不同转移潜能MG63骨肉瘤细胞亚株裸鼠肺转移模型的建立

Establishment of a Nude Mouse Model of MG63 Osteosarcoma Cell Sublines with Different Potentials of Metastasis

目的建立不同转移潜能的骨肉瘤MG63细胞亚株裸鼠肺转移模型,为探讨骨肉瘤发生发展机制奠定实验基础。方法MTT法检测不同转移潜能的人骨肉瘤MG63细胞亚株M8、M6细胞增殖情况,将生长良好的细胞制成2×107/ml细胞悬液注射裸鼠尾静脉,第4周起处死裸鼠,肉眼及镜下观察肺部转移灶形成情况,肺组织经过石蜡包埋切片后行苏木精-伊红染色观察病理变化,并采用免疫组化方法检测肿瘤相关指标Cyclin E1和Bcl-2的表达。结果高转移潜能细胞M8组细胞增殖能力强于M6组。尾静脉注射瘤细胞后,M8组20只裸鼠除2只死亡外,有17只发生肺转移;低转移潜能M6组20只裸鼠仅1只发生肺转移。免疫组化结果显示M8组裸鼠肺组织Cyclin E1和Bcl-2表达呈阳性,而M6组未见阳性细胞表达。结论采用尾静脉注射人骨肉瘤细胞亚株M8、M6可成功建立高、低转移潜能的骨肉瘤裸鼠肺转移模型,为深入研究骨肉瘤侵袭和转移机制及寻求有效治疗方法提供了实验平台。

Objective To establish a nude mouse lung metastasis model of human osteosarcoma MG63 cell sublines with different potentials of metastasis in order to explore the mechanism of osteosarcoma development. Methods The proliferation of human osteosarcoma cell sublines M8 and M6 was determined by MTT assay. 2×10^7 cells of M8 and M6 sublines were injected into 20 nude mice via tail vein respectively. Mice were sacrificed starting on the 4th week after injection, and lung metastases were evaluated under both macroscopic and microscopic observations with HE staining. The expression of Bcl-2 and Cyclin E1 in lung metastases was detected by immunohistochemistry. Results The growth of high-metastatic sublines M8 was much faster than low-metastatic subline M6. Lung metastases occurred in 17 mice injected with M8, while only 1 mouse injected with M6. Moreover, the expression of Bcl-2 and cyclin E1 in lung metastatic tissues were positive. Conclusion A nude mouse lung metastasis model with high-or low-metastatic potentials can be established by injecting human osteosarcoma cell sublines M8 and M6 via tail vein respectively, which provides an experimental foundation for further studying the mechanism of osteosarcoma invasion and metastasis.