PLAG1转基因小鼠中差异表达长链非编码RNA的筛选分析

doi:10.19438/j.cjoms.2019.01.004

中国口腔颌面外科杂志 ›› 2019, Vol. 17 ›› Issue (1): 20-25.doi: 10.19438/j.cjoms.2019.01.004

PLAG1转基因小鼠中差异表达长链非编码RNA的筛选分析

徐万林¹, 刘丽敏², 卢浩¹, 杨雯君¹, 沈淑坤¹

1.上海交通大学医学院附属第九人民医院·口腔医学院口腔颌面-头颈肿瘤科,上海 200011;
2.口腔病理科,国家口腔疾病临床医学研究中心,上海市口腔医学重点实验室,上海市口腔医学研究所,上海 200011

收稿日期:2018-07-31 修回日期:2018-09-15 出版日期:2019-01-20 发布日期:2019-02-21
通讯作者: 沈淑坤,E-mail：xwl570127181@163.com
作者简介:徐万林(1989-),男,博士,住院医师,E-mail:450907991@qq.com
基金资助:
国家自然科学基金青年项目（81302359）; 上海市科学技术委员会研究基金项目（16ZR1418800）

Study on the differentially expressed long non-coding RNAs in PLAG1 transgenic mice by gene micro-array

XU Wan-lin¹, LIU Li-min², LU Hao¹, YANG Wen-jun¹, SHEN Shu-kun¹

1.Department of Oromaxillofacial Head and Neck Oncology,Shanghai 200011, China;
2.Department of Oral Pathology, Shanghai Ninth People's Hospital, College of Stomatology, Shanghai Jiao Tong University School of Medicine; National Clinical Research Center for Oral Diseases, Shanghai Key Laboratory of Stomatology & Shanghai Research Institute of Stomatology. Shanghai 200011, China

Received:2018-07-31 Revised:2018-09-15 Online:2019-01-20 Published:2019-02-21

摘要/Abstract

摘要： 目的应用基因芯片技术,筛选PLAG1转基因小鼠下颌下区多形性腺瘤中差异表达的长链非编码RNA（LncRNA）。方法取3对PLAG1转基因小鼠下颌下区多形性腺瘤组织、对侧无瘤腺体组织、空白对照小鼠腺体组织,抽提标本总RNA并反转录合成cDNA探针,与芯片杂交后获得荧光信号, 分析在肿瘤组织中差异表达的长链非编码RNA。随机挑选其中5个LncRNA,利用实时荧光定量PCR在6对样本中验证其表达。采用SPSS13.0软件包中的t检验评价表达差异。结果基因芯片结果显示,与PLAG1转基因小鼠无瘤腺体组织、空白对照小鼠腺体组织相比,PLAG1转基因小鼠肿瘤中共有5627个LncRNA同时发生差异表达,其中2871个上调、2756个下调。实时荧光定量PCR结果显示,5个随机挑选的LncRNA中,4个（AK146794、ENSMUST00000119440、ENSMUST00000140495、Gm12873）与基因芯片结果一致。结论应用基因芯片筛选出PLAG1转基因小鼠中差异表达的长链非编码RNA,为进一步研究多形性腺瘤的发病机制提供了依据。

关键词: 基因芯片, 多形性腺瘤, PLAG1转基因小鼠, 长链非编码RNA

Abstract: PURPOSE: To screen for the differentially expressed long non-coding RNAs (LncRNAs) in pleomorphic adenoma of PLAG1 transgenic mice. METHODS: Three pairs of pleomorphic adenoma, non-tumor gland tissues from PLAG1 transgenic mice and normal gland tissues of submandibular gland from wide-type mice were collected and then the total RNA was extracted. cDNA probe was synthesized by reverse transcription and hybridized as the cDNA microarray for scanning fluorescent signals and differentially expressed LncRNAs. The expressions of 5 randomly selected LncRNAs were validated in another 6 pairs of tissues via real-time PCR. The expression differences were compared and analyzed using SPSS13.0 software package. RESULTS: Microarray results demonstrated that a total of 5627 LncRNAs (2871 upregulated and 2756 downregulated) were differentially expressed in the tumors of PLAG1 transgenic mice. Real-time PCR data showed that the expression of 4 selected LncRNAs (AK146794, ENSMUST00000119440, ENSMUST00000140495 and Gm12873) were consistent with the microarray results. CONCLUSIONS: Analysis of differentially expressed LncRNAs in PLAG1 transgenic mice can provide evidence for pathogenesis of pleomorphic adenoma.

Key words: Microarray, Pleomorphic adenoma, PLAG1 transgenic mice, Long non-coding RNA

中图分类号:

R739.8

徐万林, 刘丽敏, 卢浩, 杨雯君, 沈淑坤. PLAG1转基因小鼠中差异表达长链非编码RNA的筛选分析[J]. 中国口腔颌面外科杂志, 2019, 17(1): 20-25.

XU Wan-lin, LIU Li-min, LU Hao, YANG Wen-jun, SHEN Shu-kun. Study on the differentially expressed long non-coding RNAs in PLAG1 transgenic mice by gene micro-array[J]. China J Oral Maxillofac Surg, 2019, 17(1): 20-25.

参考文献

[1] Tian Z, Li L, Wang L, et al.Salivary gland neoplasms in oral and maxillofacial regions: a 23-year retrospective study of 6982 cases in an eastern Chinese population[J]. Int J Oral Maxillofac Surg, 2010, 39(3): 235-242.
[2] Hu YH, Zhang CY, Xia RH, et al.Prognostic factors of carcinoma ex pleomorphic adenoma of the salivary glands, with emphasis on the widely invasive carcinoma: a clinicopathologic analysis of 361 cases in a Chinese population[J]. Oral Surg Oral Med Oral Pathol Oral Radiol, 2016, 122(5): 598-608.
[3] 夏亮, 汪洋, 胡宇华, 等. 唾液腺恶性多形性腺瘤HER2基因扩增与临床病理特征及预后的关系[J]. 中国口腔颌面外科杂志, 2018, 16(1): 12-19.
[4] Kas K, Voz ML, Roijer E, et al.Promoter swapping between the genes for a novel zinc finger protein and beta-catenin in pleiomorphic adenomas with t(3;8)(p21;q12) translocations[J]. Nat Genet, 1997, 15(2): 170-174.
[5] 赵旭东, 杨雯珺, 王龙, 等. 多形性腺瘤基因1高表达转基因小鼠动物模型的建立[J]. 中华医学遗传学杂志, 2003, 20(5): 390-395.
[6] Shen S, Yang W, Wang Z, et al.Tumor-initiating cells are enriched in CD44(hi) population in murine salivary gland tumor[J]. PLoS One, 2011, 6(8): e23282.
[7] Wang Y, Shang W, Lei X, et al.Opposing functions of PLAG1 in pleomorphic adenoma: a microarray analysis of PLAG1 transgenic mice[J]. Biotechnol Lett, 2013, 35(9): 1377-1385.
[8] Ulitsky I, Bartel DP. lincRNAs: genomics, evolution, and mechanisms[J]. Cell, 2013, 154(1): 26-46.
[9] Zhou JJ, Cheng D, He XY, et al.Knockdown of Hotair suppresses proliferation and cell cycle progression in hepatocellular carcinoma cell by downregulating CCND1 expression[J]. Mol Med Rep, 2017, 16(4): 4980-4986.
[10] Wang ZQ, Cai Q, Hu L, et al.Long noncoding RNA UCA1 induced by SP1 promotes cell proliferation via recruiting EZH2 and activating AKT pathway in gastric cancer[J]. Cell Death Dis, 2017, 8(6): e2839.
[11] Yu T, Shan TD, Li JY, et al.Knockdown of linc-UFC1 suppresses proliferation and induces apoptosis of colorectal cancer[J]. Cell Death Dis, 2016, 7: e2228.
[12] 刘丽敏, 尚文军, 王洋, 等. p53基因敲除PLAG1转基因小鼠模型构建的研究[J]. 临床口腔医学杂志, 2014, 30(3): 149-151.
[13] Xu W, Lu H, Zhu Y, et al.Warthin's tumour in oral and maxillofacial regions: an 18-year retrospective study of 1084 cases in an eastern-Chinese population[J]. Int J Oral Maxillofac Surg, 2018, 47(7): 913-917.
[14] Queimado L, Obeso D, Hatfield MD, et al.Dysregulation of Wnt pathway components in human salivary gland tumors[J]. Arch Otolaryngol Head Neck Surg, 2008, 134(1): 94-101.
[15] Voz ML, Mathys J, Hensen K, et al.Microarray screening for target genes of the proto-oncogene PLAG1[J]. Oncogene, 2004, 23(1): 179-191.
[16] 尚文军, 王洋, 刘丽敏, 等. ERK通路抑制剂苯乙酸钠对转基因小鼠唾液腺肿瘤的作用[J]. 中国口腔颌面外科杂志, 2013, 11(5): 359-364.
[17] Chen Z, Lin S, Li JL, et al.CRTC1-MAML2 fusion-induced lncRNA LINC00473 expression maintains the growth and survival of human mucoepidermoid carcinoma cells[J]. Oncogene, 2018, 37(14): 1885-1895.

PLAG1转基因小鼠中差异表达长链非编码RNA的筛选分析

Study on the differentially expressed long non-coding RNAs in PLAG1 transgenic mice by gene micro-array

PDF (PC)

可视化

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 14

编辑推荐

Metrics

本文评价

[1]	许飞虎, 徐万林, 刘胜文, 丁修明, 张祥, 杨雯君. 腮腺区复发性多形性腺瘤恶变侵犯颧骨1例报告[J]. 中国口腔颌面外科杂志, 2021, 19(6): 573-576.
[2]	何优雅, 张誉, 戴振霖, 林承重, 曹巍, 季彤. 长链非编码RNA CASC15在成釉细胞瘤中的差异表达及生物信息学分析[J]. 中国口腔颌面外科杂志, 2021, 19(5): 411-416.
[3]	孙琦, 马超, 董敏俊, 陶晓峰. 感兴趣区大小选择对多形性腺瘤和腺淋巴瘤ADC测量的影响[J]. 中国口腔颌面外科杂志, 2021, 19(1): 29-35.
[4]	苑绪光, 朱骏飞, 李天竹, 光梦凯, 张晔. 同侧腮腺同时存在2种不同肿瘤1例报告[J]. 中国口腔颌面外科杂志, 2021, 19(1): 94-96.
[5]	周光明, 梁衍灿, 黄子贤, 张彬. 长链非编码RNA FOXD2-AS1对舌鳞癌细胞迁移、侵袭的影响[J]. 中国口腔颌面外科杂志, 2020, 18(6): 495-500.
[6]	汪洋, 夏亮, 胡宇华, 张春叶, 王丽珍, 李江, 田臻. 基于RNA-Seq分析挖掘唾液腺多形性腺瘤核心基因及通路[J]. 中国口腔颌面外科杂志, 2019, 17(5): 397-402.
[7]	徐万林, 朱云, 卢浩, 刘丽敏, 沈淑坤, 刘胜文, 杨雯君. 15例下颌下腺复发性多形性腺瘤临床病理分析[J]. 中国口腔颌面外科杂志, 2019, 17(5): 465-468.
[8]	魏军水, 卢浩, 杨雯君, 沈淑坤, 徐万林. 人多形性腺瘤中与cyclin D1共表达LncRNA的初步筛选及鉴定[J]. 中国口腔颌面外科杂志, 2019, 17(3): 198-203.
[9]	夏亮, 汪洋, 胡宇华, 王丽珍, 顾挺, 李江, 田臻. HER2基因在唾液腺恶性多形性腺瘤细胞增殖、周期、凋亡中的作用[J]. 中国口腔颌面外科杂志, 2018, 16(4): 310-316.
[10]	夏亮, 胡宇华, 王丽珍, 顾挺, 李江, 田臻. 唾液腺恶性多形性腺瘤中CDH1基因启动子甲基化与E-cadherin表达的相关性分析[J]. 中国口腔颌面外科杂志, 2018, 16(2): 120-125.
[11]	夏亮, 汪洋, 胡宇华, 顾挺, 李江, 石慧敏, 田臻. 唾液腺恶性多形性腺瘤HER2基因扩增与临床病理特征及预后的关系[J]. 中国口腔颌面外科杂志, 2018, 16(1): 12-19.
[12]	邓刚，李晓光，何悦，于德栋. 复发性腮腺多形性腺瘤的手术治疗及并发症预防[J]. 中国口腔颌面外科杂志, 2017, 15(3): 227-229.
[13]	李孔亮, 张珊珊, 杨宏宇, 方征宇. MEG3长链非编码RNA在舌鳞癌组织中的表达及意义[J]. 中国口腔颌面外科杂志, 2016, 14(6): 514-517.
[14]	夏亮, 张春叶, 胡宇华, 钱佳骏, 李江, 田臻. 启动子甲基化状况对唾液腺恶性多形性腺瘤体外细胞系细胞E-cadherin表达的影响[J]. 中国口腔颌面外科杂志, 2016, 14(4): 308-314.