奥沙利铂对胃癌凋亡相关蛋白PDCD5和XIAP表达影响研究  

Effect of oxaliplatin chemotherapy on the expression of gastric cancer cell apoptosis related proteins PDCD5 and XIAP

作　　者：田景中[1] 李光云[1] 许建国[1] 史伟[1] 黄宝玉[1] 许戈良 TIAN Jing-zhong;LI Guang-yun;XU Jian-guo;SHI Wei;HUANG Bao-yu;XU Ge-liang(Department of Gastrointestinal Surgery, Bozhou People＇s Hospital, Bozkou 236804, P. R. China;Department of General Sugery ,Anhui Provincial Hospital, the First Affiliated Hospital of USTC , Hefei 230001 ,P. R. China)

机构地区：[1]亳州市人民医院胃肠外科,安徽亳州236804 [2]中国科学技术大学附属第一医院.安徽省立医院普外科,安徽合肥230001

出　　处：《中华肿瘤防治杂志》2018年第6期384-388,共5页Chinese Journal of Cancer Prevention and Treatment

基　　金：安徽省自然科学基金青年基金(1608085QH198)

摘　　要：目的 PDCD5和XIAP均为细胞凋亡关键分子,然而晚期胃癌奥沙利铂化疗与PDCD5和XIAP表达水平相关性尚不明确。本研究探讨奥沙利铂对胃癌组织细胞凋亡及凋亡诱导因子程序性细胞死亡蛋白-5(programmed cell death5,PDCD5)和凋亡抑制因子X-连锁凋亡抑制蛋白(X-linked inhibitor of apoptosis protein,XIAP)表达水平的影响。方法将活性良好的1×107个MGC-803人胃癌细胞接种于裸鼠背部皮下,制备裸鼠胃癌荷瘤模型,HE染色分析肿瘤组织病理形态学。根据实验目的,将荷瘤小鼠随机分为奥沙利铂组、生理盐水组和空白组,每组6只,接种胃癌细胞2周后,奥沙利铂组予以腹腔给药(25mg/kg),生理盐水组予以腹腔注射生理盐水(25mg/kg),空白组不予以特殊处理。每周1次,给药6次后处死动物,剥离肿瘤并组织匀浆,实时荧光定量PCR(real-time quantitative PCR,qPCR)检测各分组细胞凋亡诱导因子PDCD5和凋亡抑制因子XIAP基因表达,蛋白质印迹法评估各组PDCD5和XIAP蛋白表达,并对比分析各分组肿瘤组织质量以评估治疗效果,TUNEL染色检测肿瘤组织内细胞凋亡情况。实验数据采用单因素多样本方差分析。结果 MGC-803人胃癌细胞皮下接种裸鼠2周即可成瘤,成瘤大小约1.5cm×2.0cm。qPCR结果显示,与生理盐水组、空白组相比,奥沙利铂组显著上调细胞凋亡诱导因子PDCD5基因表达(F=49.452,P<0.001),降低凋亡抑制因子XIAP基因表达水平,F=12.467,P=0.001。蛋白质印迹法证实,与空白组或生理盐水组相比,奥沙利铂组高表达PDCD5蛋白(F=89.954,P<0.001),低表达XIAP蛋白,F=13.032,P=0.001。处死裸鼠后,对比各分组肿瘤组织重量,奥沙利铂组化疗能使裸鼠MGC-803人胃癌移植瘤体积显著缩小,F=9.828,P=0.002。TUNEL染色证实奥沙利铂组移植瘤内细胞凋亡水平最高,F=117.148,P<0.001。结论奥沙利铂化疗能显著增加小鼠胃癌组织内凋亡诱导因子PDCD5表达,同时也明显降低凋亡抑制因�OBJECTIVE Both PDCD5 and XIAP are key molecules of cell apoptosis,however,the correlation between oxaliplatin chemotherapy and PDCD5/XIAP is not clear in advanced gastric cancer.The aim of this study is to investigate the effects of oxaliplatin chemotherapy on gastric cancer cell apoptosis and the expression of apoptosis related proteins,apoptosis-inducing factor for Programmed Cell Death 5（PDCD5）and inhibitor of apoptosis protein,X-linked inhibitor of apoptosis protein（XIAP）in the xenografted tumor of nude mice.METHODS Nude mice were implanted with 1×10^7 high-activity MGC-803 human gastric cancer cells and the nude mice model of xenografted gastric cancer were built,then the tumor morphology was observed by HE staining.According to the experiment aim,all tumor-bearing mice were randomly divided into oxaliplatin group,normal saline group and blank group with 6 mice in each group.After 2 weeks of gastric cancer cells transplantation,oxaliplatin group was treated with 25 mg/kg oxaliplatin by intraperitoneal injection,normal saline group was treated with 25 mg/kg normal saline by intraperitoneal injection and the blank group was untreated.The drug injection was executed once a week.Nude mice were sacrificed six weeks later.The xenografted tumor was peeled to prepare tissue homogenate,and the gene expression of PDCD5 and XIAP were assayed by real-time quantitative PCR（qPCR）.The PDCD5 and XIAP protein expression was detected by western blot.Then,tumor weight for each group was analyzed to evaluate the treatment effect.Finally,the cell apoptosis of the transplanted tumor was evaluated by TUNEL staining.The experimental data were statistically analyzed by One-way analysis of variance.RESULTS After 2 weeks of MGC-803 gastric cancer cells transplantation in subcutaneous area of nude mice,tumor size was detected at about 1.5 cm×2.0 cm.The results of qPCR indicated that,compared with the blank group or normal saline group,oxaliplatin group significantly raised the gene expression of apoptosis inducing factor

关 键 词：奥沙利铂 胃癌 细胞凋亡 程序性细胞死亡蛋白-5 X-连锁凋亡抑制蛋白 

分 类 号：R735.2[医药卫生—肿瘤]