蚯蚓纤溶酶增加人食管癌细胞ECA109对放射线敏感性的实验研究-东南大学学报医学版

蚯蚓纤溶酶增加人食管癌细胞ECA109对放射线敏感性的实验研究

单位：1.东南大学附属中大医院,消化科,江苏,南京,210009； 2.东南大学医学院附属南京第二医院,肿瘤放疗科,江苏,南京,210003； 3.南京农业大学,免疫生化研究所,江苏,南京,210095； 4.东南大学附属中大医院,血液科,江苏,南京,210009

关键词：蚯蚓纤溶酶放射增敏食管癌细胞株ECA109 硒谷胱甘肽过氧化物酶

分类号：R815, R735.1

出版年·卷·期（页码）：2007·26·第六期（409-413）

摘要：

目的:探讨蚯蚓纤溶酶(EFE)对食管癌细胞的放射增敏作用及机制.方法:选取人食管癌细胞株ECA109,采用MTT法观察EFE对细胞的生长抑制作用,克隆形成实验观察EFE对细胞的放射增敏作用,流式细胞术观察EFE对细胞周期分布的影响,RT-PCR法检测EFE对细胞内硒谷胱甘肽过氧化物酶(SeGPx)mRNA表达的影响.结果:EFE对ECA109有生长抑制作用,其作用呈剂量依赖性,IC50为3.8 ukU·ml-1;EFE 0.1、0.3 ukU·ml-1的放射增敏比分别为1.37和1.59;EFE作用24 h后,食管癌细胞G2与M期比例明显下降,细胞内SeGPx基因表达明显下调.结论:EFE在体外可抑制ECA109的生长,同时可增加其对放射线的敏感性;放射增敏机制可能与EFE清除细胞G2/M期阻滞及下调细胞内抗氧化酶SeGPx mRNA表达有关.

Objective To study the radiosensitivity-enchancing effect of earthworm fibrinolytic enzyme(EFE) on human esophageal carcinoma cell line and its related mechanism.Methods The esophageal carcinoma cell line EC A109 was used in the experiment.MTT assay was performed to analyze the inhibitory ratio of the cell line,the clonogenic assay was used to determine cell survival fraction,the flow cytometer was used to check cell cycle distribution and the RT-PCR was used to determine the gene expression of selenium dependent glutathione peroxidase(SeGPx).Results The proliferation of cell line ECA 109 was suppressed by EFE in vitro.The inhibitory concentration of 50%(IC50) was 3.8 ukU·ml-1.EFE could definitely enhance radiosensitivity of ECA 109 cells.The sensitivity-enchancement-ratio was 1.37 for 0.1 ukU·ml-1EFE and 1.59 for 0.3 ukU·ml-1 of EFE respectively.EFE abrogated the G2/M arrest induced by γ-ray.Meanwhile,the expression of SeGPx mRNA decreased after the EFE or(and) γ-ray treatment.Conclusion EFE might be a potential radiosensitizer for esophageal carcinoma.In addition,the abrogation of G2/M arrest and the downregulated expression of SeGPx mRNA may be related with EFE’s radiosensitizing effect.

参考文献：

[1] 张绍章, 田琼, 王克为. 中药地龙胶囊对食管癌和肺癌的放射增效作用, 1992(3)
[2] 赵锐, 纪建国, 童朋元. 赤子爱胜蚓中抗肿瘤与纤溶酶原激活活性蛋白质的分离与鉴定. 生物化学与生物物理学报(英文版)2002(5)
[3] OLIVARES-FONTT E, BESCHIN A, van DIJCK E. Trypanosoma cruzi is lysed by coelomic cytolytic factor-1 an invertebrace analogue of rumor necrosis factor,and induces phenoloxidase activity in coelomic fluid of Eisenia foetida, 2002(1)
[4] 朱锋荣, 何执中. 蚯蚓活性成分的抗肿瘤作用. 药物生物技术2006(4). doi:10.3969/j.issn.1005-8915.2005.04.018
[5] 陈洪. "地龙2号"抗肿瘤作用的初步观察. 临床肿瘤学杂志2001(4). doi:10.3969/j.issn.1009-0460.2001.04.024
[6] 何道伟, 陈洪. 蚯蚓提取物体内和体外抗肿瘤作用的研究. 中国生化药物杂志2005(6). doi:10.3969/j.issn.1005-1678.2005.06.011
[7] CHEN H, SHOICHI T, MICHIO I. Earthworm fibrinolytic enzyme:anti-tumor activity on human hepatoma cells in vitro and in vivo. 2007(10)
[8] 余艳秋, 陈洪, 黄锦. 蚯蚓纤溶酶抑制胃癌细胞株MGC803增殖及诱导凋亡的作用. 东南大学学报(医学版)2007(1). doi:10.3969/j.issn.1671-6264.2007.01.004
[9] 袁玲, 徐建民, 周元聪. 纯化的蚯蚓蛋白对多种血液系统瘤细胞系的作用. 中国临床医学2004(2). doi:10.3969/j.issn.1008-6358.2004.02.019
[10] 孙新臣, 王俊杰, 甄永苏. 粉防己碱对放射线的增敏作用与机理研究. 中华放射医学与防护杂志2004(2). doi:10.3760/cma.j.issn.0254-5098.2004.02.003
[11] REE A H, BRATLAND A, NOME R V. Inhibitory targeting of checkpoint kinase signaling overrides radiation-induced cell cycle gene regulation:a therapeutic strategy in tumor cell radiosensitization?. 2004(3). doi:10.1016/j.radonc.2004.07.002
[12] 惠周光, 李晔雄, 杨伟志. UCN-01增加鼻咽癌细胞放射敏感性的相关机制. 中华放射肿瘤学杂志2006(1)
[13] ABOU-BEDAIR F A, HORI H, NOGASAWA H. Comparison of hypoxic cell radiosensitizers,KIN-804,KIN-844,KIN-806 and TX-1877,on brain and liver metabolizing capacities in mice bearing ehrlich ascites carcinoma. 2002(5)
[14] JORDAN B F, SONVEAUX P, FERON O. Nitric oxide as a radiosensitizer:evidence for an intrinsic role in addition to its effect on oxygen delivery and consumption. 2004(5). doi:10.1002/ijc.20046
[15] MINCHINTON A I, ROJAS A, SMITH K A. Glutathione depletion in tissues after administriation of buthionine sulphoximine, 1984(8)
[16] 耿长新, 曾昭冲. 多西他赛对人肝癌细胞的放疗增敏作用及其作用机制探讨. 复旦学报(医学版)2003(3). doi:10.3969/j.issn.1672-8467.2003.03.020
[17] 唐小云, 梁再赋, 贾秀坤. 地龙提取物对小鼠腹腔MФ和脾细胞NO及TNF-α产生的影响. 细胞与分子免疫学杂志2003(5)
[18] GRDISA M, POPOVIC M, HRZENJAK T. Glycolipoprotein extract(G-90) from earthworm Eisenia foetida exerts some antioxidative activity, 2001(4)

服务与反馈：

【文章下载】【发表评论】【查看评论】【加入收藏】

提示：您还未登录，请登录！点此登录