基于活体成像的表达人PSCA抗原的小鼠肿瘤模型的建立

In vivo Bioluminescence Imaging of Luc Labeled Human PSCA-expressed Tumor in Mouse Model

目的:探讨并建立可供药物评价或生物学功能研究的表达人PSCA抗原的荧光素酶小鼠肿瘤模型。方法:克隆人PSCA基因及荧光素酶基因luc,构建真核表达质粒pcDNA-PSCA及pcDNA-luc,共转染RM-1细胞株;照度计检测转染细胞的荧光素酶活性,RT-PCR及流式细胞术检测PSCA的表达,筛选出共表达荧光素酶及人PSCA的RM-PSCA/luc细胞株;将RM-PSCA/luc细胞接种C57BL/6小鼠,观察所致肿瘤的生长情况及小鼠存活状况,并对小鼠进行活体成像检测Luc的表达;采用免疫组织化学染色方法检测人PSCA在小鼠肿瘤组织中的表达。结果:筛选到了稳定共表达Luc及人PSCA抗原的RM-PSCA/luc细胞,接种实验小鼠全部成瘤,肿瘤生长迅速,小鼠平均存活38天;转染荧光素酶基因的肿瘤细胞生物特性稳定,活体成像仪检测到转染荧光素酶细胞在小鼠体内活体成像,荧光素酶表达活性的强弱能够反映肿瘤大小;免疫组织化学染色方法检测到小鼠肿瘤组织PSCA的表达。结论:成功构建了可用于活体成像表达人PSCA抗原的小鼠肿瘤模型,为相关肿瘤药物及疫苗的研发奠定基础。

Objectives ：To establish a tumor model in mouse with expressing Luc and human PSCA for the development of new anti-tumor drugs or vaccines. Methods：The Luc and human PSCA genes were cloned and pcDNA-luc and pcDNA-PSCA plasmids were constructed. RM-1 cells were stably transfected with both pcDNA- luc and pcDNA-PSCA plasmids. The Luc-expressed cells were examined with luminometer and the PSCA- expressed cells were examined with RT-PCR and flow cytometry analysis. The male C57BL/6 mice were inoculated with RM-PSCA/Luc cells, then the growth and the survival time of mice were measured, respectively. In vivo bioluminescenee imaging was used to detect Luc expression and immunohistochemistry analysis was used to detect PSCA expression. Results：RM-PSCA/Lue cells stably expressing Luc and PSCA were obtained and can establish tumor growth in 100% of inoculated mice. Conclusion ：The Lue-expressed and PSCA-expressed tumor model in mice was successfully established and could be used for evaluating the activities of drugs or vaccines.