无功能垂体腺瘤中杀菌/渗透增强折叠家族蛋白B1基因的表达及其对肿瘤侵袭性的作用研究  

Expression and role of BPIFB1 in invasiveness of non-functional pituitary adenomas

作　　者：陈贤斌[1] 吴泽睿 卢江龙[1] 蔡霖[1] 苏志鹏[1] Chen Xianbin;Wu Zerui;Lu Jianglong;Cai Lin;Su Zhipeng(Department of Neurosurgery,the First Affiliated Hospital of Wetuhou Medical University,Wenzhou 325000,China)

机构地区：[1]温州医科大学附属第一医院神经外科,浙江325000

出　　处：《中华神经外科杂志》2021年第6期598-604,共7页Chinese Journal of Neurosurgery

基　　金：浙江省自然科学基金(LY17H160052,LY19C070002);温州市公益性科技计划项目(Y20180152,Y2020061)。

摘　　要：目的探讨无功能垂体腺瘤中杀菌/渗透增强折叠家族蛋白Bl(BPIFBl)基因的表达情况及其对垂体腺瘤增殖和侵袭性的影响;方法利用高通量基因表达(GEO)公共数据库垂体腺瘤芯片的数据,通过生物信息学分析方法筛选出侵袭性相关基因;然后选取5例侵袭性无功能垂体腺瘤和5例非侵袭性无功能垂体腺瘤的组织标本,通过实时荧光定量PCK和免疫组织化学染色检测BPIFBI基因和蛋白的表达情况。在垂体腺瘤GH3细胞株中通过siRNA敲减BPIFB1的表达,采用细胞活性和细胞划痕实验等检测肿瘤细胞的增殖和迁移能力;采用实时荧光定量PCR测定基质金属蛋白酶(MMP)-2和MMP-9 mRNA的表达水平;采用蛋白质免疫印迹技术检测上皮间质转化(EMT)相关蛋白的表达水平t结果生物信息学分析结果提示,侵袭性垂体腺瘤中BPIFB1的表达水平低于非侵袭性垂体腺瘤(log2FC=-5.29,P<0.01);免疫组织化学染色结果显示,侵袭性无功能垂体腺瘤中BPIFB1的表达水平(0.53±0.10)明显低于非侵袭性无功能垂体腺瘤(1.02±0.44)(P<0.05);实时荧光定量PCR结果显示,BIMFBl mRNA的表达水平与肿瘤KnSp分级呈负相关(r_(s)=-0.88,P<0.01)。细胞活性实验结果显示,BPIFB1敲减GH3细胞的细胞活性(2.98±0.14)高于正常GH3细胞(2.14±0.05)(P<0.01);细胞划痕实验结果显示,BPIFB1敲减GH3细胞的迁移率[(43.82±3.74)%]高于正常0113细胞[(17.53±2.05)%](P<0.01)。实时荧光定量PCR结果显示,BPIFBI敲减GH3细胞MMP-2和MMP-9 mRNA的表达水平均高于正常GH3细胞(均P<0.05);蛋白质免疫印迹实验显示,BPIFB1敲减GH3细胞中EMT相关蛋白的表达水平与正常GH3细胞比较差异均有统计学意义(均P<0.05)。结论侵袭性无功能垂体腺瘤中BPIFBI的表达水平降低,BPIFB1可能通过调控MMP的表达及EMT来抑制肿瘤的增殖和侵袭能力。Objective To explore the expression characteristics of BPIFBI in non-functional pituitary adenomas(NFPAs)and its effect on the ability of proliferation and invasiveness of pituitary adenomas.Methods The microarray data of pituitary adenoma from the public GEO(Gene Expression Omnibus)database and bioinformatic analysis were used to screen out the genes related to invasiveness of pituitary adenomas.The expression level of BPIFB1 was analyzed by RT-qPCR and IHC staining in the selected tissue specimens from 5 specimens of invasive tumors and 5 specimens of non-invasive tumors respectively.Small interfering RNAs(siRNA)were used to knockdown the expression of BPIFB1 in GH3 cells;the cell viability was tested by the CCK-8 experiment;the wound healing assay was used to analyze the migration ability of GH3 cells.RT-qPCR and Western blot analysis were used respectively to explore the mRNA expression levels of matrix metalloproteinases MMP-2 and MMP-9 and the expression of proteins associated with epithelial-mesenchymal transition(EMT).Results Bioinformatics analysis showed that the expression of BPIFB1 decreased in invasive pituitary adenomas compared with non-invasive adenomas(log2FC=-5.29,P<0.01).Immunohistochemical staining showed that the expression of BPIFB1 in invasive NFPAs(0.53±0.10,n=5)was significantly lower than that in non-invasive NFPAs(1.02±0.44,n=5;P<0.05).The expression of BPIFB1 mRNA was negatively correlated with Knosp grade of pituitary adenoma(r_(s)=-0.88,P<0.01).Cell viability assays showed that the cell viability of BPIFB1 knockdown GH3 cells(2.98±0.14)was higher than that of control GH3 cells(2.14±0.05;0.01).Cell scratch assays showed that the migration rate of BPIFB1 knockdown GH3 cells(43.82%±3.74%)was higher than that of control GH3 cells(17.53%±2.05%;P<0.01).The expression levels of MMP-2 and MMP-9 mRNA of BPIFB1 knockdown GH3 cells were higher than those of control GH3 cells(both P<0.05).The expression levels of EMT related proteins of BPIFB1 knockdown GH3 cells were different from those

关 键 词：垂体肿瘤 基因表达 肿瘤侵润 基质金属蛋白酶 杀菌/渗透增强折叠家族蛋白B1 

分 类 号：R736.4[医药卫生—肿瘤]