Zhongguo shi yan xue ye xue za zhi · 2020
[Function of MEIS1 and miR-425 to the Regulating of Chronic Myeloid Leukemia Cell Proliferation].
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Zhao Xu-Hong, Yang Gui-Hua, Li Dan-Dan
Аннотация
OBJECTIVE: To investigate the function and mechanism of transcription factor of MEIS1 and miR-425 to the proliferation of chronic myeloid leukemia cell K562.
METHODS: Bioinformatic prediction was used to analyze the binding of MEIS1 in miR-425 promoter region. ChIP-qPCR coupled with dual luciferase assay was used to detect the combination of MEIS1 and the transcription activity of miR-425, and its regulative role in the transcription activity miR-425. CCK-8 was used to detect the effect of MEIS1 and miR-425 on cell proliferation. Flow cytometry with PI staining was used to detected the effect of MEIS1 and miR-425 on K562 cell cycle progression. Western blot was used to examine the effect of miR-452 on the expression level of MEIS1.
RESULTS: MEIS1 could bind the promoter of miR-425 and repressed its transcription. After K562 was transfected by shRNA, the K562 cell proliferation and cell cycle progression was significantly inhibitied. Moreover, after K562 cells were transfected by miR-425 mimic, cell proliferation and cell cycle was inhibited. The expression level of MEIS1 could be inhibited by the combination of miR-425 and MEIS1 3'UTR.
CONCLUSION: MEIS1 can inhibit the activity of miR-425 in transcriptional level, while the miR-425 can suppress the expression of MEIS1 protein in post-transnational level. Therefore, a regulatory circuit comprising from MEIS1 and miR-425 regulates K562 cell proliferation.
METHODS: Bioinformatic prediction was used to analyze the binding of MEIS1 in miR-425 promoter region. ChIP-qPCR coupled with dual luciferase assay was used to detect the combination of MEIS1 and the transcription activity of miR-425, and its regulative role in the transcription activity miR-425. CCK-8 was used to detect the effect of MEIS1 and miR-425 on cell proliferation. Flow cytometry with PI staining was used to detected the effect of MEIS1 and miR-425 on K562 cell cycle progression. Western blot was used to examine the effect of miR-452 on the expression level of MEIS1.
RESULTS: MEIS1 could bind the promoter of miR-425 and repressed its transcription. After K562 was transfected by shRNA, the K562 cell proliferation and cell cycle progression was significantly inhibitied. Moreover, after K562 cells were transfected by miR-425 mimic, cell proliferation and cell cycle was inhibited. The expression level of MEIS1 could be inhibited by the combination of miR-425 and MEIS1 3'UTR.
CONCLUSION: MEIS1 can inhibit the activity of miR-425 in transcriptional level, while the miR-425 can suppress the expression of MEIS1 protein in post-transnational level. Therefore, a regulatory circuit comprising from MEIS1 and miR-425 regulates K562 cell proliferation.