摘要
背景:第10号染色体缺失的磷酸酶和张力蛋白同源基因(phosohataseandtensinhomolguedeletedon chromosome10,PTEN)的翻译变体(PTEN-Long)是新发现的一种肿瘤抑制基因,与多种肿瘤的发展密切相关。目的:探讨PTEN-Long对人脑胶质瘤U251细胞裸鼠异体移植瘤生长的抑制作用及相关分子机制。方法:(1)人脑胶质瘤细胞U251购自国家实验细胞资源共享服务平台北京总部,实验将未感染慢病毒的U251为对照组,感染LV-EGFP的U251为空载组,感染LV-PTEN-Long-EGFP的U251为过表达组,用荧光显微镜、Western blot检测转染效率;(2)实验将购自北京华阜康生物科技股份有限公司的BALB/c-nu雄性裸鼠随机分为3组,将3组细胞分别接种于裸鼠背部皮下,观察裸鼠一般状况及肿瘤生长趋势,分析各组裸鼠皮下肿瘤的体积及质量变化,免疫组织化学染色法检测肿瘤中PTEN-Long,p-Akt及NF-κB p65蛋白表达变化情况。结果与结论:(1)一般状况:PTEN-Long过表达后可抑制裸鼠皮下胶质瘤细胞的增殖,与对照组及空载组相比,过表达组裸鼠肿瘤体积及质量减少(P <0.01);(2)免疫组织化学染色显示:与对照组及空载组相比,过表达组PTEN-Long表达量增加(P<0.05),p-Akt及NF-κBp65表达量降低(P<0.05);(3)结果显示:PTEN-Long可抑制人脑胶质瘤U251细胞裸鼠异体移植瘤的生长,机制可能与下调PI3K-AKT-NF-κB信号通路有关。
BACKGROUND:PTEN-Long is a translational variant of phosohatase and tensin homolgue deleted on chromosome 10(PTEN),which is a newly discovered tumor suppressor gene and is closely related to the development of various tumors.OBJECTIVE:To investigate the inhibition effect of PTEN-Long on the growth of U251 cells(human glioma cells)in nude mice xenografts and related molecular mechanisms.METHODS:Human glioma cell U251 was purchased from the National Infrastructure of Cell Line Resource in Beijing.U251 not infected with lentivirus was used as control group.U251 infected with LV-EGFP was empty group.U251 infected with LV-PTEN-Long-EGFP was overexpressed group.Transfection efficiency was measured by fluorescence microscopy and western blot assay.BALB/c-nu male nude mice purchased from Beijing Huakang Biotechnology Co.,Ltd.were randomly divided into three groups,and the three groups of cells were inoculated subcutaneously into the back of nude mice.The general condition and tumor growth trend were observed.The volume and weight of subcutaneous tumors in nude mice were analyzed.The expression levels of PTEN-Long,p-Akt and NF-κB p65 protein in tumors were detected by immunohistochemical staining.RESULTS AND COUNCLUSION:Overexpression of PTEN-Long inhibited the proliferation of subcutaneous glioma cells in nude mice.Compared with the control group and the empty group,the overexpressed group was in poor condition,tumor volume and weight were decreased(P<0.01).Immunohistochemical staining showed that the expression of PTEN-Long(P<0.05)was increased,and the expression levels of p-Akt and NF-κB p65 were decreased(P<0.05).In summary,overexpression of PTEN-Long inhibits the growth of U251 cells in nude mice,and the mechanism may be related to down-regulation of PI3K-AKT-NF-κB signaling pathway.
作者
耿连婷
李春晖
单小松
郑亚北
曾昭穆
郑克彬
Geng Lianting;Li Chunhui;Shan Xiaosong;Zheng Yabei;Zeng Zhaomu;Zheng Kebin(Hebei University,Baoding 071000,Hebei Province,China;Department of Neurosurgery,Affiliated Hospital of Hebei University,Baoding 071000,Hebei Province,China)
出处
《中国组织工程研究》
CAS
北大核心
2019年第11期1743-1748,共6页
Chinese Journal of Tissue Engineering Research
基金
河北省自然科学基金资助项目(H2016201238)
项目负责人:郑克彬~~
