CXC趋化因子受体4和叉头蛋白3基因蛋白及mRNA在儿童纵隔神经母细胞瘤SK-N-SH和LAN-5细胞中的表达及环磷酰胺对其的影响

doi:10.16098/j.issn.0529-1356.2017.05.010

解剖学报 ›› 2017, Vol. 48 ›› Issue (5): 556-560.doi: 10.16098/j.issn.0529-1356.2017.05.010

CXC趋化因子受体4和叉头蛋白3基因蛋白及mRNA在儿童纵隔神经母细胞瘤SK-N-SH和LAN-5细胞中的表达及环磷酰胺对其的影响

谢艳丽王涛*

华中科技大学同济医学院附属武汉儿童医院心胸外科，武汉 430000

收稿日期:2017-03-22 修回日期:2017-04-15 出版日期:2017-10-06 发布日期:2017-10-06
通讯作者: 王涛 E-mail:xieyanli9988@163.com

Expression of CXC chemokine receptor 4 and fork box protein 3 gene protein and mRNA in children mediastinal neuroblastoma cell strains SK-N-SH and LAN-5 and their effects induced by cyclophosphamide

XIE Yan-li WANG Tao*

Cardio-Thoracic Surgery,Wuhan Children’s Hospital（Wuhan Maternal and Child Healthcare)Hospital）, Tongji Medical College, Huazhong University of Science and Technology，Wuhan 430000, China］

Received:2017-03-22 Revised:2017-04-15 Online:2017-10-06 Published:2017-10-06
Contact: WANG Tao E-mail:xieyanli9988@163.com

摘要/Abstract

摘要：

目的观察环磷酰胺对儿童纵隔神经母细胞瘤LAN-5和SK-N-SH细胞CXC趋化因子受体4（CXCR4）和叉头蛋白3（Foxp3）表达的影响。方法采用实时定量聚合酶链反应（Real-time PCR）法检测CXCR4和Foxp3基因mRNA的相对表达，MTT法检测细胞增殖，流式细胞仪分析CXCR4和Foxp3基因的表达。结果 CXCR4和Foxp3在LAN-5和SK-N-SH细胞高表达；环磷酰胺对LAN-5和SK-N-SH细胞的IC50分别为6.5μmol/L和3.9μmol/L；环磷酰胺显著降低了LAN-5细胞表面CXCR4基因蛋白的表达（P＜0.01），也显著降低了SK-N-SH细胞CXCR4 mRNA的表达（P<0.05），同时环磷酰胺显著下调Foxp3基因蛋白（P＜0.05）和Foxp3 mRNA（P<0.01）在LAN-5细胞的表达。结论 CXCR4和Foxp3可能为神经母细胞瘤化疗的潜在靶点。

关键词: 环磷酰胺, 神经母细胞瘤, CXC趋化因子受体4, 叉头蛋白3, 实时定量聚合酶链反应, 儿童

Abstract:

Objective To observe the effect of cyclophosphamide on the CXC chemokin receptor 4(CXCR4) and fork box protein 3(Foxp3) expression of mediastinal neuroblastoma cell strains LAN-5 and SK-N-SH. Methods The Real-time PCR method was used to detect the relative expression of CXCR4 and Foxp3 mRNA, cell proliferation was detected by MTT, CXCR4 and Foxp3 gene expressions were analyzed by flow cytometry. Results CXCR4 and Foxp3 were high expressed in LAN-5 and SK-N-SH cells; IC₅₀ of cyclophosphamide on LAN-5 and SK-N-SH cells were 6.5μmol/L and 3.9μmol/L, respectively; Cyclophosphamide decreased CXCR4 protein expression significantly in LAN-5 cell (P<0.01), also decreased the CXCR4 mRNA expression significantly in SK-N-SH cell (P<0.05), and cyclophosphamide downregulated Foxp3 protein (P<0.05) and Foxp3 mRNA (P<0.01) expression significantly in LAN-5 cells. Conclusion CXCR4 and Foxp3 may be a potential target for neuroblastoma chemotherapy.

Key words: Cyclophosphamide, Neuroblastoma, CXC chemokin receptor 4, Fork box protein 3, Real-time PCR, Child

谢艳丽王涛. CXC趋化因子受体4和叉头蛋白3基因蛋白及mRNA在儿童纵隔神经母细胞瘤SK-N-SH和LAN-5细胞中的表达及环磷酰胺对其的影响[J]. 解剖学报, 2017, 48(5): 556-560.

XIE Yan-li WANG Tao. Expression of CXC chemokine receptor 4 and fork box protein 3 gene protein and mRNA in children mediastinal neuroblastoma cell strains SK-N-SH and LAN-5 and their effects induced by cyclophosphamide[J]. Acta Anatomica Sinica, 2017, 48(5): 556-560.

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CXC趋化因子受体4和叉头蛋白3基因蛋白及mRNA在儿童纵隔神经母细胞瘤SK-N-SH和LAN-5细胞中的表达及环磷酰胺对其的影响

Expression of CXC chemokine receptor 4 and fork box protein 3 gene protein and mRNA in children mediastinal neuroblastoma cell strains SK-N-SH and LAN-5 and their effects induced by cyclophosphamide

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