【摘要】：目的利用慢病毒感染的方法建立能穩(wěn)定表達(dá)前列腺特異性膜抗原(PSMA)和熒光素酶的PSMA~+-PC3細(xì)胞系,并將其接種到裸鼠皮下檢測其成瘤能力。方法包裝表達(dá)PSMA的慢病毒和表達(dá)熒光素酶的慢病毒,將其感染PC3細(xì)胞后,用嘌呤霉素進(jìn)行篩選,建立能夠穩(wěn)定表達(dá)PSMA和熒光素酶的PSMA~+ -PC3細(xì)胞和僅表達(dá)熒光素酶的PSMA--PC3細(xì)胞;利用流式細(xì)胞術(shù)檢測PSMA的表達(dá)情況。將PSMA~+ -PC3和PSMA~- -PC3細(xì)胞接種于裸鼠皮下建立荷瘤模型,利用生物發(fā)光成像法觀察其在體內(nèi)的成瘤情況,利用免疫組織化學(xué)染色法檢測PSMA~+ -PC3和PSMA~- -PC3腫瘤組織中PSMA的表達(dá)情況。結(jié)果成功建立了能夠穩(wěn)定表達(dá)PSMA和熒光素酶的PSMA~+ -PC3細(xì)胞和僅表達(dá)熒光素酶的PSMA~- -PC3細(xì)胞,流式細(xì)胞術(shù)檢測證實(shí)PSMA~+ -PC3細(xì)胞上PSMA呈陽性表達(dá),小動物活體成像結(jié)果顯示PSMA~+ -PC3和PSMA~- -PC3細(xì)胞均可有效成瘤,免疫組織化學(xué)染色檢測證實(shí)PSMA~+ -PC3腫瘤上PSMA的表達(dá)為陽性。結(jié)論成功建立了能穩(wěn)定表達(dá)PSMA和熒光素酶的PSMA~+ -PC3細(xì)胞系,并在裸鼠體內(nèi)證實(shí)其可有效成瘤,為靶向PSMA的研究提供了有用的細(xì)胞和動物模型。
[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.
【作者單位】： 第四軍醫(yī)大學(xué)免疫學(xué)教研室;第四軍醫(yī)大學(xué)西京醫(yī)院泌尿外科;
【基金】：國家重點(diǎn)基礎(chǔ)研究發(fā)展計(jì)劃(973)(2013CB530500) 國家自然科學(xué)基金(81372225)
【分類號】：R737.25
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本文編號：2224395