关键词:骨肉瘤;白细胞介素2;逆转录病毒科;基因疗法
【摘要】目的 IL-2基因修饰人骨肉瘤细胞的制备及骨肉瘤患者IL-2、sIL-2R水平测定。方法 用逆转录病毒载体将人IL-2基因插入人骨肉瘤细胞系制备成IL-2基因修饰骨肉瘤细胞;用放射免疫法及ELISA法检测骨肉瘤患者血清内IL-2、sIL-2R水平。结果 经G418筛选和PCR鉴定后制备成功人骨肉瘤IL-2基因修饰细胞,并证实其有分泌IL-2功能,用MTT法测得其培养上清中IL-2表达量为每1×105 细胞24小时50~800u。骨肉瘤患者免疫功能调查发现其血清IL-2水平明显低于正常,而sIL-2R则明显高于正常,术后两者可逐渐正常。结论 制备成功IL-2基因修饰骨肉瘤细胞并发现骨肉瘤患者IL-2水平低下,为进行骨肉瘤基因治疗创造条件。
The Preparation of Human Osteosarcoma Cells Modified by IL- 2 Gene and the Determination of IL- 2 and sIL- 2R in Patients with Osteosarcoma
ZHENG Qiang, FAN Qingyu, GUO Ailin.
Orthpaedic Centre, Tangdu Hospital, Affiliated to The Fourth Military Medical University, Xi’an 710038
【 Abstract 】 Objective To produce human osteosarcoma cells modified by IL- 2 gene and investigate the level of IL- 2 and sIL- 2R in patients with osteosarcoma. Methods The human IL- 2 gene was inserted into human osteosarcoma cells with retroviral vector and the tumor vaccine was produced. IL- 2 and sIL- 2R level in patients with osteosarcoma was determined with radioimmunoassay and ELISA. Results The human osteosarcoma cells modified by IL- 2 gene was produced successfully after G418 resistence selection and analysed by PCR; they could secrete IL- 2 in vitro. The production of IL- 2 was 50~ 800 u per 1× 10 5 cells every 24 h. The data about patients’immune state indicated that IL- 2 was lower than normal control and sIL- 2R higher. Conclusion The success in production of the human osteosarcoma cells modified by Il- 2 gene makes it possisle to carry out further reseach in gene therapy for human osteosarcoma.
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