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| Status |
Public on Jan 17, 2024 |
| Title |
Nucleos(t)ide analogs for hepatitis B virus infection differentially regulate the growth factor signaling in hepatocytes |
| Organism |
Homo sapiens |
| Experiment type |
Protein profiling by protein array
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| Summary |
Recent clinical studies have suggested that the risk of developing hepatocellular carcinoma might be lower in chronic hepatitis B (CHB) patients receiving tenofovir disoproxil fumarate (TDF) than in patients receiving entecavir (ETV), although there is no difference in biochemical and virological remission between two drugs. The effects of nucleoside analogs (NsAs; lamivudine and ETV) or nucleotide analogs (NtAs; adefovir disoproxil [ADV], TDF, and tenofovir alafenamide [TAF]) on cell growth and the expression of growth signaling molecules were investigated. The tumor inhibitory effects of NsAs or NtAs were evaluated using a mouse xenograft model, and protein phosphorylation profiles were investigated.
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| Overall design |
1,373 signal transduction proteins with a glutathione S-transferase-tag were synthesized and loaded onto a glutathione-coated glass slide. Every protein was spotted (approximately 500 nL per spot) six times (n = 6) per slide, while ensuring they did not denature or dry. During spotting, humidity was maintained at 40%–60%, and the wetness of each drop was confirmed by microscopic observation of the liquid droplets. Cell lysates of 100 μg total protein were applied to the array with additional ATP at 30°C for 3 h. After termination of the kinase reaction, the array was washed with Tris-buffered saline containing 0.05% Tween 20 and stained with a 4G10 phosphorylated protein-specific antibody (Merck, #05-1050) and a secondary fluorescein-conjugated antibody (Thermo, #A21235) to detect the phosphorylated tyrosine residues. Signal data were averaged (n=6), filtered and normalized by BRB-ArrayTools (https://brb.nci.nih.gov/BRB-ArrayTools).
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| Contributor(s) |
Shimizu R, Murai K, Kagiwada H, Horimoto K, Honda M |
| Citation(s) |
38180972 |
| Submission date |
Dec 11, 2023 |
| Last update date |
Jan 17, 2024 |
| Contact name |
Masao Honda |
| E-mail(s) |
mhondag@gmail.com
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| Phone |
0762652243
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| Organization name |
Kanazawa University
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| Department |
Gastroenterology
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| Street address |
Takara-Machi 13-1
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| City |
Kanazawa |
| ZIP/Postal code |
920-8641 |
| Country |
Japan |
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| Platforms (1) |
| GPL34000 |
Protein array for phosphorylation measurement and pathway analysis |
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| Samples (8)
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| Relations |
| BioProject |
PRJNA1051040 |
| Supplementary file |
Size |
Download |
File type/resource |
| GSE249861_raw_data.txt.gz |
24.4 Kb |
(ftp)(http) |
TXT |
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