[1]赵　岗,杜　伟,魏新亭,等.胶质瘤组织中Bcl-2相关抗凋亡基因3表达及其对U87细胞增殖和侵袭能力的影响[J].新乡医学院学报,2018,35(10):843-848.[doi:10.7683/xxyxyxb.2018.10.001]
　ZHAO Gang,DU Wei,WEI Xin-ting,et al.Expression of Bcl-2-associated anti-apoptotic gene 3 in glioma tissues and its effect on proliferation and invasion of U87 cells[J].Journal of Xinxiang Medical University,2018,35(10):843-848.[doi:10.7683/xxyxyxb.2018.10.001]
点击复制

胶质瘤组织中Bcl-2相关抗凋亡基因3表达及其对U87细胞增殖和侵袭能力的影响
分享到：

《新乡医学院学报》[ISSN:1004-7239/CN:41-1186/R]

卷:
35
期数:
2018年10
页码:
843-848
栏目:
基础研究
出版日期:
2018-10-05

文章信息/Info

Title:
Expression of Bcl-2-associated anti-apoptotic gene 3 in glioma tissues and its effect on proliferation and invasion of U87 cells
作者:
赵　岗1; 杜　伟2; 魏新亭2; 王进忠1; 程振国1
（1.新乡市中心医院神经外科，河南　新乡　453000；2.郑州大学第一附属医院神经外科，河南　郑州　451000）
Author(s):
ZHAO Gang1; DU Wei2; WEI Xin-ting2; WANG Jin-zhong1; CHENG Zhen-guo1
（1.Department of Neurosurgery，Xinxiang Central Hospital，Xinxiang 453000，Henan Province，China；2.Department of Neurosurgery，the First Affiliated Hospital of Zhengzhou University，Zhengzhou 451000，Henan Province，China）
关键词:
胶质瘤; Bcl-2相关抗凋亡基因3; 细胞增殖; 细胞侵袭
Keywords:
glioma; Bcl-2-associated anti-apoptosis gene 3; cell proliferation; cell invasion
分类号:
R739.4
DOI:
10.7683/xxyxyxb.2018.10.001
文献标志码:
A
摘要:
目的　探讨胶质瘤组织中Bcl-2相关抗凋亡基因3（BAG3）表达及其对U87细胞增殖和侵袭能力的影响。方法　选取2011年4月至2017年4月于新乡市中心医院手术切除的胶质瘤组织及癌旁组织（距瘤旁边缘>2 cm）标本各73例，采用免疫组织化学法检测胶质瘤组织和癌旁组织中BAG3蛋白表达；培养人胶质瘤U87细胞，待细胞融合度达70%以上时随机分为siRNA-BAG3组、siRNA-对照序列组和空白对照组，siRNA-BAG3组细胞转染BAG3干扰序列，siRNA-对照序列组细胞转染阴性对照序列，空白对照组细胞不作任何处理，转染后继续培养48 h。采用实时荧光定量聚合酶链反应检测U87细胞中BAG3 mRNA表达，Western blot法检测U87细胞中BAG3蛋白表达，细胞计数试剂盒-8检测U87细胞增殖情况，克隆形成实验检测U87细胞克隆能力，流式细胞仪检测U87细胞凋亡情况，Transwell小室检测U87细胞侵袭能力。结果　胶质瘤组织中BAG3蛋白高表达率为73.97%（54/73），低表达率为26.03%（19/73）；癌旁组织中BAG3蛋白高表达率为36.99%（27/73），低表达率为63.01%（46/73）；胶质瘤组织中BAG3蛋白高表达率显著高于癌旁组织（χ2=20.215，P<0.05）。胶质瘤组织中BAG3蛋白表达与患者的年龄、性别、肿瘤位置、肿瘤直径及Karnofsky评分无关（P>0.05），而与肿瘤分级有关（P<0.05）。siRNA-BAG3组、siRNA-对照序列组和空白对照组U87细胞中BAG3 mRNA相对表达量分别为1.42±0.11、2.17±0.12和2.31±0.18；siRNA-BAG3组、siRNA-对照序列组和空白对照组U87细胞中BAG3蛋白相对表达量分别为0.20±0.12、0.84±0.13和0.92±0.12；siRNA-BAG3组U87细胞中BAG3 mRNA及蛋白相对表达量显著低于siRNA-对照序列组和空白对照组（P<0.05）；siRNA-对照序列组与空白对照组U87细胞中BAG3 mRNA及蛋白相对表达量比较差异无统计学意义（P>0.05）。培养24、48、72、96 h时，siRNA-BAG3组U87细胞增殖能力显著低于siRNA-对照序列组和空白对照组（P<0.05）；siRNA-对照序列组与空白对照组U87细胞增殖能力比较差异无统计学意义（P>0.05）。siRNA-BAG3组、siRNA-对照序列组和空白对照组U87细胞克隆形成数分别为30.76±6.19、81.32±3.55、85.07±3.05；siRNA-BAG3组U87细胞克隆形成数显著少于siRNA-对照序列组和空白对照组（P<0.05）；siRNA-对照序列组与空白对照组U87细胞克隆形成数比较差异无统计学意义（P>0.05）。siRNA-BAG3组、siRNA-对照序列组和空白对照组细胞凋亡率分别为（21.43±6.89）%、（5.50±2.95）%、（7.37±3.31）%；siRNA-BAG3组细胞凋亡率显著高于siRNA-对照序列组和空白对照组（P<0.05）；siRNA-对照序列组与空白对照组细胞凋亡率比较差异无统计学意义（P>0.05）。siRNA-BAG3组、siRNA-对照序列组和空白对照组侵袭细胞数分别为116.92±8.82、179.00±11.64、172.48±18.16，siRNA-BAG3组侵袭细胞数显著少于siRNA-对照序列组和空白对照组（P<0.05），siRNA-对照序列组与空白对照组侵袭细胞数比较差异无统计学意义（P>0.05）。结论　BAG3蛋白在胶质瘤组织中呈高表达，且与肿瘤恶性程度有关，特异性沉默人胶质瘤U87细胞中BAG3基因可有效抑制细胞增殖及侵袭能力。
Abstract:
Objective　To investigate the expression of Bcl-2-associated anti-apoptotic gene 3（BAG3） in glioma tissues and its effect on proliferation and invasion of U87 cells.Methods　The glioma tissues and adjacent tissues（from the edge of the tumor > 2 cm） from 73 patients with glioma were selected in Xinxiang Central Hospital from April 2011 to April 2017.The expressions of BAG3 protein in gliomas tissues and adjacent tissues were detected by immunohistochemistry.The human glioma U87 cells were cultured，and the U87 cells were randomly divided into siRNA-BAG3 group，siRNA-control sequence group and blank control group when the cell fusion degree was over 70%.The cells in the siRNA-BAG3 group were transfected with BAG3 interference sequence，and the cells in the siRNA-control sequence group were transfected with the negative control sequence，while the cells in the blank control group were not treated.The U87 cells were cultured for 48 hours after transfection.The expression of BAG3 mRNA in U87 cells was detected by real time fluorescence quantitative polymerase chain reaction.The expression of BAG3 protein in U87 cells was detected by Western blot.The proliferation of U87 cells was detected by cell counting kit-8.The cloning ability of U87 cells was detected by clone formation assay.The apoptosis of U87 cells was detected by flow cytometry.The invasion ability of U87 cells was detected by Transwell chamber.Results　The high expression rate of BAG3 protein in glioma tissues was 73.97%（54/73），and the low expression rate was 26.03%（19/73）.The high expression rate of BAG3 protein in adjacent tissues was 36.99%（27/73），and the low expression rate was 63.01%（46/73）.The high expression rate of BAG3 protein in glioma tissues was significantly higher than that in adjacent tissues（χ2=20.215，P<0.05）.The expression of BAG3 protein in glioma tissues was not correlated with age，gender，tumor location,tumor diameter and Karnofsky score（P>0.05）；but the expression of BAG3 protein in glioma tissues was correlated with tumor grade（P<0.05）.The relative expression levels of BAG3 mRNA in U87 cells of siRNA-BAG3 group，siRNA-control sequence group and blank control group were 1.42±0.11，2.17±0.12 and 2.31±0.18，respectively；the relative expression levels of BAG3 proteins in U87 cells of siRNA-BAG3 group，siRNA-control sequence group and blank control group were 0.20±0.12，0.84±0.13 and 0.92±0.12,respectively.The relative expression levels of BAG3 mRNA and protein in U87 cells in siRNA-BAG3 group were significantly lower than those in the siRNA-control sequence group and the blank control group（P<0.05）；there was no significant difference in relative expression level of BAG3 mRNA and protein between siRNA-control sequence group and the blank control group（P>0.05）.When cultured for 24，48，72 and 96 hours，the proliferation ability of U87 cells in siRNA-BAG3 group was significantly higher than that in the siRNA-control sequence group and the blank control group（P<0.05），but there was no significant difference in the proliferation ability of U87 cells between siRNA-control sequence group and blank control group（P>0.05）.The number of clone of U87 cells in siRNA-BAG3 group，siRNA-control sequence group and blank control group was 30.76±6.19，81.32±3.55 and 85.07±3.05,respectively.The number of clone of U87 cells in siRNA-BAG3 group was significantly less than that in the siRNA-control sequence group and the blank control group（P<0.05）.There was no significant difference in the number of clone of U87 cells between the siRNA-control sequence group and the blank control group（P>0.05）.The apoptosis rates of U87 cells in the siRNA-BAG3 group，siRNA-control sequence group and blank control group were （21.43±6.89）%，（5.50±2.95）%and （7.37±3.31）%，respectively.The apoptosis rate of U87 cells in the siRNA-BAG3 group was significant higher than that in the siRNA-control sequence group and the blank control group （P<0.05）.There was no significant difference in apoptosis rate of U87 cells between the siRNA-control sequence group and blank control group （P>0.05）.The number of invasive cells in the siRNA-BAG3 group，siRNA-control sequence group and blank control group was 116.92±8.82，179.00±11.64 and 172.48±18.16,respectively.The number of invasive cells in the siRNA-BAG3 group was significantly less than that in the siRNA-control sequence group and the blank control group （P<0.05）.There was no significant difference in the number of invasive cells between the siRNA-control sequence group and the blank control group（P>0.05）.Conclusion　BAG3 protein is highly expressed in glioma tissues，and it was associated with malignancy.Specific silencing of BAG3 gene in human glioma U87 cells can effectively inhibit cell proliferation and invasion.

参考文献/References:

［1］　MCGRANAHAN T，LI G，NAGPAL S.History and current state of immunotherapy in glioma and brain metastasis［J］.Ther Adv Med Oncol，2017，9（5）：347-368.
［2］　杨文涛，于喜贞，李正友，等.胶质瘤综合治疗方法研究进展［J］.新乡医学院学报，2016，33（1）：70-72，77.
［3］　WALSH K M，WIENCKE J K，LACHANCE D H，et al.Telomere maintenance and the etiology of adult glioma［J］.Neuro Oncol，2015，17（11）：1445-1452.
［4］　BEHL C.Breaking BAG：the co-chaperone BAG3 in health and disease［J］.Trends Pharmacol Sci，2016，37（8）：672-688.
［5］　SHERMAN M Y，GABAI V L.Hsp70 in cancer，back to the future［J］.Oncogene，2015，34（32）：4153-4161.
［6］　LOUIS D N，OHGAKI H，WIESTLER O D，et al.The 2007 WHO classification of tumours of the central nervous system［J］.Acta Neuropathol，2007，114（2）：97-109.
［7］　HU W，YANG Y，XI S，et al.Expression of CPEB4 in human glioma and its correlations with prognosis［J］.Medicine（Baltimore），2015，94（27）：e979.
［8］　BUNEVICIUS A，DELTUVA V P，TAMASAUSKAS A.Association of pre-operative depressive and anxiety symptoms with five-year survival of glioma and meningioma patients：a prospective cohort study［J］.Oncotarget，2017，8（34）：57543-57551.
［9］　赵建辉，胡世颉，李兵，等.胶质瘤临床诊疗研究进展［J］.中华神经外科疾病研究杂志，2017，16（4）：374-377.
［10］　单峤，王新军，王世勋，等.促红细胞生成素产生肝细胞受体A2在儿童脑胶质瘤中的表达及其意义［J］.中华实用儿科临床杂志，2014，29（5）：357-360.
［11］　LI M，HAN S，SHI X.In situ dendritic cell vaccination for the treatment of glioma and literature review［J］.Tumour Biol，2016，37（2）：1797-1801.
［12］　ZHU H，WU W，FU Y，et al.Overexpressed BAG3 is a potential therapeutic target in chronic lymphocytic leukemia［J］.Ann Hematol，2014，93（3）：425-435.
［13］　LIU B Q，ZHANG S，LI S，et al.BAG3 promotes stem cell-like phenotype in breast cancer by upregulation of CXCR4 via interaction with its transcript［J］.Cell Death Dis，2017，8（7）：e2933.
［14］　QIU S，SUN L，JIN Y，et al.Silencing of BAG3 promotes the sensitivity of ovarian cancer cells to cisplatin via inhibition of autophagy［J］.Oncol Rep，2017，38（1）：309-316.
［15］　KARPEL-MASSLER G，B M，SHU C，et al.TIC10/ONC201 synergizes with Bcl-2/Bcl-xL inhibition in glioblastoma by suppression of Mcl-1 and its binding partners in vitro and in vivo［J］.Oncotarget，2015，6（34）：36456-36471.

相似文献/References:

[1]王剑新,周文科,周国胜.靶向血管治疗脑胶质瘤[J].新乡医学院学报,2006,23(01):000.
[2]王永霞,王金祥.人脑胶质瘤中核转录因子-κB p65 和基质金属蛋白酶-9 的表达及临床意义[J].新乡医学院学报,2012,29(06):425.
[3]温　辉.G蛋白信号调节因子5动态表达与大鼠胶质瘤生长的关系[J].新乡医学院学报,2017,34(7):560.[doi:10.7683/xxyxyxb.2017.07.002]
　WEN Hui.Relationship between the dynamic expression of regulator of G-protein signaling 5 and the growth of glioma in rats[J].Journal of Xinxiang Medical University,2017,34(10):560.[doi:10.7683/xxyxyxb.2017.07.002]
[4]王怡春,黄毓杰,钟根深.分化抑制因子2对胶质瘤发生和发展的影响[J].新乡医学院学报,2022,39(1):088.[doi:10.7683/xxyxyxb.2022.01.019]
　WANG Yichun,HUANG Yujie,ZHONG Genshen.Effect of inhibitor of differentiation 2 on the occurrence and development of glioma[J].Journal of Xinxiang Medical University,2022,39(10):088.[doi:10.7683/xxyxyxb.2022.01.019]
[5]张玉松,孙玉学,韩亚迪.胶质瘤大鼠胶质瘤组织中果蝇zeste基因增强子的人类同源物2的表达及意义[J].新乡医学院学报,2022,39(4):308.[doi:10.7683/xxyxyxb.2022.04.002]
　ZHANG Yusong,SUN Yuxue,HAN Yadi.Expression and significance of enhancer of zeste homolog 2 in glioma tissue of rats with gliomas[J].Journal of Xinxiang Medical University,2022,39(10):308.[doi:10.7683/xxyxyxb.2022.04.002]
[6]杨文涛,于喜贞,李正友,等.胶质瘤综合治疗方法研究进展[J].新乡医学院学报,2016,33(1):070.[doi:10.7683/xxyxyxb.2016.01.021]
　YANG Wentao,YU Xizhen,LI Zhengyou,et al.Research progress of comprehensive treatment of glioma[J].Journal of Xinxiang Medical University,2016,33(10):070.[doi:10.7683/xxyxyxb.2016.01.021]

更新日期/Last Update: 2018-10-05