Safety markers for rhabdomyosarcoma cells using an in vivo imaging system
Safety markers for rhabdomyosarcoma cells using an in vivo imaging system
复制标题
使用体内成像系统进行横纹肌肉瘤细胞的安全标记。
DOI：
10.3892/ol.2018.8789
复制
发表时间：
2018-07-01
期刊：
影响因子：
2.9
通讯作者：
Li, Feng
中科院分区：
文献类型：
--
作者：
Du, Shutong;Meng, Lian;Li, Feng
体内成像系统是一种新兴的、发展迅速的技术，在生命科学领域有着广泛的应用，包括细胞示踪。IVIS能够通过计算从标本发出的光子数量来量化生物事件，包括肿瘤增殖。将PLA 802增强的绿色荧光蛋白（EGFP）、PLA 802单体樱桃荧光蛋白（mCherry）、RH 30-EGFP和RH 30-mCherry肿瘤细胞皮下注射到18只BALB/c雌性裸小鼠中，每100 μl注射5×106个细胞，以定量分析IVIS追踪的EGFP和mCherry细胞。倒置荧光显微镜显示PLA 802-EGFP（95.3±1.2%）和PLA 802-mCherry（95.8±1.7%）之间以及RH 30-EGFP（94.7±2.1%）和RH 30-mCherry（95.2±1.9%）之间的转染效率没有差异。转染对PLA 802和RH 30的细胞形态没有影响。慢病毒-EGFP和慢病毒-mCherry转染前后的细胞迁移、侵袭和增殖试验结果显示无显著差异。因此，慢病毒-EGFP和慢病毒-mCherry可作为PLA 802和RH 30细胞的安全生物学标记。体内实验证明，慢病毒-EGFP和慢病毒-mCherry肿瘤发光信号在所有小鼠中通过IVIS观察到。苏木精-伊红染色和免疫组化结果显示，PLA 802-EGFP、PLA 802-mCherry、RH 30-EGFP和RH 30-mCherry细胞系具有与母细胞相似的横纹肌肉瘤（RMS）特征。综上所述，mCherry和绿色荧光蛋白在人RMS PLA 802和RH 30癌细胞中可以在体外和体内长期安全稳定地表达。
In vivo imaging system (IVIS) is a novel and rapidly expanding technology that is widely applied in life sciences, including cell tracing. IVIS is able to quantify biological events, including tumor proliferation, through counting the number of photons emitted from a specimen. PLA802-enhanced green fluorescent protein (EGFP), PLA802-monomeric cherry fluorescent protein (mCherry), RH30-EGFP and RH30-mCherry tumor cells were injected into 18 BALB/c female nude mice subcutaneously with 5×106 cells in 100 µl to quantitatively analyze EGFP and mCherry cells traced by IVIS. Inversion fluorescence microscopy revealed no transfection efficiency difference between PLA802-EGFP (95.3±1.2%) and PLA802-mCherry (95.8±1.7%), or between RH30-EGFP (94.7±2.1%) and RH30-mCherry (95.2±1.9%). Transfection did not influence the cell morphology of PLA802 or RH30. The cell migration, invasion and proliferation assay results of lentivirus-EGFP and lentivirus-mCherry revealed no significant difference prior to or following transfection. Therefore, lentivirus-EGFP and lentivirus-mCherry may serve as safety biological markers for PLA802 and RH30 cells. In vivo experiments demonstrated that lentivirus-EGFP and lentivirus-mCherry tumor luminescence signals were observed in all mice by IVIS. Hematoxylin-eosin staining and immunohistochemistry indicated that PLA802-EGFP, PLA802-mCherry, RH30-EGFP and RH30-mCherry cell lines exhibited rhabdomyosarcoma (RMS) characteristics like the maternal cells. In summary, mCherry and green fluorescent protein in human RMS PLA802 and RH30 cancer cells may be safely and stably expressed for a long time in vitro and in vivo.