Shahbazi Sepideh, Haghighipour Nooshin, Soleymani Sepehr, Nadji Seyed Alireza, Bolhassani Azam
Department of Hepatitis and AIDs, Pasteur Institute of Iran, Tehran, Iran.
National Cell Bank of Iran, Pasteur Institute of Iran, Tehran, Iran.
Biotechnol Lett. 2018 Jun;40(6):923-931. doi: 10.1007/s10529-018-2551-2. Epub 2018 Apr 9.
In this study, transfection efficiency of human papillomavirus (HPV) E7 DNA and protein constructs into HEK-293T normal cell line, and A549 and TC-1 tumor cell lines was evaluated by four delivery systems including supercharge GFP, hPP10 cell penetrating peptide, TurboFect and Lipofectamine using fluorescence microscopy and flow cytometry.
The results indicated that Lipofectamine 2000 and TurboFect produced more effective transfection for GFP and E7-GFP DNA constructs in HEK-293T cells compared to in A549 and TC-1 cells (p < 0.05). In contrast, the supercharge GFP was efficient for E7 DNA and E7 protein delivery in both normal cell (~ 83.94 and ~ 77.01% for HEK-293T), and cancer cells (~ 71.69 and ~ 67.19% for TC-1, and ~ 73.86 and ~ 67.49% for A549), respectively. Indeed, in these cell lines, transfection efficiency by +36 GFP reached ~ 60-80%. Moreover, the hPP10 produced the best transfection result for E7-GFP protein in HEK-293T cells (~ 63.66%) compared to TurboFect (~ 32.95%); however, the efficiency level of hPP10 was only ~ 17.51 and ~ 16.36% in TC-1 and A549 cells.
Our data suggested that the supercharge GFP is the most suitable transfection vehicle for DNA and protein delivery into TC-1 and A549 tumor cell lines compared to other carriers.
在本研究中,使用荧光显微镜和流式细胞术,通过包括超电荷绿色荧光蛋白(supercharge GFP)、人多聚脯氨酸10(hPP10)细胞穿透肽、TurboFect和脂质体转染试剂(Lipofectamine)在内的四种递送系统,评估人乳头瘤病毒(HPV)E7 DNA和蛋白构建体转染至人胚肾293T(HEK-293T)正常细胞系以及A549和TC-1肿瘤细胞系的效率。
结果表明,与A549和TC-1细胞相比,脂质体转染试剂2000(Lipofectamine 2000)和TurboFect对HEK-293T细胞中的绿色荧光蛋白(GFP)和E7-GFP DNA构建体产生了更有效的转染(p < 0.05)。相比之下,超电荷绿色荧光蛋白在正常细胞(HEK-293T中分别约为83.94%和77.01%)和癌细胞(TC-1中分别约为71.69%和67.19%,A549中分别约为73.86%和67.49%)中对E7 DNA和E7蛋白递送均有效。实际上,在这些细胞系中,+36 GFP的转染效率达到约60 - 80%。此外,与TurboFect(约32.95%)相比,hPP10对HEK-293T细胞中的E7-GFP蛋白产生了最佳转染结果(约63.66%);然而,hPP10在TC-1和A549细胞中的效率水平仅约为17.51%和16.36%。
我们的数据表明,与其他载体相比,超电荷绿色荧光蛋白是将DNA和蛋白递送至TC-1和A549肿瘤细胞系的最合适转染载体。