摘要
目的：探讨骨髓间充质干细胞（Mesenchymal Stem cells,MSCs）的全细胞抗原（WCAs）对于人肺腺癌细胞株A549移植瘤的预防接种作用,为发掘肿瘤免疫治疗提供新策略。方法：采用全骨髓贴壁法原代培养小鼠MSCs并以流式细胞仪鉴定,取3~5代MSCs细胞以15Gy X线灭活获取（Whole cell antigens,WCAs）,以该抗原皮下接种于BALB/c小鼠（1次/3 d,共2周）,获得免疫接种小鼠模型,对照组皮下注射同体积的PBS;为了方便示踪,以Lipofectamine TM 2000细胞脂质体转染绿色荧光蛋白（GFP）,获得标记GFP-A549细胞株,皮下移植瘤株进行荷瘤,采用小动物荧光成像仪观察肿瘤生长;同时评估肿瘤直径并计算肿瘤体积,行Ki-67免疫组化染色初步分析肿瘤增殖能力。结果：肺腺癌GFP-A549细胞株皮下移植成功使小鼠荷瘤,实验组小鼠的肿瘤体积显著小于对照组（★P〈0.05,★★P〈0.01）,小动物荧光成像仪结果与解剖结果一致;实验组Ki-67的表达低于对照组。结论：本研究采用MSCs获得WCAs在荷瘤前对小鼠进行免疫刺激,发现其确实有抑制肿瘤生长的作用。
Objective：To estimate the whole cell antigens （WCAs） of Mesenchymal Stem cells （MSCs） could generate immune response against cancer in mice which was induced by lung cancer cell line A549,in hope of finding new method in cancer immunotherapy.Methods：The MSCs was isolated and cultured adherent cells from marrow,and then was to irradiate to X-ray （15Gy） to get the WCAs （1/3 days,2 weeks）.The mice were challenged with A549 cells.The control mice immunized subcutaneously with PBS.Stably transfect A549 cells with a green fluorescent protein （GFP） by LipofectamineTM 2000 to enable to be traced.The growth of tumor was monitored by small animal vivo imaging system.Both the volume and diameter of those tumors was measured.The expression of Ki-67 was monitored by immunohistochemisty.Results：Tumors grew progressively in the PBS immunized group （★P〈0.05,★★P〈0.01） both in small animal vivo imaging system and volume assessment.Strikingly,the expression Ki-67 was down-regulated in experiments groups.Conclusion：We found that vaccination of mice with MSCs generated consistent anticancer effect of lung carcinoma.
出处
《现代生物医学进展》
CAS
2015年第4期610-613,621,共5页
Progress in Modern Biomedicine
基金
国家自然科学基金青年科学基金项目(81102678)
