摘要
目的探讨基质Gla蛋白(MGP)对髓母细胞瘤(MB)细胞增殖、凋亡的影响,初步明确MGP在儿童MB发病中的作用和机制。方法通过分析比较基因表达综合数据库筛选出待研究基因MGP。收集2018年1月至2021年12月郑州大学第三附属医院手术切除的14例MB患者的癌组织及配对癌旁正常组织,采用反转录-聚合酶链反应及Western blot法检测癌组织和癌旁正常组织中MGP蛋白相对表达量。培养MB细胞Daoy和D341,待细胞生长至融合度达到30%~50%时分为Doay空白组、Doay小干扰RNA(siR)-MGP组和D341空白组、D341 siR-MGP组。Doay siR-MGP组和D341 siR-MGP组细胞均转染siR-MGP,Doay空白组和D341空白组细胞不进行转染。分别于转染24、48、72 h,采用细胞计数试剂盒-8法检测4组细胞的增殖能力;转染48 h后采用流式细胞术检测4组细胞的凋亡率。结果MB癌组织中MGP蛋白的相对表达量显著高于癌旁正常组织(P<0.05)。转染24、48、72 h时,Doay空白组细胞增殖能力显著高于Daoy siR-MGP组,D341空白组细胞增殖能力显著高于D341 siR-MGP组(P<0.05)。Doay siR-MGP组细胞的凋亡率显著高于Doay空白组,D341 siR-MGP组细胞的凋亡率显著高于D341空白组(P<0.05)。结论MGP在儿童MB癌组织中高表达,下调MGP可以抑制MB细胞的增殖,促进细胞凋亡。
Objective To explore the effect of matrix Gla protein(MGP)on the proliferation,apoptosis of medulloblastoma(MB)cell,and to initially clarify the role and mechanism of MGP in the pathogenesis of MB in children.Methods The MGP gene to be studied in this study was screened through analyzing and comparing the gene expression omnibus database.The cancer tissue and paired paracancerous normal tissue specimens from fourteen MB patients who underwent surgical resection in the Third Affiliated Hospital of Zhengzhou University from January 2018 to December 2021 were collected.The relative expression of MGP protein in cancer tissue and paracancerous normal tissue was detected by reverse transcription-polymerase chain reaction and Western blot.Human MB cells Daoy and D341 were cultured and divided into Doay blank group,Doay small interfering RNA(siR)-MGP group and D341 blank group,D341 siR-MGP group when the cells fusion reached 30%-50%.The cells in Doay siR-MGP group and D341 siR-MGP group were transfected with siR-MGP,while the cells in Doay blank group and D341 blank group were not transfected.At 24,48,72 h after transfection,the proliferation ability of cells in the four groups was detected by cell counting kit-8 method;the apoptosis rate of cells in the four groups was detected at 48 h after transfection by flow cytometry.Results The relative expression of MGP protein in MB cancer tissue was significantly higher than that in the paracancerous normal tissue(P<0.05).At 24,48,72 h after transfection,the proliferation ability of cells in the Doay blank group was significantly higher than that in the Daoy siR-MGP group,and the proliferation ability of cells in the D341 blank group was significantly higher than that in the D341 siR-MGP group(P<0.05).The apoptosis rate of cells in the Doay siR-MGP group was significantly higher than that in the Doay blank group,and the apoptosis rate of cells in the D341 siR-MGP group was significantly higher than that in the D341 blank group(P<0.05).Conclusion MGP is highly expressed in children MB cancer tissue.Down-regulation of MGP can inhibit the proliferation of MB cells and promote cell apoptosis.
作者
李晶晶
宋焕清
冯晓霞
霍志芳
LI Jingjing;SONG Huanqing;FENG Xiaoxia;HUO Zhifang(Department of Paediatrics,the Second Affiliated Hospital of Zhengzhou University,Zhengzhou 450003,Henan Province,China)
出处
《新乡医学院学报》
CAS
2023年第7期608-612,共5页
Journal of Xinxiang Medical University
基金
河南省医学科技攻关联合共建项目(编号:LHGJ20190315)。
关键词
髓母细胞瘤
基质GLA蛋白
增殖
凋亡
medulloblastoma
matrix Gla protein
proliferation
apoptosis
