miR-17-5p 通过靶向SETD2 调控骨髓增生异常综合征SKM-1 细胞的增 殖与凋亡. (Chinese)

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, pcDNASETD2, 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. [ABSTRACT FROM AUTHOR]

目的：探讨miR-17-5p 和含SET结构域蛋白2（SETD2）对骨髓增生异常综合征（MDS）SKM-1 细胞增殖与凋亡的影响 及其作用机制。方法：收集2019 年3 月至2021 年5 月衡水市人民医院就诊的35 例MDS患者的骨髓标本（MDS组）、35 例健康体 检者的骨髓标本（对照组）, 以及MDS细胞系SKM-1。用qPCR 法检测MDS 骨髓和SKM-1 细胞中miR-17-5p、SETD2 mRNA的 表达水平。双荧光素酶报告基因实验验证miR-17-5p 与SETD2 的靶向关系。利用脂质体转染技术, 分别将si-miR-NC、 si-miR-17-5p、miR-NC、miR-17-5p mimic、pcDNA、pcDNA-SETD2、si-miR-17-5p+si-NC、si-miR-17-5p+si-SETD2 等转染至SKM-1 细胞, CCK-8 法、流式细胞术检测细胞的增殖和凋亡水平, WB法检测细胞中SETD2、C-caspase-3、C-caspase-9 的表达。结果：与 对照组相比, MDS组骨髓中miR-17-5p 表达水平显著升高、SETD2的mRNA和蛋白表达水平均显著降低（均P<0.01）。与si-miR-NC 组相比, si-miR-17-5p 组SKM-1 细胞增殖能力显著降低、凋亡率显著升高, 细胞中C-caspase-3 和C-caspase-9 表达显著升高（均 P<0.01）。miR-17-5p 明显抑制野生型SETD2 细胞的荧光素酶活性（P<0.01）, 并负向调控SETD2 的表达。过表达SETD2 可显著 抑制SKM-1 细胞的增殖并促进细胞凋亡, 同时干扰SETD2 表达则可部分逆转干扰miR-17-5p 对SKM-1 细胞的增殖抑制和凋亡 促进作用。结论：MDS骨髓中miR-17-5p 呈高表达, 干扰miR-17-5p 可抑制SKM-1 细胞增殖并促进细胞凋亡, 其机制与 miR-17-5p 靶向负调控SETD2 的表达有关。. [ABSTRACT FROM AUTHOR]

Copyright of Chinese Journal of Cancer Biotherapy is the property of Editorial Office of Chinese Journal of Cancer Biotherapy and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)