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Effect of Stably Down-regulating FMI Expression of K562 Cells on Sensitivity of K562 cells to Imatinib Mesylate / 中国实验血液学杂志
Journal of Experimental Hematology ; (6): 708-716, 2019.
Article in Chinese | WPRIM | ID: wpr-771896
ABSTRACT
OBJECTIVE@#To investigate the effect of stably down-regulating the FMI expression of K562 cells on the sensitivity of K562 cells to Imatinib (IM) and its possible mechanism.@*METHODS@#Western-blot was used to detect the expression of FMI protein in K562 cells and peripheral blood mononuclear cells from the patients with chronic myelogenous leukemia, chronic myeloid blast crisis and healthy volunteers. The specific interference sequences targeting at the human FMI gene were designed and ligated into the lentiviral vector LV3; the three plasmid system-packaged lentivirus particles were used to transfect K562 cells to screen K562 cells that stably down-regulated FMI. CCK-8 assay and flow cytometry were used to determine effect of IM on cell proliferation and apoptosis. The transcription level of FMI and Fz8 in leukemia cells was detected by fluorescent quantitative PCR. The protein expression levels of FMI, Fz8, NFAT1, BCR-ABL and β-catenin in leukemia cells were detected by Western-blot.@*RESULTS@#The expression of FMI protein could be detected in peripheral blood mononuclear cells of the patients with CML-BC and K562 cells, the FMI expression could not be detected in all the patients with CML-CP and healthy volunteers. The recombinant lentiviral vector LV3/FMI had been successfully constructed the lentivirus was packaged, and the K562 cells stably down-regulating the FMI protein were screened. After stable down-regulation of FMI expression in K562 cells, the proliferation rate of leukemia cells decreased and the apoptosis rate was increased under the same drug concentration. Both the transcription and protein expression levels of Fz8 decreased. The NFAT1 total protein level increased, as well as the nuclear translocation of protein was enhanced. There was no significant change in the expression level of BCR-ABL fusion protein. The expression level of β-catenin protein decreased.@*CONCLUSION@#After the stable down-regulation of FMI expression, the sensitivity of K562 cells to IM and apoptosis of cells increase, which are performed possibly by inhibiting the FMI-Fz8 signaling pathway and activating the Ca-NFAT and Wnt/β-catenin signaling pathway.
Subject(s)
Full text: Available Index: WPRIM (Western Pacific) Main subject: Leukocytes, Mononuclear / Leukemia, Myelogenous, Chronic, BCR-ABL Positive / Fusion Proteins, bcr-abl / Apoptosis / Drug Resistance, Neoplasm / K562 Cells / Imatinib Mesylate Type of study: Diagnostic study Limits: Humans Language: Chinese Journal: Journal of Experimental Hematology Year: 2019 Type: Article

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Full text: Available Index: WPRIM (Western Pacific) Main subject: Leukocytes, Mononuclear / Leukemia, Myelogenous, Chronic, BCR-ABL Positive / Fusion Proteins, bcr-abl / Apoptosis / Drug Resistance, Neoplasm / K562 Cells / Imatinib Mesylate Type of study: Diagnostic study Limits: Humans Language: Chinese Journal: Journal of Experimental Hematology Year: 2019 Type: Article