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Overexpression of SHP-1 Enhances the Sensitivity of K562 Cells to Imatinib / 中国实验血液学杂志
Journal of Experimental Hematology ; (6): 46-51, 2016.
Article in Chinese | WPRIM | ID: wpr-272507
ABSTRACT
<p><b>OBJECTIVE</b>To explore the effect of overexpression of SH2-containing tyrosine phosphatase 1 (SHP-1) on sensitivity of chronic myelogenous 1eukemia (CML) K562 cell line to imatinib and its related mechamism.</p><p><b>METHODS</b>K562 cells were infected with the lentiviral plasmids containing the specified retroviral vector (pEX-SHP-1-puro-Lv105) or the mock vector (pEX-EGFP-puro-Lv105). The expression of SHP-1 in K562 cells treated with 0.2 µmol/L imatinib (IM) for 72 h was determined by Western blot. After transfection the CCK-8 assay was used to determine the proliferation of the tramfected K562 cells (K562(SHP-1) and K562(EGFP) cells) at 72 h after exposure to different doses of IM, the half inhibitary concentration (IC50) was calculated. The mechanisms of the overexpression effects of SHP-1 and IM on the proliferation in K562 cells was investigated, the BCR-ABL1 activity and the level of tyrosine phosphorylation of CrkL (pCrkL) was measured by flow cytometry; the Western blot was used to detect the expression and activity of these molecules controlling cell growth, including MAPK, AKT, STAT5 and JAK2.</p><p><b>RESULTS</b>After exposure of K562 cells to 0.08 µmol/L IM for 72 h, there was no significant change of SHP-1 expression in K562 cells. After exposure to 0.2 µmol/L of IM for 72 h, the inhibitory rate of K562(SHP-1) group was higher than that of K562(EGFP) group (P < 0.05), indicating that overexpression of SHP-1 in K562 cells could enhance the proliferation inhtibition effect of IM on K562 cells. The IC50 of IM in K562(SHP-1) cells was the lower as compared with that of K562(EGFP) cells (P < 0.05) after exposure to different concentrations of IM for 72 h. The slope of K562(SHP-1) cells was the largest ranging 0.02 - 0.16 µmol/L of IM. Overexpression of SHP-1 and IM could inhibit the activity BCR-ABL1, MAPK, AKT, STAT5 and JAK2 signaling pathways in the K562 cell line and displayed a synergistic effect.</p><p><b>CONCLUSION</b>SHP-1 inhibits BCR-ABL1, MAPK, AKT, STAT5 and JAK2 signaling pathways in K562 cells, the overexpression of SHP-1 can enhance the sensitivity of K562 cells to IM.</p>
Subject(s)
Full text: Available Index: WPRIM (Western Pacific) Main subject: Pharmacology / Phosphorylation / Transfection / Signal Transduction / Drug Resistance, Neoplasm / K562 Cells / Cell Proliferation / Protein Tyrosine Phosphatase, Non-Receptor Type 6 / Imatinib Mesylate / Genetic Vectors Limits: Humans Language: Chinese Journal: Journal of Experimental Hematology Year: 2016 Type: Article

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Full text: Available Index: WPRIM (Western Pacific) Main subject: Pharmacology / Phosphorylation / Transfection / Signal Transduction / Drug Resistance, Neoplasm / K562 Cells / Cell Proliferation / Protein Tyrosine Phosphatase, Non-Receptor Type 6 / Imatinib Mesylate / Genetic Vectors Limits: Humans Language: Chinese Journal: Journal of Experimental Hematology Year: 2016 Type: Article