嗅素结构域家族蛋白3促进MYCN扩增型神经母细胞瘤细胞系体外增殖  

Olfactory domain family protein 3 promotes the proliferation of MYCN-amplified neuroblastoma cell lines in vitro

作　　者：张瑶 张璇 贾安娜 战世佳 郭金鑫 常艳 郭永丽[1] ZHANG Yao;ZHANG Xuan;JIA Anna;ZHAN Shijia;GUO Jinxin;CHANG Yan;GUO Yongli(Beijing Key Laboratory for Pediatric Diseases of Otolaryngology,Head and Neck Surgery,Key Laboratory of Major Diseases in Children,Ministry of Education,Beijing Pediatric Research Institute,Beijing Children′s Hospital,Capital Medical University,National Center for Children′s Health,Beijing 100045,China)

机构地区：[1]国家儿童医学中心,首都医科大学附属北京儿童医院,儿科重大疾病研究教育部重点实验室北京市儿科研究所儿童耳鼻咽喉头颈外科疾病北京市重点实验室,北京100045

出　　处：《基础医学与临床》2025年第2期168-175,共8页Basic and Clinical Medicine

基　　金：北京市自然科学基金(7244341);国家自然科学基金(82141118,82172849)。

摘　　要：目的探究嗅素结构域家族蛋白3(OLFM3)在神经母细胞瘤(NB)中的功能。方法利用R2基因组分析及可视化平台分析OLFM3在NB临床样本中的表达水平和v-myc禽类骨髓细胞瘤病毒癌基因神经母细胞瘤衍生同源基因(MYCN)扩增和患儿预后之间的关系;分析Depmap数据库中OLFM3在不同肿瘤中的表达水平以及OLFM3表达与MYCN扩增的关系;在NB细胞系SK-N-BE(2)、SH-SY5Y和SK-N-AS中利用靶向siRNA干扰OLFM3的表达,RT-qPCR和Western blot检测OLFM3敲低水平;结晶紫染色和实时无标记细胞分析系统观察细胞增殖;克隆形成实验观察细胞克隆形成能力。结果R2数据库分析发现MYCN扩增型NB肿瘤样本的OLFM3表达水平更高(P<0.001);OLFM3高表达组患儿的总生存率明显低于低表达组(P<0.05);Depmap数据库分析发现OLFM3在NB中表达水平较其他肿瘤高,MYCN扩增型细胞系中OLFM3的表达水平高于MYCN非扩增型细胞系(P<0.01);在MYCN扩增型NB细胞中,OLFM3敲低后,细胞增殖能力(P<0.001)和克隆形成能力显著下降(P<0.001),但对MYCN非扩增型细胞无明显影响。结论OLFM3特异性促进MYCN扩增型NB细胞增殖,但对MYCN非扩增型细胞影响较小,是MYCN扩增型高危NB的潜在生物标志物。Objective To explore the function of olfactomedin domain family protein 3(OLFM3)in neuroblastoma(NB).Methods The relationship between the expression of OLFM3 mRNA and v-myc avian myelocytomatosis viral oncogene neuroblastoma derived homolog(MYCN)amplification status and the prognosis of patients in NB clinical samples were clarified by using R2 Genomics Analysis and Visualization Platform.Depmap database was used to examine the expression level of OLFM3 in different tumors cell lines and to identify the correlation between OLFM3 expression and MYCN amplification status in various NB cell lines.RT-qPCR and Western blot were used to detect the knockdown level of OLFM3.Cell proliferation was monitored using crystal violet staining and real-time cellular analysis.The colony formation ability of NB cells was assessed using colony-forming unit assay.Results Analysis of R2 database revealed higher level of OLFM3 expression in MYCN-amplified NB clinical samples(P<0.001).Patients with high OLFM3 expression showed a significantly lower overall survival probability compared to those with low OLFM3 expression(P<0.05).Analysis with Depmap database revealed that the expression level of OLFM3 was higher in NB than that in other kind of tumor.The expression level of OLFM3 was significantly higher in the MYCN-amplified cell lines than in the MYCN-non-amplified cell lines(P<0.01).In MYCN-amplified NB cells,knockdown of OLFM3 inhibited cells proliferation(P<0.001)and colony formation(P<0.001),but there was no noticeable changes observed in MYCN-non-amplified cells.Conclusions OLFM3 specifically promotes the proliferation of MYCN-amplified NB cells,but has a less effect on MYCN-non-amplified cells,indicating it is a potential biomarker for high-risk MYCN-amplified NB.

关 键 词：神经母细胞瘤 v-myc禽类骨髓细胞瘤病毒癌基因神经母细胞瘤衍生同源基因扩增(MYCN) 嗅素结构域家族蛋白3 细胞增殖 

分 类 号：R739.43[医药卫生—肿瘤]