Walker William A, Tarannum Mubin, Vivero-Escoto Juan L
Department of Chemistry, University of North Carolina at Charlotte, Charlotte NC 28223, U.S.A.; The Center for Biomedical Engineering and Science, University of North Carolina at Charlotte, Charlotte NC 28223, U.S.A.
The Center for Biomedical Engineering and Science, University of North Carolina at Charlotte, Charlotte NC 28223, U.S.A.; Nanoscale Science Program, University of North Carolina at Charlotte, Charlotte NC 28223, U.S.A.
J Mater Chem B. 2016 Feb 21;4(7):1254-1262. doi: 10.1039/C5TB02079D. Epub 2016 Jan 7.
In this study, mesoporous silica nanoparticles (MSNs) were functionalized with Cholera toxin subunit B (CTxB) protein to influence their intracellular trafficking pathways. The CTxB-MSN carrier was synthesized, and its chemical and structural properties were characterized. Endocytic pathway inhibition assays showed that the uptake of CTxB-MSNs in human cervical cancer (HeLa) cells was partially facilitated by both chlatrin- and caveolae-mediated endocytosis mechanisms. Laser scanning confocal microscopy (LSCM) experiments demonstrated that CTxB-MSNs were taken up by the cells and partially trafficked through the trans-Golgi network into to the endoplasmic reticulum in a retrograde fashion. The delivery abilities of CTxB-MSNs were evaluated using propidium iodide, an impermeable cell membrane dye. LSCM images depicted the release of propidium iodide in the endoplasmic reticulum and cell nucleus of HeLa cells.
在本研究中,用霍乱毒素B亚基(CTxB)蛋白对介孔二氧化硅纳米颗粒(MSNs)进行功能化修饰,以影响其细胞内运输途径。合成了CTxB-MSN载体,并对其化学和结构性质进行了表征。内吞途径抑制试验表明,网格蛋白介导和小窝介导的内吞机制均部分促进了人宫颈癌(HeLa)细胞对CTxB-MSNs的摄取。激光扫描共聚焦显微镜(LSCM)实验表明,CTxB-MSNs被细胞摄取,并部分通过反式高尔基体网络逆行运输到内质网中。使用碘化丙啶(一种不能透过细胞膜的染料)评估了CTxB-MSNs的递送能力。LSCM图像描绘了HeLa细胞内质网和细胞核中碘化丙啶的释放情况。