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.

参考文献 12

[1]	Qiu B, Matthay KK. Advancing therapy for neuroblastoma[J]. Nat Rev Clin Oncol, 2022, 19: 515-533.
[2]	Yu EY, Cheung NV, Lue NF, et al. Connecting telomere maintenance and regulation to the developmental origin and differentiation states of neuroblastoma tumor cells[J]. J Hematol Oncol, 2022, 15:117-133.
[3]	谭正兰,何晓燕,陈卉,等. miR-221促进神经母细胞瘤SH-SY5Y细胞N-MYC表达和细胞增殖[J]. 基础医学与临床,2016,36:1329-1334.
[4]	Cai Y, Xu G, Wu F, et al. Genomic alterations in PIK3CA-mutated breast cancer result in mTORC1 activation and limit the sensitivity to PI3Kα inhibitors[J]. Cancer Res, 2021, 81: 2470-2480.
[5]	Tang X, Zhang Y, Wang G, et al. Molecular mechanism of adenosylmethionine sensing by SAMTOR in mTORC1 signaling[J]. Sci Adv, 2022, 8: 3868. doi: 10.1126/sciadv.abn3868.
[6]	Liu GY, Sabatini DM. mTOR at the nexus of nutrition, growth, ageing and disease[J]. Nature reviews. Nat Rev Mol Cell Biol, 2020, 21: 183-203.
[7]	Abdul Aziz NA, Mokhtar NM, Harun R, et al. A 19-Gene expression signature as a predictor of survival in colorectal cancer[J]. BMC Med Genomics, 2016, 9. doi: 10.1186/s12920-016-0218-1.
[8]	Zeineldin M, Patel AG, Dyer MA, et al. Neuroblastoma: When differentiation goes awry[J]. Neuron, 2022, 110: 2916-2928.
[9]	Matthay KK, Maris JM, Schleiermacher G, et al. Neuroblastoma[J]. Nat Rev Dis Primers, 2016, 2. doi: 10.1038/nrdp.2016.78.
[10]	Durbin AD, Zimmerman MW, Dharia NV, et al. Selec-tive gene dependencies in MYCN-amplified neuroblastoma include the core transcriptional regulatory circuitry[J]. Nat Genet, 2018, 50: 1240-1246.
[11]	Thompson D, Vo KT, London WB, et al. Identification of patient subgroups with markedly disparate rates of MYCN amplification in neuroblastoma: a report from the International Neuroblastoma Risk Group project[J]. Cancer, 2016, 122: 935-945.
[12]	Wolfson RL, Chantranupong L, Wyant GA, et al. KICSTOR recruits GATOR1 to the lysosome and is necessary for nutrients to regulate mTORC1[J]. Nature, 2017, 543: 438-442.

[1]	梁香存, 魏小雨, 梁健, 王庆, 耿广. 安罗替尼通过调控miR-16-5p/PD-1轴抑制人非小细胞肺癌细胞系增殖并促进凋亡[J]. 基础医学与临床, 2024, 44(4): 503-512.
[2]	陆凯, 陆建菊, 郭文利, 黄建棋, 李志华. lncRNA VIM-AS5在人乳腺癌细胞系中的表达及对乳腺癌细胞增殖和迁移的影响[J]. 基础医学与临床, 2024, 44(4): 447-453.
[3]	李娜, 李涛, 杨冬梨, 姚媛. 和厚朴酚抑制人脂肪间充质干细胞增殖[J]. 基础医学与临床, 2024, 44(4): 483-488.
[4]	廖镇城, 杨思懿, 吴平安. 提高m⁶A去甲基化酶FTO表达抑制鼻咽癌细胞增殖[J]. 基础医学与临床, 2024, 44(1): 57-62.
[5]	潘闪, 林铿强, 陈树兴. 培美曲塞抑制人肺腺癌细胞系A549增殖[J]. 基础医学与临床, 2023, 43(9): 1359-1363.
[6]	梁辉勇, 韦秋慧, 丁亚, 彭小青, 姚静, 苏齐鉴. 虫草素抑制TNF-α诱导的人角质形成细胞系HaCaT增殖[J]. 基础医学与临床, 2023, 43(9): 1390-1393.
[7]	张培波, 李义. 沙棘总黄酮抑制膀胱癌细胞系T24增殖[J]. 基础医学与临床, 2023, 43(7): 1122-1126.
[8]	侯孟杰, 赵娜, 黄富强, 王亚南, 刘芳铭. 载脂蛋白E通过降低高尔基体膜蛋白73表达抑制小鼠肝癌细胞系增殖[J]. 基础医学与临床, 2023, 43(5): 762-770.
[9]	王芳芳, 曹慧媛, 汪婧, 王宁, 黄国良. 上调miR-200c-5p的表达抑制人结直肠癌细胞系SW480增殖[J]. 基础医学与临床, 2023, 43(2): 241-245.
[10]	刘爱霞, 张浩, 孙杰, 杨丽华, 赵晓涛. GPS2对肝癌细胞系MHCC-97H增殖、迁移与侵袭的调控作用[J]. 基础医学与临床, 2023, 43(12): 1784-1791.
[11]	张珍珍, 李洋, 高生宝, 夏德鑫, 严玉兰. 重组新城疫病毒rL-RVG抑制人肺腺癌细胞系增殖[J]. 基础医学与临床, 2023, 43(10): 1549-1556.
[12]	沈永华, 汪峻峰, 程泽星, 冯娟娟. 秦皮甲素抑制人鼻咽癌细胞系HNE-3增殖[J]. 基础医学与临床, 2022, 42(8): 1237-1242.
[13]	江宁, 郭钧, 刘铖, 周举. 莪术醇抑制人骨肉瘤细胞系的增殖和促凋亡[J]. 基础医学与临床, 2022, 42(8): 1200-1205.
[14]	陈建苗. DFG-STYK1/NOK对小鼠胚胎成纤维细胞系NIH3T3增殖以及细胞周期G₁/S的影响[J]. 基础医学与临床, 2021, 41(6): 859-864.
[15]	刘亚琪, 郑承红. miR-205通过靶向调控Wnt-5a抑制乳头状甲状腺癌的细胞增殖[J]. 基础医学与临床, 2021, 41(3): 358-363.

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

PDF

可视化

摘要/Abstract

引用本文

使用本文

参考文献 12

相关文章 15

编辑推荐

Metrics

本文评价