神经生长因子在食管鳞癌细胞中的表达及分泌

doi:10.16098/j.issn.0529-1356.2015.03.010

解剖学报 ›› 2015 ›› Issue (3) : 348-353. DOI: 10.16098/j.issn.0529-1356.2015.03.010

肿瘤生物学

神经生长因子在食管鳞癌细胞中的表达及分泌

李秋霞裴学莲刘洋王玉婷黄燕燕周宗瑶慕晓玲*

作者信息 +

Expression and secretion of nerve growth factor in esophageal squamous cell cancer cell line Eca109

LI Qiu-xia PEI Xue-lian LIU Yang WANG Yu-ting HUANG Yan-yan ZHOU Zong-yao MU Xiao-ling*

Author information +

文章历史 +

摘要

目的探讨神经生长因子（NGF)与食管鳞癌细胞分化的相关性。方法采用有限稀释法分选出圆形和梭形单细胞克隆，采用无血清悬浮培养获得细胞球细胞，贴壁的Eca109细胞分别在含血清和不含血清培养基中培养48h；分别采用反转录-聚合酶链式反应（RT-PCR)技术和免疫印迹法，检测NGF在食管鳞癌细胞中mRNA水平和蛋白水平的表达；免疫荧光技术检测NGF在食管鳞癌细胞中的表达定位；免疫组织化学法检测食管鳞癌组织中NGF的表达定位；用酶联免疫吸附法（ELISA）检测NGF在Eca109细胞培养基中的分泌情况。结果在Eca109细胞中能检测到NGF的mRNA水平和蛋白水平的表达，其中NGF在细胞球细胞中的mRNA水平和蛋白水平的表达量最高。食管癌组织中检测到NGF表达于细胞质。Eca109细胞在无血清培养条件下能够分泌NGF,且明显高于含血清培养基中的含量。结论食管癌细胞系Eca109表达并分泌NGF，并且食管癌组织中表达NGF，NGF可能在维持食管鳞状细胞癌的干细胞特性发挥了重要作用。

Abstract

Objective To investigate the relationship between the nerve growth factor (NGF) and the cell differentiation of esophageal squamous cell cancer. Methods Round and spindle single-cell clones were sorted by limited dilution, and spheres were obtained by cultured in serum-free medium. Adherent Eca109 cells were cultured in serum-containing and serum-free culture medium 48hours, respectively.The expression of NGF mRNA and protein levels in esophageal squamous cell carcinoma (ESCC) were assessed by reverse transcriptase expression of the polymerase chain reaction (RT-PCR) technology and Western blotting analysis, respectively.Immunohistochemical staining determined the nerve growth factor expression in esophageal squamous cell carcinoma specimen. The secretion of NGF in Eca109 was determined by enzyme-linked immunosorbent assay (ELISA). Results The expression of NGF mRNA and protein levels was detected in ESCC,and the spheres cells expressed the highest amount of NGF in mRNA and protein expression level. NGF was expressed in the cytoplasm of ESCC cells in esophageal squamous cell carcinoma specimen. ESCC cells could secreted NGF in serum-free culture conditions, and the NGF secreted was significantly higher than that in serum-containing medium. Conclusion ESCC cells of Eca109 express and secrete NGF, and esophageal squamous cell carcinoma specimen could express nerve growth factor. NGF may play an important role in maintaining ESCC stem cell characteristics.

引用本文

EndNote

Ris (Procite)

Bibtex

导出引用

李秋霞裴学莲刘洋王玉婷黄燕燕周宗瑶慕晓玲*. 神经生长因子在食管鳞癌细胞中的表达及分泌[J]. 解剖学报. 2015(3): 348-353 https://doi.org/10.16098/j.issn.0529-1356.2015.03.010

LI Qiu-xia PEI Xue-lian LIU Yang WANG Yu-ting HUANG Yan-yan ZHOU Zong-yao MU Xiao-ling*. Expression and secretion of nerve growth factor in esophageal squamous cell cancer cell line Eca109[J]. Acta Anatomica Sinica. 2015(3): 348-353 https://doi.org/10.16098/j.issn.0529-1356.2015.03.010

参考文献

［1］Kaplan DR, Miller FD.Neurotrophin signal transduction in the nervous system［J］.Curr Opin in Neurobiol,2000,10(3):381-391.
［2］Chao MV. Neurotrophins and their receptors: a convergence point for many signalling pathways［J］.Nat Rev Neurosci,2003,4(4):299-309.
［3］Sariola H.The neurotrophic factors in non-neuronal tissues［J］. Cell Mol Life Sci,2001,58(8):1061-1066.
［4］Carlo P,Cinzia S,Rossella M,et al.Expression and nerve growth factor receptor on cultured keratinocytes［J］. J Inves Dermatol,1994,103(1):13-18.
［5］Marconi A,Terracina M,Fila C,et al.Expression and function of neurotrophins and their receptors in cultured human keratinocytes［J］. J Invest Dermatol,2003,121(6):1515-1521.
［6］Zhang L,Wang H,Yang Y,et al.NGF induces adult stem Leydig cells to proliferate and differentiate during Leydig cell regeneration［J］. Biochem Biophys Res Commun,2013,436(2):300-305.
［7］Fang D, Nguyen TK, Leishear K, et al. A tumorigenic subpopulation with stem cell properties in melanomas ［ J ］. Cancer Res,2005,65 (20): 9328-9337.
［8］Ponti D, Costa A, Zaffaroni N, et al. Isolati on and in vitro propagation of tumorigenic breast cancer cells with stem /p rogenitor cell properties［ J ］. Cancer Res,2005, 65(13):5506-5511.［9］Cao L,Zhou Y,Zhai B,et al.Sphere-forming cell subpopulations with cancer stem cell properties in human hepatoma cell lines［J］.BMC Gestroenterol,2011,11(1):71.
［10］Zhao ShX,Huang YY,Mu XL.Research into the stem cell characteristics of esophageal carcinoma Eca109 monoclonal cell line［J］.Journal of Shihezi University,2013,31(2):197-201.(in Chinese)
赵生霞,黄燕燕,慕晓玲.食管癌Eca109单克隆细胞株的干细胞特性研究［J］.石河子大学学报,2013,31(2):197-201.
［11］Huang YY,Zhao ShX,Li QX,et al.Isolation and identification of the cancer stem cells from human esophageal squmous cell carcino-ma cell lines Eca109［J］.Journal of Shihezi University,2014,32(3):335-339.(in Chinese)
黄燕燕，赵生霞，李秋霞,等.人食管癌Eca109细胞株中肿瘤干细胞的分离与鉴定［J］.石河子大学学报,2014,32(3):335-339.
［12］Tsunoda S,Okumura T,Ito T,et al.Significance of nerve growth factor overexpression and its autocrine loop in oesophageal squamous cell Carcinoma［J］.Br J Cancer,2006,95(3):322-330.
［13］Truzzi F, Marconi A, Lotti R,et al.Neurotrophins and their receptors stimulate melanoma cell proliferation and migration［J］.J Invest Dermatol,2008,128(8):2031-2040.
［14］Dollé L,Yazidi-Belkoura IE,Adriaenssens E,et al.Nerve growth factor overexpression and autocrine loop in breast cancer cells［J］.Oncogene,2003,22(36):5592-5601.
［15］Molloy NH, Read DE, Gorman AM.Nerve growth factor in cancer cell death and survival［J］.Cancers,2011,3(1):510-530.
［16］Noh SJ, Bae JS, Jamiyandorj U，et al.Expression of nerve growth factor and heme oxygenase-1 predict poor survival of breast carcinoma patients ［J］. BMC Cancer，2013,13:516.
［17］Torrisani J, Buscail L. Molecular pathways of pancreatic carcinogenesis［J］.Ann Pathol,2002,22(5):349-355. 
［18］Zhu ZW,Kleeff J, Kayed H, et al.Nerve growth factor and enhancement of proliferation, invasion, and tumorigenicity of pancreatic cancer cells［J］.Mol Carcinog, 2002,35(3):138-147. 
［19］Okadal Y , Eibl G, Guha S,et al.Nerve growth factor stimulates MMP-2 expression and activity and increases invasio-n by human pancreatic cancer cells［J］.Clinical Expe Metastasis,2004,21(4):285-292.
［20］Tomellini E, Touil Y,lagadecC, et al. NGF and proNGF simultaneously promote symmetric self-renewal, quiescence and EMT to enlarge the breast cancer stem cell compartment［J］. Stem Cells,2014,6(10):1-16.