低剂量环磷酰胺对Lewis肺癌小鼠CD4+CD25+Treg细胞功能的影响

doi:国家自然科学基金资助课题（30571724）

低剂量环磷酰胺对Lewis肺癌小鼠CD4⁺CD25⁺Treg细胞功能的影响

李欣¹,崔永生^1,2, 刘白楠¹, 李一¹

1.吉林大学基础医学院免疫学教研室，吉林长春 130021；2.吉林大学第一医院胸外科，吉林长春 130021

收稿日期:2007-10-09 修回日期:1900-01-01 出版日期:2008-09-28 发布日期:2008-09-28
通讯作者: 李一

Effect of low dose cyclophosphamide on function of CD4⁺CD25⁺Treg cells of mice with Lewis lung cancer

LI Xin¹,CUI Yong-sheng^1,2,LIU Bai-nan¹,LI Yi¹

1.Department of Immunology,School of Basic Medical Sciences,Jilin University,Changchun 130021,China; 2.Department of Thoracic Surgery,First Hospital,Jilin University,Changchun 130021,China

Received:2007-10-09 Revised:1900-01-01 Online:2008-09-28 Published:2008-09-28
Contact: LI Yi

摘要/Abstract

摘要： 目的：研究低剂量环磷酰胺（CTX）单次注射对Lewis肺癌小鼠CD4⁺CD25⁺Treg细胞功能的影响，探讨CD4⁺CD25⁺Treg 细胞与肿瘤发生、发展的关系。方法：将传代培养的Lewis肺癌细胞接种于C57BL/6小鼠右腋皮下，建立Lewis肺癌模型，随机分成3组：CTX组、肿瘤组、单纯对照组。采用CTX单次注射，观测各组肿瘤体积动态变化；采用流式细胞术检测各组小鼠脾脏CD4⁺CD25⁺Treg细胞数量变化；采用半定量RT-PCR方法检测各组小鼠脾脏Foxp3 mRNA表达水平；采用MTT法检测脾脏T淋巴细胞增殖功能；采用LDH释放法检测脾脏特异性细胞毒性T淋巴细胞(CTLs)细胞的杀伤活性。结果：与未治疗肿瘤组相比,CTX治疗可延缓荷19 d Lewis肺癌小鼠的肿瘤生长；CTX组小鼠脾脏CD4⁺CD25⁺Treg细胞数量低于肿瘤组（P< 0.05）；CTX组小鼠脾脏Foxp3 mRNA表达水平明显低于肿瘤组（P< 0.05）；CTX组小鼠脾脏T淋巴细胞功能明显高于肿瘤组（P<0. 05）；CTX组小鼠脾脏T淋巴细胞杀伤活性比肿瘤组略高，但变化不显著（P>0.05）。结论：CTX单次注射可降低Lewis肺癌小鼠脾脏CD4⁺CD25⁺Treg细胞的数量及Foxp3 mRNA表达，使T淋巴细胞增殖功能及脾细胞中CTLs杀伤功能增强。

关键词: CD4^+<, sup>CD25^+<, sup>调节性T细胞, 监测, 免疫学

Abstract: Abstract:Objective To study the effect of low dose cyclophosphamide on function of CD4⁺CD25⁺Treg of mice with Lewis lung cancer and investigate the relationship between CD4⁺CD25⁺Treg and the genesis and development of tumor.Methods The models were divided into three groups:CTX group,tumor group and control group.In CTX group, models were established by injection CTX (25 mg•kg^-1) and subcultivated Lewis lung cancer cells were injected subcutaneously to the right axilla of C57BL/6 mice 7 d later.The dynamic changes of tumor volume were observed.The changes of the number of CD4⁺CD25⁺Treg were detected by flow cytometer and the expression of Foxp3 mRNA in spleen was analyzed by semi-quantitative RT-PCR.The changes of T lymphocyte proliferation in spleen were detected by MTT test.The changes of T lymphocyte killing function in spleen were detected by LDH releasing test.Results Compared with tumor group,CTX treatment might delay the development of tumor in 19 d-mice with Lewis lung cancer.The number of CD4⁺CD25⁺ Treg in spleen of mice in CTX group was lower than that in tumor group (P<0.05).The expression of Foxp3 mRNA in spleen lymphocyte in CTX group was significantly lower than that in tumor group (P<0.05).The changes of T lymphocyte proliferation in spleen in CTX group were significantly higher than that in tumor group (P<0.05).The changes of T lymphocyte killing function in spleen in tumor group was not significantly lower than that in CTX group (P>0.05). Conclusion After CTX injection,the number of CD4⁺CD25⁺Treg and the expression of Foxp3 mRNA in spleen of mice with Lewis lung cancer decrease,the immunological function of mice with Lewis lung cancer increase.It can provid experiment evidence for the tumor immunotherapy aimed directly at CD4⁺CD25⁺Treg.

Key words: CD4^+<, sup>CD25^+<, sup> regulatory T cell, monitorying, immunologic

中图分类号:

R734.2

李欣,崔永生, 刘白楠, 李一. 低剂量环磷酰胺对Lewis肺癌小鼠CD4⁺CD25⁺Treg细胞功能的影响[J]. J4, 2008, 34(5): 733-736.

LI Xin,CUI Yong-sheng,LIU Bai-nan,LI Yi. Effect of low dose cyclophosphamide on function of CD4⁺CD25⁺Treg cells of mice with Lewis lung cancer[J]. J4, 2008, 34(5): 733-736.

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