h时,差异有非常显著性(P<0.01);ASODN组对细胞凋亡有显著的促进作用,细胞多阻滞于G2/M期。其作用具有序列选择的特异性,在一定的时间范围内,呈时间依赖性。ASODN显著地抑制pit-1蛋白和pit-1mRNA的表达(均P<0.05)。结论:pit-1反义寡核苷酸能够有效地抑制GH腺瘤细胞的生长而促进其凋亡,具有较好的临床应用前景。Objective To explore the effect of pituitary-specific transcription factor （pit-1） antisence oligonucleotides（ASODN） on the proliferation and apoptosis of the growth hormone （GH） adenomas cells, and gain further insight about pit-1 as a target to the treatment of pituitary adenomas. Methods After being covered with lipofectin, the ASODN of telomerase was co-incubated with cultured GH adenomas cells. The cell survival rate was detected with MTT method. Cell apoptosis rate and the phase of cell cycle were determined by flow cytometer （FCM）. The changes of expression of pit-1 protein were detected by immunohistochemical technique, and the expression of pit-1 mRNA was assayed by RT-PCR. Results Compared with SODN and control groups, ASODN inhibited cell proliferation after being applied for 48 h, 72 h （ P 〈 0.05）, and markedly at 96 h, 120 h （ P 〈 0.01 ） . After 72 h, the growth of most transfected GH adenomas cells stopped at G2/M phase, and the number of apoptosis cells increased as time went on. The efficacy had the sequence selectivity and depended on time in a certain range. ASODN inhibited markedly the expression of pit-1 protein and pit-1 mRNA. Conclusion ASODN inhibited GH adenomas cells proliferation and promoted cells apoptosis, so it has a potential role in pituitary adenomas treatment.

关 键 词：垂体肿瘤 生长激素 腺瘤 垂体特异转录因子 寡核苷酸类 反义 

分 类 号：R736.4[医药卫生—肿瘤] R392.12[医药卫生—临床医学]