CHFR基因和PARP-1基因对裸鼠Raji淋巴瘤细胞增殖和凋亡调控的研究  

CHFR and PARP-1 regulate the proliferation and apoptosis of human Raji lymphoma xenografts in nude mice

作　　者：刘小丹[1] 宋亮[1] 管恩本[1] 潘华[1] 吕振梅[1] 刘建科[1] 徐雷[1] 宋爱琴[1] LIU Xiaodan;SONG Liang;GUAN Enben;PAN Hua;LV Zhenmei;LIU Jianke;XU Lei;SONG Aiqin(Department of Pediatrics,The Affiffiffiliated Hospital of Qingdao University,Qingdao 266000,China)

机构地区：[1]青岛大学附属医院儿童医学中心,青岛266000

出　　处：《中国小儿血液与肿瘤杂志》2023年第4期217-224,共8页Journal of China Pediatric Blood and Cancer

基　　金：山东省自然科学基金项目(Y2008C170);青岛市自然科学基金项目[2012-1-3-2-(5)-nsh]。

摘　　要：目的CHFR基因是一种抑癌基因,低表达或表达缺失可能促淋巴瘤细胞增殖,PARP-1基因参与其对淋巴瘤细胞增殖的调控,本研究旨在通过过表达CHFR基因及分别沉默CHFR、PARP-1基因,观察其对裸鼠移植瘤Raji细胞的影响,探讨CHFR基因和PARP-1基因调控裸鼠Raji淋巴瘤细胞的增殖和凋亡机制。方法在非霍奇金B细胞裸鼠Burkitt淋巴瘤移植瘤Raji细胞中,用5-氮杂-2′-脱氧胞苷(5-Aza-dC)过表达CHFR基因,及用慢病毒介导的shRNA分别沉默CHFR、PARP-1基因,测定各组裸鼠移植瘤的肿瘤的大小及重量;采用实时荧光定量PCR技术测定各组CHFR及PARP-1基因mRNA含量;应用MSP方法检测各组CHFR的甲基化状态;分别通过TUNEL试剂盒和免疫组化监测凋亡指数(AI)和微血管密度(MVD)。结果5-Aza-dC组的裸鼠在非霍奇金B细胞Burkitt淋巴瘤移植瘤Raji细胞能增强CHFR基因mRNA的表达,降低PARP-1基因mRNA的表达;在体内环境中,5-Aza-dC能够促进移植瘤Raji细胞的凋亡,抑制移植瘤的生长(P<0.05),这些结果与shRNA-CHFR组结果相反,与shRNA-PARP-1组结果一致。而且实验中可见5-Aza-dC组的裸鼠移植瘤的CHFR基因发生去甲基化。结论CHFR基因低表达促进肿瘤细胞增殖,PARP-1基因低表达促进肿瘤细胞凋亡,而5-Aza-dC可以去甲基化抑制肿瘤细胞增殖。Objective CHFR is a tumor suppressor gene,low or lack of CHFR expression may promote lymphoma cell proliferation.PARP-1 is involved in CHFR′s regulation of lymphoma cell proliferation.This study aimed to explore the effects of overexpression of CHFR and silencing of CHFR and PARP-1 on the proliferation and apoptosis of human Raji lymphoma xenografts in nude mice,so as to explore the mechanism of CHFR and PARP-1 regulating the proliferation and apoptosis of human Raji lymphoma xenografts in nude mice.Methods 5-Aza-2′-Deoxycytidine(5-Aza-dC)were used to overexpress CHFR,ShRNAs targeting CHFR and PARP-1 were transduced into Raji cells via lentivirus vector to silence CHFR and PARP-1 in human Raji lymphoma xenografts in nude mice.Tumor size and weight were determined using a xenograft model.Real-time PCR was carried out to determine the expression of CHFR and PARP-1.The methylation status of CHFR was measured through methylation-specific PCR(MSP)assay.The apoptotic index(AI)and micro-vessel density(MVD)were evaluated by terminal-deoxynucleotidyl transferase mediated nick end labeling(TUNEL)and immunohistochemistry.Results 5-Aza-dC-treatment promoted the mRNA expression of CHFR and decreased the mRNA expression of PARP-1 of human Raji lymphoma xenografts in nude mice.5-Aza-dC also accelerated Raji-cell apoptosis and restrained its growth in vivo(P<0.05).These results were contrary to those observed in the shRNA-CHFR group but consistent with those observed in the shRNA-PARP-1 group.Besides,treatment with 5-Aza-dC led to demethylation of CHFR in human Raji lymphoma xenografts in nude mice.Conclusions Our findings indicate that Low expression of CHFR gene promotes tumor cell proliferation,while low expression of PARP-1 gene m promotes tumor cell apoptosis.5-Aza-dC could lead to the demethylation of the CHFR promoter and suppress Raji cell growth.

关 键 词：B细胞淋巴瘤 RAJI细胞 CHFR基因 PARP-1基因 DNA甲基化 

分 类 号：R733.1[医药卫生—肿瘤]