C12ORF66对MYCN扩增的高危神经母细胞瘤细胞的活性调控

doi:10.16352/j.issn.1001-6325.2024.03.0288

基础医学与临床 ›› 2024, Vol. 44 ›› Issue (3): 288-294.doi: 10.16352/j.issn.1001-6325.2024.03.0288

C12ORF66对MYCN扩增的高危神经母细胞瘤细胞的活性调控

贾安娜^#, 战世佳^#, 张璇, 郭金鑫, 于永波, 郭永丽, 常艳^*

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

收稿日期:2023-11-14 修回日期:2023-12-28 出版日期:2024-03-05 发布日期:2024-02-22
通讯作者: ^*:changyan809@ccmu.edu.cn
作者简介:^#对本文有相同贡献
基金资助:
国家自然科学基金(82141118,82293660/82293665)

Regulatory effect of C12ORF66 on viability of MYCN amplified high-risk neuroblastoma cells

JIA Anna^#, ZHAN Shijia^#, ZHANG Xuan, GUO Jinxin, YU Yongbo, GUO Yongli, CHANG Yan^*

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 Hospital, Capital Medical University, National Center for Children Health, Beijing 100045, China

Received:2023-11-14 Revised:2023-12-28 Online:2024-03-05 Published:2024-02-22
Contact: ^*:changyan809@ccmu.edu.cn

摘要/Abstract

摘要： 目的探究12号染色体开放阅读框66(C12ORF66)对MYCN扩增神经母细胞瘤(NB)细胞系活性的调控效应。方法通过R2数据库GSE16476和GSE49710数据集,分析MYCN扩增和MYCN非扩增NB细胞中C12ORF66表达量,以及C12ORF66表达量与患儿预后的相关性。RT-qRCR检测正常组织永生化细胞系、MYCN扩增及MYCN非扩增细胞系中C12ORF66 mRNA表达差异。构建瞬时敲低和稳定敲低C12ORF66的MYCN扩增细胞株,对照组和敲低组细胞进行实时无标记动态细胞分析(RTCA)、集落形成能力检测及Ki67免疫荧光染色。结果 R2数据库分析显示MYCN扩增NB患儿样本中C12ORF66表达显著高于MYCN非扩增NB患儿样本,并且C12ORF66表达与患儿预后负相关(P＜0.05)。细胞实验表明,与MYCN非扩增细胞系CHLA-255 和SH-SY5Y相比,C12ORF66在MYCN扩增细胞系BE(2)-C和SK-N-BE(2)中表达显著升高(P＜0.001)。瞬时或稳定敲低C12ORF66,MYCN扩增NB细胞增殖显著减缓(P＜0.001),集落形成能力显著降低(P＜0.001),Ki67阳性细胞比例显著减少(P＜0.05)。结论 C12ORF66在MYCN扩增NB患儿样本和细胞系中高表达,且与患儿预后负相关,敲低C12ORF66显著抑制NB细胞活性。

关键词: 12号染色体开放阅读框66, SK-N-BE(2)细胞, MYCN扩增, 细胞增殖

Abstract: Objective To explore the effect of open reading frame 66 (C12ORF66) located at chromosome 12 on the viability of MYCN amplified NB cell lines. Methods DDatasets GSE16476 and GSE49710 in R2 database were analyzed for expression level of C12ORF66 in MYCN amplified and MYCN non-amplified NB cells and its potential correlation with the prognosis of pediatric patients. C12ORF66 mRNA expression level in normal tissue immortalized cell lines, MYCN amplified and MYCN non-amplified cell lines were detected by RT-qRCR. Transient or stable knockdown of C12ORF66 cell lines were constructed to compare the difference in real time cellular analysis (RTCA), colony formation, Ki67 positive cells between the control group and the C12ORF66 knockdown group. Results By analyzing R2 datasets, C12ORF66 level in MYCN amplified samples was significantly higher than that in MYCN non-amplified samples, and the expression of C12ORF66 was negatively correlated with the prognosis of pediatric patients(P＜0.05). C12ORF66 highly expressed in MYCN-amplified BE(2)-C and SK-N-BE(2) cell lines than in MYCN non-amplified CHLA-255 and SH-SY5Y cell lines (P＜0.001). Transient or stable knockdown of C12ORF66 resulted in significant slow down of proliferation of MYCN amplified NB cells (P＜0.001), the colony formation ability was significantly reduced (P＜0.001), and the proportion of Ki67 positive cells was significantly decreased (P＜0.05). Conclusions C12ORF66 was highly expressed in MYCN amplified clinical NB samples and cell lines which is believed to be correlated with poor prognosis of pediatric patients. C12ORF66 knockdown significantly inhibits cell viability of NB cells.

Key words: chromosome 12 open reading frame 66, SK-N-BE(2) cell, MYCN amplified, cell proliferation

中图分类号:

R739.43

贾安娜, 战世佳, 张璇, 郭金鑫, 于永波, 郭永丽, 常艳. C12ORF66对MYCN扩增的高危神经母细胞瘤细胞的活性调控[J]. 基础医学与临床, 2024, 44(3): 288-294.

JIA Anna, ZHAN Shijia, ZHANG Xuan, GUO Jinxin, YU Yongbo, GUO Yongli, CHANG Yan. Regulatory effect of C12ORF66 on viability of MYCN amplified high-risk neuroblastoma cells[J]. Basic & Clinical Medicine, 2024, 44(3): 288-294.

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C12ORF66对MYCN扩增的高危神经母细胞瘤细胞的活性调控

Regulatory effect of C12ORF66 on viability of MYCN amplified high-risk neuroblastoma cells

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