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miR-17-5p regulates the proliferation and apoptosis of myelodysplastic syndrome SKM-1 cells by targeting SETD2
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Abstract:
Objective: To investigate the effect of miR-17-5p and SET domain containing 2 (SETD2) on proliferation and apoptosis of myelodysplastic syndrome (MDS) SKM-1 cells and its mechanism. Methods: Bone marrow samples of 35 MDS patients (MDS group) and 35 healthy persons (control group) who had treatment or health checkup in Hengshui People's Hospital from March 2019 to May 2021 were collected; in addition, MDS cell line SKM-1 was also collected for this study. The mRNA expression levels of miR-17-5p and SETD2 in MDS bone marrow and SKM-1 cells were detected by qPCR. The targeting relationship between miR-17-5p and SETD2 was verified using dual-luciferase reporter gene assay. si-miR-NC, si-miR-17-5p, miR-NC, miR-17-5p mimics, pcDNA, pcDNA[1]SETD2, si-miR-17-5p+si-NC, and si-miR-17-5p+si-SETD2 were respectively transfected into SKM-1 cells using liposome transfection technology. CCK-8 method and flow cytometry were used to detect proliferation and apoptosis of SKM-1 cells, and WB method was used to detect the expression of SETD2, C-caspase-3 and C-caspase-9. Results: Compared with the control group, the expression level of miR-17-5p in bone marrow of MDS group significantly elevated, while the mRNA and protein expression levels of SETD2 significantly decreased (all P<0.01). Compared with si-miR-NC group, the proliferation ability of SKM-1 cells in si-miR-17-5p group decreased significantly, while the apoptosis rate and the expression of C-caspase-3 and C-caspase-9 increased significantly (all P<0.01).miR-17-5p significantly inhibited the luciferase activity of the cells with wild-type SETD2 (P<0.01), and negatively regulated the expression of SETD2. Overexpression of SETD2 significantly inhibited the proliferation and promoted apoptosis of SKM-1 cells, while simultaneously interfering with the expression of SETD2 partially reversed the proliferation inhibition and apoptosis promotion effect of miR-17-5p knockdown on SKM-1 cells. Conclusion: miR-17-5p is highly expressed in MDS bone marrow. Knockdown of miR-17-5p can inhibit proliferation and promote apoptosis of SKM-1 cells, the mechanism of which may be related to the negative regulation of SETD2 expression by miR-17-5p.
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