节律基因Period2在人卵巢上皮性癌组织中表达的意义及在裸鼠移植瘤组织中过度表达对移植瘤生长的作用  

Significance of circadian gene Period2 expression in epithelial ovarian cancer and the effect of gene overexpression on growth of ovarian cancer xenografts in nude mice

作　　者：王朝霞[1] 李莉[1] 

机构地区：[1]山西医科大学第一医院妇科,太原030001

出　　处：《中华妇产科杂志》2015年第6期446-451,共6页Chinese Journal of Obstetrics and Gynecology

基　　金：山西省科技攻关项目（20120313019-1）

摘　　要：目的探讨节律基因Period2在卵巢上皮性癌（卵巢癌）患者癌组织中表达的意义，并探讨Period2基因在裸鼠移植瘤组织中过度表达对移植瘤生长的作用及可能机制。方法选择2010年1月至2013年12月山西医科大学第一医院存档的卵巢癌石蜡标本22份（卵巢癌组），其中Ⅰ期8份、Ⅱ期8份、Ⅲ期6份，另选取同期卵巢良性上皮性肿瘤石蜡标本6份（良性肿瘤组），采用实时定量PCR技术和蛋白印迹（westernblot）法分别检测两组患者肿瘤组织中Period2mRNA和蛋白的表达。使用卵巢癌细胞株SKOV3细胞构建卵巢癌裸鼠移植瘤模型，实验分为3组，即重组质粒组、空质粒组、对照组，每组8只裸鼠；利用基因转染技术转染Period2基因，采用实时定量PCR技术检测转染后3组裸鼠移植瘤组织中Period2mRNA的表达，监测裸鼠移植瘤的生长情况，并计算其抑瘤率；采用实时定量PCR技术和westernblot法分别检测转染后3组裸鼠移植瘤组织中抗凋亡基因BRE、促凋亡基因TNFR1、抑癌基因NIXmRNA和蛋白的表达。结果（1）患者肿瘤组织中Period2mRNA的表达水平，卵巢癌组Ⅰ、Ⅱ、Ⅲ期分别为2．59±0．50、0．47±0．08、0．42±0．08，良性肿瘤组为6．59±1．05；患者肿瘤组织中Period2蛋白的表达水平，卵巢癌组Ⅰ、Ⅱ、Ⅲ期分别为0．835±0．087、0．412±0．035、0．199±0．031，良性肿瘤组为0．874±0．094。除良性肿瘤组Period2蛋白与Ⅰ期卵巢癌比较差异无统计学意义（p〉0．05）外，良性肿瘤组Period2mRNA和蛋白的表达水平均明显高于卵巢癌组Ⅰ、Ⅱ、Ⅲ期（P〈0．01）；且随着卵巢癌期别的升高，Period2mRNA和蛋白的表达水平明显降低（P〈0．05）。（2）转染后2周，重组质粒组裸鼠移植瘤组织中Period2mRNA的表达水平为6．11±0．56，明显高于空质粒组、对照组（分别为0．50±0．09、0．44±0．08；P〈0．01）；重组质粒组移�Objective To study the significance of circadian gene Period2 expression in epithelial ovarian cancer tissues and the effect of gene overexpression On the growth of ovarian cancer xenografts in nude mice. Methods Twenty-two cases of ovarian cancer paraffin specimens in the First Hospital of Shanxi Medical University （ovarian cancer group） were chosed during Jau. 2010 to Dec. 2013, including 8 cases of stageⅠ, 8 cases of stage Ⅱ, and 6 cases of stage Ⅲ, while 6 cases of benign ovarian epithelial tumor paraffin specimens were selected as control （benign tumor group）. Period2 gene were detected by real-time quantitative PCR and western blot methods in different stages of ovarian cancer tumor tissues. Established theovarian cancer xenografts in nude mice with ovarian cancer cell line SKOV3, and they weredivided into 3 groups （n=8）, including the recombinant plasmid group, empty plasmid group and control group. Using gene transfection technique to transfer Period2 gene into tumor tissues, tested the expression of Period2 mRNA in tumor tissues by real-time quantitative PCR after transfectiou into all nude mice, monitoredthetransplant tumor growth and calculating the tumor inhibition rate,detected the antiapoptotic gene BRE, apoptosis related tumor necrosis factor receptor （TNFR 1） andtumor suppressor gene NIX in tumor tissues by real-time PCR and western blot in different groups. Results （ 1 ） The expression level of Period2 mRNA in tumor tissues amongovarian cancer group stage Ⅰ, Ⅱand m were respectively 2.59±0.50, 0.47± 0.08 and 0.42±0.08, but benign tumor group was 6.59± 1.05. The expression level of Period2 protein in ovarian cancer group stage Ⅰ, Ⅱ and Ⅲ were respectively 0.835±0.087, 0.412±0.035 and 0.199±0.031, while benign tumor group was 0.874±0.094. The expression level of Period2 mRNA and protein in benign tumor group was higher than those in ovarian cancer group stage Ⅰ, Ⅱ or m （P〈0.01）. With ovarian cancer stage increased, the expression of Period2 mRNA

关 键 词：卵巢肿瘤 周期昼夜节律蛋白质类 肿瘤移植 受体 肿瘤坏死因子 I型 肿 瘤抑制蛋白质类 原癌基因蛋白质类 膜蛋白质类 

分 类 号：R737.31[医药卫生—肿瘤]