Internalization Characterization of Si Nanorod with Camouflaged Cell Membrane Proteins Reveals ATXN2 as a Negative Regulator
Abstract
:1. Introduction
2. Materials and Methods
2.1. Fabrication of Nanorods
2.2. Characterization of SiNRs by SEM and TEM
2.3. Cell Culture
2.4. Cell Membrane Protein Extraction
2.5. SDS-PAGE and Silver Staining
2.6. Western Blot
2.7. LC-MS/MS
2.8. GO Enrichment Analysis
2.9. Membrane Protein Coating and Labeling
2.10. Quantitative Analysis on Nanorod Uptake
2.11. Immunostaining, EGF Binding Assay, Colocalization Assay, and Confocal Microscopy
2.12. Total RNA Isolation and qRT-PCR Analysis
2.13. Small Interfere RNA Transfection and Chemical Inhibitors Studies
2.14. Statistic Method
3. Results
3.1. Fabrication of Size-Controllable SiNR Array
3.2. Nanofabrication of SiNR Coating with Membrane Protein
3.3. Proteomics Analysis of Coating Membrane Proteins Derived from Five Cancer Cell Lines
3.4. SiNR Internalization Efficiency Is also Determined by Target Cells
3.5. Gene Ontology on Cancer Cell Internalization
3.6. ATXN2 Is a Negative Regulator of Nanorod Internalization
4. Discussion
Supplementary Materials
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
References
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SiNR | Diameter (μm) | Length (μm) |
---|---|---|
L-size | 0.412 ± 0.027 | 3.495 ± 0.181 |
M-size | 0.409 ± 0.010 | 2.229 ± 0.049 |
S-size | 0.105 ± 0.011 | 0.656 ± 0.126 |
Cell line | Genes |
---|---|
Hela | GNAI1 CAV2 SYAP1 LYN JAK1 |
MDA231 | SAA1 LOXL2 VTN AHI1 HIP1 THBS1 TGFB2 ITGA5 FAM129B EIF3E |
PDX | MELTF SRC |
MCF7 | BCL2 GSK3B BAX RHOT2 |
Panc-1 | ATXN2 VASP ARPC5L ARPC5 DCTN1 PXN LIMA1 MYH10 |
Cell Type | GO ID | GO Term | Adjust p-Value |
---|---|---|---|
Panc-1 | 0001725 | stress fiber | 6.87 × 10−4 |
0003779 | actin binding | 1.06 × 10−4 | |
0005916 | fascia adherens | 7.08 × 10−4 | |
0007015 | actin filament organization | 7.46 × 10−4 | |
0008064 | regulation of actin polymerization or depolymerization | 1.36 × 10−3 | |
0008154 | actin polymerization or depolymerization | 1.78 × 10−3 | |
0030041 | actin filament polymerization | 4.13 × 10−3 | |
0030833 | regulation of actin filament polymerization | 2.83 × 10−3 | |
0030838 | positive regulation of actin filament polymerization | 2.86 × 10−3 | |
0045806 | negative regulation of endocytosis | 5.65 × 10−3 | |
0051495 | positive regulation of cytoskeleton organization | 2.49 × 10−3 | |
0097517 | contractile actin filament bundle | 6.87 × 10−4 | |
1902903 | regulation of supramolecular fiber organization | 4.18 × 10−3 | |
1902905 | positive regulation of supramolecular fiber organization | 4.21 × 10−4 | |
Panc-PDX | 0031579 | membrane raft organization | 5.72 × 10−3 |
0034446 | substrate adhesion-dependent cell spreading | 1.90 × 10−3 | |
0035635 | entry of bacterium into host cell | 7.88 × 10−3 | |
1900024 | regulation of substrate adhesion-dependent cell spreading | 8.61 × 10−4 | |
MCF-7 | 1905710 | positive regulation of membrane permeability | 5.32 × 10−5 |
MDA231 | 0019898 | extrinsic component of membrane | 6.14 × 10−3 |
0051015 | actin filament binding | 4.10 × 10−3 | |
0005178 | integrin binding | 3.51 × 10−3 | |
0005884 | actin filament | 1.13 × 10−3 | |
0006898 | receptor-mediated endocytosis | 1.93 × 10−3 | |
0050840 | extracellular matrix binding | 8.92 × 10−3 | |
0033627 | cell adhesion mediated by integrin | 7.46 × 10−3 | |
0048259 | regulation of receptor-mediated endocytosis | 2.20 × 10−3 | |
0048260 | positive regulation of receptor-mediated endocytosis | 1.96 × 10−3 | |
Hela | 0031234 | extrinsic component of cytoplasmic side of plasma membrane | 4.50 × 10−3 |
0044406 | adhesion of symbiont to host | 5.66 × 10−3 |
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Lu, Y.; Dai, J.; Kong, N.; Liu, J.; Gong, J.; Yao, Y. Internalization Characterization of Si Nanorod with Camouflaged Cell Membrane Proteins Reveals ATXN2 as a Negative Regulator. Cells 2019, 8, 931. https://doi.org/10.3390/cells8080931
Lu Y, Dai J, Kong N, Liu J, Gong J, Yao Y. Internalization Characterization of Si Nanorod with Camouflaged Cell Membrane Proteins Reveals ATXN2 as a Negative Regulator. Cells. 2019; 8(8):931. https://doi.org/10.3390/cells8080931
Chicago/Turabian StyleLu, Yi, Jing Dai, Na Kong, Jianghuai Liu, Jinkang Gong, and Yuan Yao. 2019. "Internalization Characterization of Si Nanorod with Camouflaged Cell Membrane Proteins Reveals ATXN2 as a Negative Regulator" Cells 8, no. 8: 931. https://doi.org/10.3390/cells8080931