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*

doi:10.16098/j.issn.0529-1356.2017.05.010

PDF(380 KB)

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

肿瘤生物学

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*

作者信息 +

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*

Author information +

文章历史 +

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。

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.

导出引用

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 https://doi.org/10.16098/j.issn.0529-1356.2017.05.010

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 https://doi.org/10.16098/j.issn.0529-1356.2017.05.010

参考文献

［1］Zhang DW, Jin M, Zeng Q, et al. Posterior mediastinal neuroblastoma in 35 children ［J］. Journal of Applied Clinical Pediatrics, 27(11): 835-836. (in Chinese)

张大伟，金眉，曾骐，等. 后纵隔神经母细胞瘤35例［J］.实用儿科临床杂志，2012，27（11）：835-836.

［2］Zhang Y, Zhang WL, Huang DSh, et al. Clinical features and outcomes of neuroblastoma patients aged above 5 years ［J］. Chinese Journal of Contemporary Pediatrics, 2016,18(12): 1217-1221. (in Chinese)

张谊，张伟令，黄东生，等. 5岁以上神经母细胞瘤患者的临床特点及疗效分析［J］.中国当代儿科杂志，2016，18（12）：1217-1221.

［3］Sivapalan N, Wattie J, Inman MD, et al. Lung-homing of endothe-lial progenitor cells and airway vascularization is only partially de-pendant on eosinophils in a house dust mite-exposed mouse model of allergic asthma ［J］. PLoS One, 2014, 9(10): e109991.

［4］Niu KT, Mo BW, Wang ZhX, et al. Research progress of CXCL12-CXCR4/CXCR7 signal axis related diseases ［J］. Shandong Medical Journal, 2015, 55(17): 89-91. (in Chinese)

牛坤汀，莫碧文，王志霞，等. CXCL12-CXCR4/CXCR7信号轴相关疾病研究进展［J］.山东医药，2015，55（17）：89-91.

［5］Yang Sh, Tang YY. Pivotal effects of FOXP3 in modulating immune response ［J］. Immunological Journal, 2016, 32(3): 262-268. (in Chinese)

杨爽，汤玉瑜. FOXP3在调节免疫应答中的作用［J］.免疫学杂志，2016，32（3）：262-268.

［6］Mc Murchy AN, Gillies J, Gizzi MC, et al. A novel function for FOXP3 in humans: intrinsic regulation of conventional T cells［J］. Blood, 2013, 121(8): 1265-1275.

［7］Han M, Lv S, Zhang Y. The prognosis and clinicopathology of CXCR4 in gastric cancer patients: a meta-analysis ［J］. Tumour Biol, 2014, 35(5): 4589-4597.

［8］Srivastava S, Koch LK, Campbell DJ. IFNalphaR signaling in effector but not regulatory Tcells is required for immune dysregulation during type I IFN-dependent inflammatory disease ［J］. J Immunol, 2014, 193(6):2733-2742.

［9］Ryu NG, Moon IJ, Chang YS, et al. Cochlear implantation for profound hearing loss after multimodal treatment for neuroblastoma in Children ［J］. Clin Exp Otorhinolaryngol, 2015, 8(4): 329-334.

［10］Karlsson J, Ora I, PornAres I, et al. Arsenic trioxide-induced death of neuroblastoma cells involves activation of bax and does not require p53 ［J］. Clin Cancer Res, 2004, 10(9): 3179-3188.

［11］Qi K, Li Y, Xiong XL, et al. Arsenic trioxide potentiates the cytotoxicity of chemotherapeutic agents against human SK-N-SH neuroblastoma cell line［J］. Journal of China Pediatric Blood and Cancer, 2016, 21(5): 252-256. (in Chinese)

亓凯，黎阳，熊稀霖，等. 三氧化二砷联合化疗药物对神经母细胞瘤SK-N-SH细胞系杀伤作用的研究［J］. 中国小儿血液与肿瘤杂志，2016，21（5）：252-256.

［12］Sharp SE, Gelfand MJ, Shulkin BL. Pediatrics: diagnosis ofneuroblastoma ［J］. Semin Nucl Med, 2011, 41(5): 345-353.

［13］Peled A, Tavor S. Role of CXCR4 in the pathogenesis of acute myeloid leukemia ［J］. Theranostics, 2013, 3(1): 34-39.

［14］Tian XH, Bai ShL, Fan J, et al. Down-regulation of tankyrase 1 mediated by RNAi leads to the apoptosis of SH-SY5Y cells ［J］. Acta Anatomica Sinica, 2014, 45(2): 196-203. (in Chinese)

田晓红，柏树令，范军，等.RNA干扰介导的端锚聚合酶 1 表达下调诱导人神经母细胞瘤细胞的凋亡［J］.解剖学报，2014，45（2）：196-203.

［15］Huang CQ, Chen JL, Zhu DD, et al. Advances in the study of Foxp3 transcriptional regulation［J］. Journal of Pathogen Biology, 2016, 11(5): 3-6. (in Chinese)

黄彩裙，陈金铃，朱丹丹，等. Foxp3 转录调控的研究进展［J］. 中国病原生物学杂志，2016，11（5）：3-6.

［16］Patton DT, Wilson MD, Rowan WC, et al. The PI3K p110delta regulates expression of CD38 on regulatory T cells ［J］. PLoS One, 2011, 6(3): e17359.