PHLPP2对小鼠Burkitt淋巴瘤移植瘤的影响  

Effect of PHLPP2 in the Burkitt lymphoma xenografted tumor in mice

作　　者：吴怡珅 朱雄鹏[1] 郑艳[1] 辛鹏亮[1] 刘生全 彭群艺 李纯团[1] WU Yishen;ZHU Xiongpeng;ZHENG Yan;XIN Pengliang;LIU Shengquan;PENG Qunyi;LI Chuntuan(Department of Hematology,The Quanzhou First Hospital Affiliated to Fujian Medical University,Quanzhou 362000,China)

机构地区：[1]福建医科大学附属泉州第一医院血液内科,福建泉州362000

出　　处：《山东第一医科大学（山东省医学科学院）学报》2024年第11期641-646,共6页Journal of Shandong First Medical University ＆ Shandong Academy of Medical Sciences

基　　金：泉州市科技计划项目(2018T007R)。

摘　　要：目的 研究PH结构域富含亮氨酸重复序列的蛋白磷酸酶2(PH domain leucine-rich repeat-containing protein phosphatase 2,PHLPP2)基因过表达对伯基特淋巴瘤(Burkitt lymohima,BL)移植瘤的影响及其对蛋白激酶B(protein kinase B,AKT)信号通路相关蛋白表达的影响。方法 在BL细胞系(CA46)中构建PHLPP2稳定过表达模型,并接种于重度免疫缺陷(NOD/ShiltJGpt-Prkdcem26Cd52112rgem26Cd22/Gpt,NCG)小鼠构建异种移植瘤模型。观察小鼠成瘤情况,测量肿瘤大小,绘制瘤体生长曲线,实验终点对比两组小鼠肿瘤大小及质量,评估PHLPP2对肿瘤生长的抑制效果。免疫组织化学染色检测PHLPP2、磷酸化蛋白激酶B(phosphorylated protein kinase B,p-AKT)蛋白表达情况,Western blot法检测PHLPP2、AKT、核糖体蛋白S6激酶β1 (ribosomal protein S6 kinase beta-1,S6K1)蛋白表达水平及AKT、S6K1蛋白磷酸化水平。结果 成功构建NCG小鼠BL动物模型:10只小鼠皮下均有肿瘤形成,成瘤率100%。PHLPP2基因对小鼠移植瘤的影响:对照组与实验组小鼠皮下移植瘤成瘤时间分别为(10.60±1.95)、(15.00±1.58)d;与对照组(2.00±1.01) mm3相比,实验组(0.63±0.18)mm3肿瘤体积明显减少;对照组肿瘤质量为(0.83±0.33)g,实验组为(0.31±0.03)g,差异均有统计学意义(P均<0.05)。Western blot及免疫组化结果:Western blot检测到实验组PHLPP2表达水平高于对照组,实验组p-AKT表达水平低于对照组,差异均有统计学意义(P均<0.05);免疫组化结果显示,实验组及对照组中PHLPP2蛋白及p-AKT蛋白在细胞膜、细胞质中表达,其中实验组PHLPP2蛋白呈强阳性,p-AKT蛋白呈弱阳性,对照组PHLPP2蛋白呈弱阳性,p-AKT呈强阳性,差异有统计学意义(P<0.05)。结论 PHLPP2在BL发生发展过程中发挥着重要的抑制作用,其抑制作用可能通过抑制AKT信号通路相关下游蛋白的磷酸化水平而实现。Objective:By studying the effect of PHLPP2 gene over-expression on Burkitt lymphoma grafts and its effect on AKT signaling pathway related protein expression,the mechanism of action of PHLPP2 on Burkitt lymphoma is further explored,providing experimental basis for the development of gene targeted therapy.Methods:The stable over-expression model of PHLPP2 is established in Burkitt lymphoma cell line(CA46)and inoculated into NCG mice to construct Xenograft tumor model.Tumor formation in mice is observed,tumor size is measured,and tumor growth curve is drawn.At the end of the experiment,the mice are euthanized to collect the samples of Xenografted tumor.The tumor volume and weight of the two groups are compared to evaluate the inhibitory effect of PHLPP2 on tumor growth.The expression of PHLPP2,p-AKT is detected by immunohistochemistry staining.and the expression levels of PHLPP2,AKT and S6K1 protein and the phosphorylation levels of AKT and S6K1 protein are detected by Western blot.Results:1.The NCG mouse model of Burkitt lymphom Xenograft model is successfully established:subcutaneous tumor formation is observed in all 10 mice,the tumor-forming rate is 100%.2.Effect of PHLPP2 gene on transplanted tumor in mice:the time of subcutaneous tumor formation in control group and treatment group are(10.60±1.95)and(15.00±1.58)respectively.Compared with the control group(2.00±1.01)mm3,the tumor volume of the treatment group(0.63±0.18)mm3 is significantly reduced(P<0.05).The average tumors weight in control group and treatment group are(0.83±0.33)g and(0.31±0.03)g(P<0.05).3.Western blot and immunohistochemical results:the results of Western blot indicate that the expression of PHLPP2 in the treatment group is significantly higher than that in the control group,while the expression of p-AKT and p-S6K1 in the treatment group is decreased comparing with the control group(P<0.05).Immunohistochemical results show that PHLPP2 protein and p-AKT protein are expressed in the cell membrane and cytoplasm in the experimental group

关 键 词：PH结构域富含亮氨酸重复序列的蛋白磷酸酶2 BURKITT淋巴瘤 NCG小鼠 移植瘤 

分 类 号：R733.1[医药卫生—肿瘤]