裸鼠体内人U343胶质瘤中Period2对胶质瘤p53的调控作用及意义  

Effect and Significance of which Period2 Regulated p53 in Human U343 Glioma in Vivo of Nude Mice

作　　者：王成泉 王珺[1,3] 尹磊[3] 夏鹤春[3] 

机构地区：[1]宁夏医科大学,银川750004 [2]宁夏医科大学颅脑疾病重点实验室,银川750004 [3]宁夏医科大学总医院,银川750004

出　　处：《宁夏医科大学学报》2015年第6期620-625,共6页Journal of Ningxia Medical University

基　　金：国家自然科学基金(81160313)

摘　　要：目的探讨裸鼠体内Period2(Per2)基因对人U343胶质瘤细胞p53的调控作用及意义。方法体外培养特异性人Per2基因shRNA慢病毒载体转染细胞(shRNA-Per2组)、非干扰慢病毒载体转染细胞(shRNAcontrol组)、空白对照U343胶质瘤细胞(空白对照组),分别接种于裸鼠颈后皮下,处于标准饲养环境成瘤。qRT-PCR、Western blot技术分别检测shRNA-Per2组、shRNA-control组、空白对照组中胶质瘤Per2、MDM2、p53、ATM、c-myc的mRNA和蛋白含量。结果 Per2低表达加快裸鼠体内胶质瘤细胞的成瘤及肿瘤的生长,shRNA-Per2组胶质瘤中Per2、p53、ATM的mRNA含量均低于shRNA-control组和空白对照组(P均<0.05);而MDM2与c-myc的mRNA水平均高于shRNA-control组和空白对照组(P均<0.05);各个基因的空白对照组与shRNA-control组中mRNA表达差异无统计学意义(P均>0.05)。Western blot结果表明各组裸鼠体内胶质瘤中5种蛋白的表达与上述mRNA呈现相同的趋势。shRNA-Per2组胶质瘤中Per2蛋白表达(0.541±0.0516)低于shRNA-control组(0.854±0.0947)和空白对照组(0.787±0.0913)(P均<0.05),p53蛋白表达(0.707±0.126)低于shRNA-control组(1.160±0.0717)和空白对照组(1.300±0.105)(P均<0.05),ATM表达(0.364±0.0309)低于shRNA-control组(0.742±0.190)和空白对照组(0.719±0.175)(P均<0.05),而shRNA-Per2组胶质瘤中MDM2蛋白表达(0.777±0.145)高于shRNA-control组(0.520±0.0634)和空白对照组(0.483±0.096)(P均<0.05),c-myc蛋白表达(0.522±0.0624)高于shRNA-control组(0.280±0.0652)和空白对照组(0.237±0.0623)(P均<0.05)。而各个基因在空白对照组与shRNA-control组中相对蛋白表达水平差异无统计学意义(P均>0.05)。结论在胶质瘤发生发展中Per2基因促进p53和ATM的表达,降低MDM2和c-myc的表达;Per2可视为今后治疗胶质瘤的潜在靶点。Objective To investigate the regulation of Period2 gene( Per2) on p53 in human U343 glioma in nude mice. Methods In vitro,specific Per2 gene shRNA lentivirus vector- transfected cells( shRNA-Per2),non- interfered lentivirus vector- transfected cells( shRNA- control) and blank treated p53 wild-type human U343 glioma cells( blank) were nourished. These cells inoculated in nude mice subcutaneously neck,keeping the standard environment until tumor formed. mRNA and protein levels of Period2,MDM2,p53,ATM,c- myc were detected using qRT- PCR and western blotting. Results Per2 knockdown accelerated tumor to form and grow up in vivo. The shRNA- Per2 group expressed significantly less mRNA of Per2p53,ATM than the shRNA- control group and the blank group did( P < 0. 05) while they expressed significantly more mRNA c- myc and MDM2 than the shRNA- control group and the blank group did( P < 0. 05),In addition,the protein expression of each group showed a similar trend in five kinds of proteins. The shRNA- Per2 group expressed significantly less Per2 protein( 0. 541 ± 0. 0516) than the shRNA- control group( 0. 854 ± 0. 0947) and the blank group( 0. 787 ± 0. 0913) did( P < 0. 05),while they expressed significantly less p53 protein( 0. 707 ± 0. 126) than the shRNA- control group( 1. 160 ± 0. 0717) and the blank group( 1. 300 ± 0. 105) did( P < 0. 05). The shRNA- Per2 group expressed significantly less ATM protein( 0. 364± 0. 0309) than the shRNA- control group( 0. 742 ± 0. 190) and the blank group( 0. 719 ± 0. 175)( P <0. 05) while they expressed significantly more MDM2 protein( 0. 777 ± 0. 145) than the shRNA- control group( 0. 520 ± 0. 0634) and the blank group( 0. 483 ± 0. 096) did( P < 0. 05) and they expressed significantly more c- myc protein( 0. 522 ± 0. 0624) than the shRNA- control group( 0. 280 ± 0. 0652) and the blank group( 0. 237 ± 0. 0623) did( P < 0. 05). Conclusion The low expression of Period2 gene promoted the expressions of MDM2,c- myc genes and inhibited the expressions of p53,ATM genes. Per2 can

关 键 词：Period2 人U343胶质瘤 MDM2 P53 

分 类 号：R739.4[医药卫生—肿瘤]