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目的建立小鼠肿瘤模型,并用体内成像的方法进行检测,降低传统方法中的系统误差与人为测量错误,提高模型的准确性与客观性。方法构建萤火虫素酶的慢病毒表达载体,体外包装能表达萤火虫素酶的慢病毒颗粒。用病毒颗粒转染目标细胞,筛选出稳定表达萤火虫素酶的目标细胞系。体外运用双荧光素酶分析系统进行质控检测,通过后建立小鼠肿瘤模型,最后使用活体成像的方法对小鼠肿瘤模型中肿瘤的大小进行定量分析,以评估小鼠肿瘤模型的恶化程度。结果成功构建含有萤火虫素酶基因的慢病毒质粒;包装能正确表达具有功能活性萤火虫素酶的慢病毒颗粒;使用病毒颗粒感染目标细胞并筛选出能表达萤火虫素酶的稳转细胞系;建立小鼠皮下肿瘤模型,并利用活体成像的方法对肿瘤大小进行定量分析。结论通过该种方法可以建立表达萤火虫素酶的稳转细胞系,利用体内成像的检测方法可以准确的对肿瘤模型的发生发展进行评估。
OBJECTIVE: To establish a tumor model in mice and to detect it by in vivo imaging methods to reduce the systematic errors and human measurement errors in traditional methods and to improve the accuracy and objectivity of the model. Methods The lentivirus vector of firefly enzyme was constructed, and the lentivirus particles expressing firefly enzyme were packaged in vitro. The target cells are transfected with the virus particles and the target cell line stably expressing the fireflyrase is screened out. The dual luciferase assay system was used for quality control testing in vitro. After establishment of the mouse tumor model, the tumor size in the mouse tumor model was quantitatively analyzed by live imaging to evaluate the degree of tumor deterioration in the mouse model. Results The lentiviral plasmid containing fireflyrase gene was constructed successfully. The recombinant lentiviral plasmid was successfully packaged and expressed correctly. The virus particles were used to infect the target cells and the stable cell lines expressing fireflyrase were screened out. Mouse subcutaneous tumor model, and the use of live imaging methods for quantitative analysis of tumor size. Conclusion This method can be used to establish a stable cell line that expresses firefly enzyme and the in vivo imaging method can accurately evaluate the occurrence and development of tumor model.