Gadd45β及细胞周期调控相关基因在泡型包虫病患者肝脏中的表达及意义  

Gadd45β and regulation of the expression of cell cycle-related genes in patients with a alveolar echinococcosis and their significance

作　　者：杨舒婷[1,2] 张传山[2] 李亮[2] 毕晓娟[2] 林仁勇[2] 温浩[1,2] 

机构地区：[1]新疆医科大学第一附属医院临床研究院,新疆重大疾病医学重点实验室-省部共建国家重点实验室培育基地,新疆包虫病基础医学重点实验室,新疆乌鲁木齐830054 [2]新疆医科大学基础医学院生物学教研室,新疆乌鲁木齐830011

出　　处：《中国病原生物学杂志》2016年第12期1089-1093,共5页Journal of Pathogen Biology

基　　金：新疆维吾尔自治区自然科学基金青年基金项目(No.2014211C069)

摘　　要：目的研究肝泡型包虫病患者肝脏细胞Gadd45β(growth arrest and DNA damage gene 45β)及细胞周期调控相关基因的表达及意义。方法采集24例肝泡型包虫病患者肝脏标本,分为病灶旁组织(Close)和远端组织(Distance)。用重组TGF-β1(1ng/ml)及泡球蚴匀浆蛋白(1mg/ml)刺激体外培养的HL-7702人肝细胞,分别于30min、1h、6h、12h、24h、48h收集细胞。患者肝脏标本及刺激后收集的细胞均采用Trizol法提取RNA,反转录cDNA,通过实时荧光定量PCR检测目的基因表达并进行分析。结果近旁组织PCNA,TGF-β1,cyclinA1,cyclinE基因相对表达量分别为1.934±1.337、1.155±0.5138、1.367±0.8087、1.384±1.116、1.580±1.005、1.899±1.173和1.478±1.189,显著高于远端组织(1.00±0.00)且差异有统计学意义(P<0.05),而Gadd45β,cyclinB1,cyclinD差异无统计学意义(P>0.05)。用重组TGF-β1(1ng/ml)及泡球蚴匀浆蛋白(1mg/ml)刺激体外培养的HL-7702人肝细胞,引起Gadd45β,cyclinA1,cyclinE表达上调。结论泡球蚴匀浆蛋白刺激,可引起肝细胞Gadd45β及细胞周期相关蛋白表达上调,参与细胞抗凋亡过程。泡球蚴感染晚期,肝脏中仍存在细胞周期素(cyclins)参与的细胞增殖,但同时TGF-β1高表达,并通过Smads信号通路促进肝纤维化相关蛋白表达,促成肝脏泡型包虫病晚期病理变化。Objective To examine growth arrest- and DNA damage-inducible gene 4513 （Gadd45β） and regulation of the expression of cell cycle-related genes in patients with alveolar echinococeosis （AE） and their significance. Methods Liver tissue specimens were collected from 24 patients with AE, and tissue was divided into tissue adjacent to lesions （Near- by） and tissue distant from lesions （Distant）. HL-7702 liver, cells were cultured in vivo and stimulated with recombinant TGF-β1 （1 ng/ml） cell factor orEchinococcusmultilocularis proteins （1mg/ml） for 30 min, 1h, 6h, 12h, 24h, and 48h. mRNA was extracted from patient tissue and cell specimens and then reverse transcribed into cDNA. The expression of Gadd45β and cell cycle-related genes was analyzed using quantitative real-time PCR （qRT-PCR）. Results The rela- tive level of PCNA expression in nearby tissue was 1.934±1.337, the relative level of TGFq31 expression was 1.155±0.5138, the relative level of cyclinA1 expression was 1.367±0.8087, the relative level of cyclinE was 1.384±1.116, the relative level of Gadd45β expression was 1. 580±1. 005, the relative level of cyclinB1 expression was 1. 899±1. 178, and the relative level of expression was eyclinD was 1. 478±1. 189. Levels of PCNA, TGF-β1, cyclinA1, and cyclinE expression were significantly （P〈0.05） higher than those in distal liver tissue （1.00±0.00） while levels of Gadd45β, cyclinB1, and cyclinD did not differ significantly （P〉0.05）. In addition, stimulation with the recombinant cytokine TGF-β1 （1ng/ml） or E. multilocularis proteins （1 mg/ml） for 30 min, 1 h, 6 h, 12 h, 24 h, and 48 h up regulated the ex- pression of Gadd45β, cyclinA, and cyclinE mRNA in human HL-7702 liver cells. Conclusion Liver cells were stimulated with E. multilocularis proteins, causing up-regulation of the expression of Gadd45β and cell cycle-related proteins,and these molecules were involved in countering apoptosis. In the late stages of infection, cyelins induced cell prolif

关 键 词：泡型包虫病 GADD45Β 细胞周期 泡球蚴 

分 类 号：R532.32[医药卫生—内科学]