Leucine-Rich Alpha-2-Glycoprotein1 Gene Interferes with Regulation of Apoptosis in Leukemia KASUMI-1 Cells
| Autor: | Hongqian Zhu, Shishan Xiao |
|---|---|
| Rok vydání: | 2018 |
| Předmět: |
0301 basic medicine
Cell Survival Cell Blotting Western Apoptosis Real-Time Polymerase Chain Reaction 03 medical and health sciences 0302 clinical medicine Lab/In Vitro Research Cell Line Tumor Cyclins Gene expression medicine Gene silencing Humans Viability assay Gene Silencing Cell Proliferation Glycoproteins Chemistry Cell Cycle General Medicine Transfection Cell cycle Flow Cytometry Cell biology Leukemia Myeloid Acute 030104 developmental biology medicine.anatomical_structure Cell culture 030220 oncology & carcinogenesis Signal transduction Cell Division Signal Transduction |
| Zdroj: | Medical Science Monitor : International Medical Journal of Experimental and Clinical Research |
| ISSN: | 1643-3750 |
| Popis: | BACKGROUND Leukemia cells have strong proliferation and anti-apoptosis capabilities. The purpose of this study was to investigate the effect of silencing the leucine-rich alpha-2-glycoprotein1 (LRG1) gene, which was found to regulate tumor proliferation and apoptosis in acute myeloid leukemia (AML) cell lines. MATERIAL AND METHODS Plasmid interference technique was used to silence the LRG1 gene in the KASUMI-1 cell line. The cell counting kit-8 (CCK-8) assay was used to test the effect of transduction on cell viability. Cell cycle and apoptosis were detected by flow cytometry. Western blot and quantitative real-time polymerase chain reaction (RT-qPCR) were applied to detect the expression levels of proteins and mRNA, respectively. RESULTS KASUMI-1 cells with the CD34⁺CD38⁻ phenotype were sorted by flow cytometry. After transfection of the siLRG1 plasmid, the level of LRG1 expression was downregulated and cell viability was reduced. Silencing of LRG1 gene blocked KASUMI-1 cells in G0/G1 phase and promoted apoptosis. Further experiments found that LRG1 gene silencing significantly downregulated cell cycle-associated proteins and anti-apoptotic proteins, while upregulating pro-apoptotic proteins. Downregulation of LRG1 gene expression also inhibits signal transduction of the JAK-STAT pathway. CONCLUSIONS LRG1 gene silencing regulates the expression of cyclin and apoptosis-related proteins to reduce cell viability and promote apoptosis, probably through inhibition of the JAK-STAT pathway. |
| Databáze: | OpenAIRE |
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