过表达前列腺特异性膜抗原和荧光素酶的PC3稳定细胞系的建立及成瘤性鉴定
发布时间：2018-09-05 13:20
【摘要】：目的利用慢病毒感染的方法建立能稳定表达前列腺特异性膜抗原(PSMA)和荧光素酶的PSMA~+-PC3细胞系,并将其接种到裸鼠皮下检测其成瘤能力。方法包装表达PSMA的慢病毒和表达荧光素酶的慢病毒,将其感染PC3细胞后,用嘌呤霉素进行筛选,建立能够稳定表达PSMA和荧光素酶的PSMA~+ -PC3细胞和仅表达荧光素酶的PSMA--PC3细胞;利用流式细胞术检测PSMA的表达情况。将PSMA~+ -PC3和PSMA~- -PC3细胞接种于裸鼠皮下建立荷瘤模型,利用生物发光成像法观察其在体内的成瘤情况,利用免疫组织化学染色法检测PSMA~+ -PC3和PSMA~- -PC3肿瘤组织中PSMA的表达情况。结果成功建立了能够稳定表达PSMA和荧光素酶的PSMA~+ -PC3细胞和仅表达荧光素酶的PSMA~- -PC3细胞,流式细胞术检测证实PSMA~+ -PC3细胞上PSMA呈阳性表达,小动物活体成像结果显示PSMA~+ -PC3和PSMA~- -PC3细胞均可有效成瘤,免疫组织化学染色检测证实PSMA~+ -PC3肿瘤上PSMA的表达为阳性。结论成功建立了能稳定表达PSMA和荧光素酶的PSMA~+ -PC3细胞系,并在裸鼠体内证实其可有效成瘤,为靶向PSMA的研究提供了有用的细胞和动物模型。
[Abstract]:Objective to establish a PSMA~ PC3 cell line which can stably express prostate specific membrane antigen (PSMA) and luciferase by lentivirus infection, and then inoculate it into nude mice for subcutaneous detection of its tumorigenesis ability. Methods Lentivirus expressing PSMA and lentivirus expressing luciferase were packaged. After infected with PC3 cells, the cells were screened by purine mycin to establish PSMA~ PC3 cells and PSMA--PC3 cells expressing luciferase only and stable expression of PSMA and luciferase. The expression of PSMA was detected by flow cytometry. PSMA~ PC3 and PSMA~- PC3 cells were inoculated into nude mice subcutaneously to establish tumor-bearing model. The tumor formation in vivo was observed by bioluminescent imaging, and the expression of PSMA in PSMA~ PC3 and PSMA~- PC3 tumor tissues was detected by immunohistochemical staining. Results PSMA~ PC3 cells expressing PSMA and luciferase stably and PSMA~- PC3 cells expressing luciferase only were successfully established. The positive expression of PSMA on PSMA~ PC3 cells was confirmed by flow cytometry. In vivo imaging of small animals showed that PSMA~-PC3 and PSMA~- PC3 cells were all effective in tumorigenesis. The expression of PSMA on PSMA~-PC3 tumor was confirmed by immunohistochemical staining. Conclusion PSMA~-PC3 cell line which can stably express PSMA and luciferase was successfully established and proved to be effective in tumorigenesis in nude mice. It provides a useful cell and animal model for the study of targeted PSMA.
【作者单位】： 第四军医大学免疫学教研室;第四军医大学西京医院泌尿外科;
【基金】：国家重点基础研究发展计划(973)(2013CB530500) 国家自然科学基金(81372225)
【分类号】：R737.25
，
本文编号：2224395
[Abstract]:Objective to establish a PSMA~ PC3 cell line which can stably express prostate specific membrane antigen (PSMA) and luciferase by lentivirus infection, and then inoculate it into nude mice for subcutaneous detection of its tumorigenesis ability. Methods Lentivirus expressing PSMA and lentivirus expressing luciferase were packaged. After infected with PC3 cells, the cells were screened by purine mycin to establish PSMA~ PC3 cells and PSMA--PC3 cells expressing luciferase only and stable expression of PSMA and luciferase. The expression of PSMA was detected by flow cytometry. PSMA~ PC3 and PSMA~- PC3 cells were inoculated into nude mice subcutaneously to establish tumor-bearing model. The tumor formation in vivo was observed by bioluminescent imaging, and the expression of PSMA in PSMA~ PC3 and PSMA~- PC3 tumor tissues was detected by immunohistochemical staining. Results PSMA~ PC3 cells expressing PSMA and luciferase stably and PSMA~- PC3 cells expressing luciferase only were successfully established. The positive expression of PSMA on PSMA~ PC3 cells was confirmed by flow cytometry. In vivo imaging of small animals showed that PSMA~-PC3 and PSMA~- PC3 cells were all effective in tumorigenesis. The expression of PSMA on PSMA~-PC3 tumor was confirmed by immunohistochemical staining. Conclusion PSMA~-PC3 cell line which can stably express PSMA and luciferase was successfully established and proved to be effective in tumorigenesis in nude mice. It provides a useful cell and animal model for the study of targeted PSMA.
【作者单位】： 第四军医大学免疫学教研室;第四军医大学西京医院泌尿外科;
【基金】：国家重点基础研究发展计划(973)(2013CB530500) 国家自然科学基金(81372225)
【分类号】：R737.25
，
本文编号：2224395
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