PRDM2和c-Myc在垂体生长激素腺瘤发生发展中作用的实验研究  

Experimental study of the roles of PRDM2 and c-Myc in the tumorigenesis and development of pituitary somatotroph adenomas

作　　者：张树恒 陈一元[1] 李振业 张亚卓[1] 高华[1] Zhang Shuheng;Chen Yiyuan;Li Zhenye;Zhang Yazhuo;Gao Hua(Beijing Neurosurgical Institute,Capital Medical University,Beijing 100070,China;Department of Neurosurgery,Anshan Central Hospital,Anshan 114001,China)

机构地区：[1]首都医科大学,北京市神经外科研究所,100070 [2]鞍山市中心医院神经外科,辽宁114001

出　　处：《中华神经外科杂志》2020年第8期841-846,共6页Chinese Journal of Neurosurgery

基　　金：国家自然科学基金(81601205);北京市卫计委高层次人才(2015-3-040)。

摘　　要：目的探讨视网膜母细胞瘤相互作用锌指蛋白1(PRDM2)和c-Myc在垂体生长激素腺瘤发生、发展中的作用。方法垂体腺瘤标本来自2013年7月至2016年12月首都医科大学附属北京天坛医院神经外科行手术治疗的70例垂体生长激素腺瘤患者,3例正常垂体组织标本来自遗体自愿捐献者。通过免疫组织化学染色方法检测肿瘤和正常垂体组织中PRDM2和c-Myc蛋白的水平。探讨肿瘤组织中PRDM2与c-Myc蛋白水平之间的关系,以及PRDM2和c-Myc蛋白与肿瘤侵袭能力、肿瘤体积的关系。体外培养大鼠垂体腺瘤GH3细胞株,设立对照组(空白细胞)、空载体组(转染空载体)和PRDM2组(转染PRDM2质粒)。通过细胞增殖实验检测PRDM2对细胞活力的影响;采用流式细胞术观察PRDM2对细胞凋亡的影响;行Transwell小室实验观察PRDM2对细胞侵袭能力的影响;采用蛋白质免疫印迹实验检测PRDM2对GH3细胞c-Myc表达水平的影响。结果根据70例肿瘤标本的免疫组织化学染色评分中位数(PRDM2:40分,c-Myc:170分)分组,PRDM2低评分组(<40分)较PRDM2高评分组(≥40分)有更高的侵袭机会(19/35对比9/35,P=0.015)和更大的肿瘤体积[(17.8±4.5)cm3对比(5.2±0.9)cm3,P<0.01]。c-Myc高评分组(≥170分)较c-Myc低评分组(<170分)有更高的侵袭机会(20/35对比8/35,P=0.003)和更大的肿瘤体积[(15.5±3.5)cm3对比(6.6±2.0)cm3,P=0.027]。70例肿瘤标本中的PRDM2与c-Myc水平呈负相关(r=-0.407,P=0.004)。GH3细胞转染24、48、72 h后,PRDM2组的细胞活力分别为对照组的(82.2±6.3)%、(68.5±5.2)%、(54.7±3.6)%(均P<0.05);而空载体组与对照组的差异均无统计学意义(均P>0.05)。流式细胞术检测结果显示,转染24 h后,PRDM2组膜联蛋白V(AnnexinⅤ)染色阳性细胞和碘化丙啶(PI)染色阳性细胞所占的比率分别为(14.66±3.72)%和(6.21±2.17)%,均高于对照组[分别为(3.35±1.03)%和(1.34±0.48)%,均P<0.05];而空载体组AnnexinⅤ阳性细胞和PI阳性细胞所占比Objective To explore the roles of PR domain zinc finger protein 2(PRDM2)and c-Myc in the tumorigenesis and development of somatotroph adenomas at the tissue and cellular levels.Methods Seventy somatotroph adenomas specimens were obtained from patients with somatotroph adenomas who underwent tumor resection at Department of Neurosurgery,Beijing Tiantan Hospital,Capital Medical University from July 2013 to December 2016.Three normal pituitary gland specimens were from donators of cadaver.The protein levels of PRDM2 and c-Myc were measured by immunochemistry.We further explored the relationship between PRDM2 and c-Myc protein levels in tumor tissue,as well as the relationship between PRDM2 and c-Myc proteins and tumor invasion and tumor volume.Somatotroph adenomas cell line GH3 cells were cultured in vitro and further divided into control group(blank cells),vector group(transfected with vectors only)and PRDM2 group(transfected with PRDM2 plasmid).Effect of PRDM2 on cell viability was detected by cell proliferation experiment,effect of PRDM2 on apoptosis was observed by flow cytometry,effect of PRDM2 on cell invasion ability was observed in the Transwell chamber experiment,and the effect of PRDM2 on the expression level of c-Myc in GH3 cells was detected by Western blot experiment.Results According to the median immunohistochemistry score of 70 tumor specimens(PRDM2:40,c-Myc:170),the PRDM2 low-score group(<40)had a higher chance of invasion(19/35 vs.9/35,P=0.015)and larger tumor volume(17.8±4.5 cm3vs.5.2±0.9 cm3,P<0.01)than the PRDM2 high-score group(≥40).The c-Myc high-score group(≥170)had a higher chance of invasion(20/35 vs.8/35,P=0.003)and larger tumor volume(15.5±3.5 cm3vs.6.6±2.0 cm3,P=0.027)than the c-Myc low-score group(<170).PRDM2 and c-Myc levels in 70 tumor specimens were negatively correlated(r=-0.407,P=0.004).After 24 hours,48 hours and 72 hours of GH3 cell transfection,the cell vitality of PRDM2 group was(82.2±6.3)%,(68.5±5.2)%and(54.7±3.6)%of that of control group respectively(all P<0.05),an

关 键 词：分泌生长激素的脑垂体腺瘤 原癌基因蛋白质C-MYC 视网膜母细胞瘤相互作用锌指蛋白1(PRDM2) 细胞增殖 肿瘤侵润 

分 类 号：R736.4[医药卫生—肿瘤]