[白细胞介素18转染的骨髓基质细胞对大鼠颅内胶质瘤生长的影响]
[Effect of bone marrow stromal cells transfected with interleukin 18 on growth of intracranial glioma in rats].
作者信息
Li Chun-Hui, Jiao Bao-Hua
机构信息
Department of Neurosurgery, The Second Affiliated Hospital, Hebei Medical University, Shijiazhuang, Hebei, 050000, P. R. China.
出版信息
Ai Zheng. 2007 Jan;26(1):38-43.
BACKGROUND & OBJECTIVE: Because of the invasion and immune escape characteristics, surgical resection, radiotherapy, and chemotherapy had no definite curative effects on intracranial glioma. Because bone marrow stromal cells (BMSCs) can track migrating cells, and interleukin 18 (IL-18) can enhance antitumor immune reaction, this study was to explore the effect of IL-18-transfected BMSCs on growth of glioma in rats.
METHODS
Pure BMSCs were obtained by culturing rat bone marrow cells and identified by flow cytometry (FCM). BMSCs were transfected with retrovirus LXSN/IL-18 to prepare BMSCs/IL-18. IL-18 genetic transcription and expression were assessed by reverse transcription-polymerase chain reaction (RT-PCR) and immunofluorescence. The characteristic variations of BMSCs were detected by MTT assay, FCM, and immunofluorescence. The effect of BMSCs/IL-18 on activation of T cells was evaluated by ELISA. Glioma-bearing rats were divided into BMSCs group, BMSCs/IL-18 group, PBS group, and control group and received relevant treatments. The effect of BMSCs/IL-18 on growth of glioma was observed.
RESULTS
IL-18 gene was expressed stably in BMSCs/IL-18. The proliferation speed of BMSCs/IL-18 was slower than that of BMSCs. The secretion of interferon-gamma (IFN-gamma) from rat spleen lymphocytes in BMSCs/IL-18 group was 9 times more than that in BMSCs group. Tumor volume was (18.26+/-6.84) mm(3) in BMSCs group, (6.37+/-1.52) mm(3) in BMSCs/IL-18 group, (22.48+/-6.02) mm(3) in PBS group, and (21.06+/-5.83) mm(3) in control group; survival time of the rats was (25.3+/-6.4) days, (84.7+/-16.3) days, (21.6+/-4.7) days, and (22.5+/-6.2) days, respectively. After transplantation of BMSCs/IL-18, CD4(+) T cells in glioma were increased to 37.7+/-3.5 and CD8(+) T cells were increased to 32.3+/-4.5 in each field of view (x200).
CONCLUSION
BMSCs/IL-18 could express IL-18 gene stably, and have definite therapeutic effect on glioma in rats.
背景与目的
由于颅内胶质瘤具有侵袭和免疫逃逸特性，手术切除、放疗及化疗对其疗效均不确切。鉴于骨髓基质细胞（BMSCs）具有追踪迁移细胞的能力，且白细胞介素18（IL-18）能增强抗肿瘤免疫反应，本研究旨在探讨经IL-18转染的BMSCs对大鼠胶质瘤生长的影响。
方法
通过培养大鼠骨髓细胞获取纯化的BMSCs，并采用流式细胞术（FCM）进行鉴定。用逆转录病毒LXSN/IL-18转染BMSCs制备BMSCs/IL-18。采用逆转录聚合酶链反应（RT-PCR）和免疫荧光法评估IL-18基因的转录和表达。通过MTT法、FCM和免疫荧光检测BMSCs的特性变化。采用ELISA法评估BMSCs/IL-18对T细胞活化的影响。将荷瘤大鼠分为BMSCs组、BMSCs/IL-18组、PBS组和对照组并给予相应处理，观察BMSCs/IL-18对胶质瘤生长的影响。
结果
IL-18基因在BMSCs/IL-18中稳定表达。BMSCs/IL-18的增殖速度比BMSCs慢。BMSCs/IL-18组大鼠脾淋巴细胞分泌的干扰素-γ（IFN-γ）比BMSCs组多9倍。BMSCs组肿瘤体积为（18.26±6.84）mm³，BMSCs/IL-18组为（6.37±1.52）mm³，PBS组为（22.48±6.02）mm³，对照组为（21.06±5.83）mm³；大鼠生存时间分别为（25.3±6.4）天、（84.7±16.3）天、（21.6±4.7）天和（22.5±6.2）天。移植BMSCs/IL-18后，胶质瘤中CD4⁺T细胞在每个视野（×200）增加至37.7±3.5，CD8⁺T细胞增加至32.3±4.5。
结论
BMSCs/IL-18可稳定表达IL-18基因，对大鼠胶质瘤有确切治疗作用。
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