GDNF重塑BV2细胞对胶质瘤相关因子的分泌能力  

GDNF remodels BV2 cells to secrete glioma-related factors

作　　者：李赛赛 王潇雨 高擎 荣玉如 张宝乐 LI Saisai;WANG Xiaoyu;GAO Qing;RONG Yuru;ZHANG Baole(Department of Cell Biology and Neurobiology,Xuzhou Key Laboratory of Neurobiology,Xuzhou Medical University,Xuzhou,Jiangsu 221004,China;Department of Clinical Medicine,Xuzhou Medical University,Xuzhou,Jiangsu 221002)

机构地区：[1]徐州医科大学神经生物学重点实验室,江苏徐州221004 [2]徐州医科大学临床医学系,江苏徐州221002

出　　处：《徐州医科大学学报》2020年第10期735-740,共6页Journal of Xuzhou Medical University

基　　金：国家自然科学基金项目(81602464);江苏省“六大人才高峰”项目(SWYY-088);江苏省“青蓝工程”优秀青年骨干教师项目(苏教办师[2017]5号);国家级大学生创新创业训练计划(201810313012);江苏省高等学校大学生实践创新训练计划(201810313011Z);基础医学国家级实验教学示范中心(徐州医科大学)资助项目;徐州市推动科技创新项目(KC20145)。

摘　　要：目的探讨胶质细胞系源性神经营养因子(GDNF)对小鼠BV2小胶质细胞分泌肿瘤相关细胞因子的影响。方法采用0、50、100、200和500μg/L GDNF分别处理BV2细胞12、24、36和48 h。实时PCR检测各组细胞中肿瘤抑制因子肿瘤坏死因子α(TNF-α)、干扰素β(IFN-β)和白细胞介素10(IL-10)和肿瘤促进因子IL-1β、转化生长因子β(TGF-β)、肝细胞生长因子(HGF)和基质金属蛋白酶9(MMP9)的mRNA水平。ELISA法检测TNF-α、IFN-β和IL-1β的蛋白分泌水平。利用GDNF和GFR-α1的中和抗体处理BV2细胞,并采用ELISA检测IL-1β蛋白的分泌量。结果GDNF显著抑制TNF-α和IFN-β的mRNA表达,同时显著升高IL-1β、TGF-β和MMP9的mRNA表达(P<0.05)。TNF-α、IFN-β和IL-1β的蛋白分泌水平与mRNA结果一致。GDNF和GFR-α1的中和抗体均显著抑制了IL-1β蛋白的分泌,且中和GFR-α1显著抑制了GDNF诱导的IL-1β蛋白的分泌(P<0.05)。结论GDNF处理的小胶质细胞低分泌肿瘤抑制因子、高分泌肿瘤促进因子,阻断GFR-α1能够抑制GDNF诱导的IL-1β蛋白分泌,提示GDNF可能经GFR-α1介导的信号通路重塑利于胶质瘤进展的微环境。Objective To investigate the effects of glial cell line-derived neurotrophic factor(GDNF)on mouse BV2 microglia cells to secrete tumor-related factors.Methods BV2 cells were treated with 0,50,100,200 and 500μg/L GDNF for 12,24,36 and 48 h.Real-time PCR was used to detect the mRNA levels of tumor suppressor factors tumor necrosis factorα(TNF-α),interferonβ(IFN-β)and interleukin 10(IL-10)and tumor promoting factors IL-1β,transforming growth factorβ(TGF-β),hepatocyte growth factor(HGF)and matrix metalloproteinase 9(MMP9).The neutralizing antibody of GDNF or GFR-α1 was used to treat BV2 cells,and the secretion of IL-1βprotein was detected by ELISA.Results GDNF significantly reduced the levels of TNF-αand IFN-βmRNA,while significantly increased the levels of IL-1β,TGF-β,MMP9 and IL-10 mRNA in BV2 cells(P<0.05).Moreover,the secretion of TNF-α,IFN-βand IL-1βprotein was consistent with their mRNA expression.Both GDNF and GFR-α1 neutralizing antibodies significantly inhibited the secretion of IL-1βprotein,and neutralizing GFR-α1 significantly inhibited the secretion of IL-1βprotein induced by GDNF(P<0.05).Conclusions GDNF-treated BV2 microglia cells secret low levels of tumor suppressor factors but high levels of tumor promoting factors,and GFR-α1 blockage inhibits GDNF-induced IL-1βprotein secretion,which indicates that GDNF may remold the microenvironment that is conducive to the progression of glioma through GFR-α1 mediated signaling pathway.

关 键 词：胶质细胞系源性神经营养因子 小胶质细胞 细胞因子 胶质瘤 肿瘤微环境 

分 类 号：R739.41[医药卫生—肿瘤]