基因芯片分析孤儿核受体NURR1上调表达对前列腺癌LNCaP细胞基因表达谱的影响

doi:10.3877/cma.j.issn.1674-3253.2022.01.018

目的

探讨孤儿核受体NURR1表达上调对前列腺癌细胞基因表达谱的影响。

方法

构建NURR1过表达载体，通过慢病毒感染方式上调前列腺癌LNCaP细胞中NURR1的表达水平。通过免疫细胞化学染色检测NURR1蛋白的表达，通过基因表达谱芯片分析NURR1过表达组和对照组LNCaP细胞的mRNA、lncRNA表达水平的变化。通过KEEG和GO分析探讨NURR1过表达对相关信号通路和功能的影响。

结果

NURR1基因在LNCaP细胞过表达，与对照组比较共有515个基因发生差异表达，其中上调的mRNA238个，下调的mRNA195个；上调的lncRNA54个，下调的lncRNA28个。差异表达的mRNA和lncRNA主要在于细胞分子功能的改变，其中视网膜结合功能、铁离子结合能力、神经轴突调控以及氧化还原过程、细胞外区域的细胞学组成改变最为显著。细胞因子与细胞因子受体的相互作用以及调节干细胞多能性的有关信号通路都发生显著性改变。

结论

NURR1基因的上调表达对前列腺癌细胞mRNA以及lncRNA表达谱显著改变，可能与其促进前列腺癌进展的功能有关。

关键词: 孤儿核受体, NURR1, 前列腺癌, 基因表达谱

Objective

To investigate the profiling of NURR1-overexpressed LNCaP cells by gene chip analysis.

Methods

NURR1 was ectopic expressed in LNCaP cells by lentivirus infection, validated by immunocytochemical staining. The profiling of mRNA and lncRNA was compared by gene chips between NURR1-overexpressed LNCaP cells and vector-transfected control. KEEG and GO analysis were performed to study the potential functional role and signaling pathways involved.

Results

A total of 515 genes were differentially expressed in NURR1-overexpressed LNCaP cells as compared to its vector-transfected control, including 238 mRNA upregulated, 195 mRNA down regulated, 54 lncRNA upregulated and 28 lncRNA downregulated. The differentially expressed mRNA and lncRNA were mainly involved in molecular functions, among which the retinal binding function, iron ion binding capacity, nerve axon regulation and redox process, and the cytological composition of the extracellular region exhibited the most significance. The interaction between cytokines and cytokine receptors, as well as stem cell pluripotency related signaling pathways were changed significantly.

Conclusion

The mRNA and lncRNA profiling has been significantly changed by the ectopic expression of NURR1, which might play critical role in the development progression of prostate cancer.

Key words: Orphan nuclear receptor, NURR1, Prostate cancer, Gene expression profile

图1 LNCaP细胞NURR1蛋白表达验证以及样品总RNA质检注：图1a为免疫细胞化学检测NUR1在LNCaP细胞中的表达；图1b为凝胶电泳图

图2 差异表达基因分析，差异表达基因火山图

表1 部分显著差异表达基因列表

类别	基因	登记编号	差异倍数	P值
mRNA	ACTG2	NM_001615	40.6	0.0003
	HTR5A	NM_024012	35	0.0007
	MYOD1	NM_002478	34.9	0.0014
	PLOD2	NM_182943	29.7	0
	OR51E1	NM_152430	20.7	0.0004
	CCL20	NM_004591	-11.8	0
	MTRNR2L2	NM_001190470	-11.3	0.0008
	MATN4	NM_003833	-10.6	0.0003
lncRNA	HTR5A-AS1	NR_038945	23	0
	LOC399715	NR_040079	17.7	0.0005
	LOC100129931	NR_033828	12.9	0.0002
	LINC01291	NR_125792	10.4	0.0021
	LOC100996455	NR_073177	10.3	0.0026
	LINC01023	NR_046368	10	0.0016
	LINC00240	NR_026775	-11.1	0.0138
	LOC440896	NR_015361	-9.2	0.0044
	LOC100131047	NR_038401	-8.1	0.0001

表1 部分显著差异表达基因列表

类别	基因	登记编号	差异倍数	P值
mRNA	ACTG2	NM_001615	40.6	0.0003
	HTR5A	NM_024012	35	0.0007
	MYOD1	NM_002478	34.9	0.0014
	PLOD2	NM_182943	29.7	0
	OR51E1	NM_152430	20.7	0.0004
	CCL20	NM_004591	-11.8	0
	MTRNR2L2	NM_001190470	-11.3	0.0008
	MATN4	NM_003833	-10.6	0.0003
lncRNA	HTR5A-AS1	NR_038945	23	0
	LOC399715	NR_040079	17.7	0.0005
	LOC100129931	NR_033828	12.9	0.0002
	LINC01291	NR_125792	10.4	0.0021
	LOC100996455	NR_073177	10.3	0.0026
	LINC01023	NR_046368	10	0.0016
	LINC00240	NR_026775	-11.1	0.0138
	LOC440896	NR_015361	-9.2	0.0044
	LOC100131047	NR_038401	-8.1	0.0001

图3 KEEG分析注：图3a为上调KEEG；图3b为下调KEEG

图4 差异基因GO分析注：图4a为上调表达的基因GO富集分析；图4b为下调表达的基因GO富集分析

图5 差异表达的mRNA和lncRNA的聚类分析注：图5a为差异表达的mRNA；图5b为差异表达的lncRNA，N1~N3分别为NURR1过表达组的3个重复样本，L1~L3为对照组的3个重复样本

[1]	姜涛. 孤儿核受体在前列腺癌中的研究进展[J]. 实用肿瘤学杂志, 2018, 32(4): 377-380.
[2]	Yao H, Yang L, Tian L, et al. LncRNA MSC-AS1 aggravates nasopharyngeal carcinoma progression by targeting miR-524-5p/nuclear receptor subfamily 4 group A member 2 (NR4A2)[J]. Cancer Cell Int, 2020, 20:138.
[3]	Inamoto T, Czerniak BA, Dinney CP, et al. Cytoplasmic mislocalization of the orphan nuclear receptor Nurr1 is a prognostic factor in bladder cancer[J]. Cancer, 2010, 116(2): 340-346.
[4]	Llopis S, Singleton B, Duplessis T, et al. Dichotomous roles for the orphan nuclear receptor NURR1 in breast cancer[J]. BMC Cancer, 2013, 13: 139.
[5]	Karki K, Li X, Jin UH, et al. Nuclear receptor 4A2 (NR4A2) is a druggable target for glioblastomas[J]. J Neurooncol, 2020, 146(1): 25-39.
[6]	Guo J, Zu G, Zhou T, et al. Clinicopathological significance of orphan nuclear receptor Nurr1 expression in gastric cancer[J]. Clin Transl Oncol, 2015, 17(10): 788-794.
[7]	Wang J, Yang J, Zou Y, et al. Orphan nuclear receptor nurr1 as a potential novel marker for progression in human prostate cancer[J]. Asian Pac J Cancer Prev, 2013, 14(3): 2023-2028.
[8]	胡七一, 张建文, 王宏亮, 等. 孤儿核受体NURR1在前列腺癌中的表达以及功能研究[J]. 现代泌尿外科杂志, 2018, 23(4): 299-303.
[9]	张建文, 王宏亮, 邓琼, 等. 核受体RORβ在前列腺癌细胞中的表达及其对肿瘤多细胞球形成的影响[J]. 现代泌尿外科杂志, 2019, 24(8): 667-672.
[10]	王铸, 张建文, 张颖, 等. 草酸钙结石晶体粘附对人肾上皮细胞lncRNA表达谱的影响[J]. 现代泌尿外科杂志, 2018, 23(1): 52-56, 76.
[11]	张建文, 徐长志, 张颖, 等. 草酸钙结石晶体对人近曲肾小管上皮细胞蛋白质表达谱的影响[J/CD]. 中华腔镜泌尿外科杂志（电子版）, 2019, 13(4): 225-229.
[12]	Wu Y, Liu ZG, Shi MQ, et al. Identification of ACTG2 functions as a promoter gene in hepatocellular carcinoma cells migration and tumor metastasis[J]. Biochem Biophys Res Commun, 2017, 491(2): 537-544.
[13]	Edfeldt K, Hellman P, Westin G, et al. A plausible role for actin gamma smooth muscle 2 (ACTG2) in small intestinal neuroendocrine tumorigenesis[J]. BMC Endocr Disord, 2016, 16: 19.
[14]	Schutyser E, Struyf S, Van Damme J. The CC chemokine CCL20 and its receptor CCR6[J]. Cytokine Growth Factor Rev, 2003, 14(5): 409-426.
[15]	Rubie C, Frick VO, Ghadjar P, et al. CCL20/CCR6 expression profile in pancreatic cancer[J]. J Transl Med, 2010, 8: 45.
[16]	Liu B, Jia Y, Ma J, et al. Tumor-associated macrophage-derived CCL20 enhances the growth and metastasis of pancreatic cancer[J]. Acta Biochim Biophys Sin (Shanghai), 2016, 48(12): 1067-1074.

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Profiling study of NURR1-overexpressed LNCaP cells by gene chip analysis

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Profiling study of NURR1-overexpressed LNCaP cells by gene chip analysis