circRNA_MYLK靶向miR-92a-3p调控急性B淋巴细胞白血病细胞增殖及凋亡

doi:10.3969/j.issn.1006-6233.2023.07.05

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摘要 目的: 探讨环状RNA_MYLK(circRNA_MYLK)对急性淋巴细胞白血病细胞增殖及凋亡的影响及其可能作用机制。方法: 选取四川大学华西医院收治的56例急性淋巴细胞白血病患者为研究组,同时选取45例健康志愿者为对照组。取两组骨髓样本后分离单个核细胞,并通过流式细胞仪分选B淋巴细胞;qRT-PCR法检测单个核细胞中circRNA_MYLK、微小RNA-92a-3p(miR-92a-3p)表达量;Pearson法分析急性淋巴细胞白血病患者单个核细胞中circRNA_MYLK与miR-92a-3p相关性。体外培养人急性淋巴细胞白血病细胞系(SUP-B15、NALM-6、BALL-1)和正常B淋巴细胞,并将si-NC、si-circRNA_MYLK、anti-miR-NC、anti-miR-92a-3p、miR-NC、miR-92a-3pmimic、si-circRNA_MYLK和anti-miR-NC、si-circRNA_MYLK和anti-miR-92a-3p转染至SUP-B15细胞;qRT-PCR法检测circRNA_MYLK、miR-92a-3p表达量;CCK-8检测细胞增殖能力;流式细胞仪检测细胞周期和凋亡;双荧光素酶报告实验验证circRNA_MYLK与miR-92a-3p靶向作用。结果: 与对照组和正常B淋巴细胞比较,研究组患者单个核细胞和人急性淋巴细胞白血病细胞系(SUP-B15、NALM-6、BALL-1)中circRNA_MYLK表达量升高(P<0.05),miR-92a-3p表达量降低(P<0.05)。急性淋巴细胞白血病患者单个核细胞中circRNA_MYLK与miR-92a-3p呈负相关(r=-0.9682)。干扰circRNA_MYLK或过表达miR-92a-3p降低SUP-B15细胞存活率和S期细胞比例(P<0.05),增加G₀/G₁期细胞比例、细胞凋亡率(P<0.05);circRNA_MYLK可靶向调节miR-92a-3p的表达;抑制miR-92a-3p表达可逆转干扰circRNA_MYLK表达对SUP-B15细胞增殖、细胞周期、凋亡的作用(P<0.05)。结论: 干扰circRNA_MYLK表达可诱导急性淋巴细胞白血病细胞凋亡并抑制细胞增殖,其机制与靶向调控miR-92a-3p表达有关。

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	饶琦
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关键词 ：急性淋巴细胞白血病, circRNA_MYLK, miR-92a-3p, 细胞增殖, 凋亡

Abstract：Objective: To explore the effect of circRNA_MYLK on the proliferation and apoptosis of acute lymphoblastic leukemia cells and its possible mechanism. Methods: The 56 patients with acute lymphoblastic leukemia admitted to West China Hospital of Sichuan University were selected as the research group,and 45 healthy volunteers were selected as the control group.Mononuclear cells were isolated from bone marrow samples from two groups,and B lymphocytes were sorted by flow cytometry.The expression levels of circRNA_MYLK and microRNA-92a-3p (miR-92a-3p) in mononuclear cells were detected by qRT-PCR; the correlation between the expression of circRNA_MYLK and miR-92a-3p in the mononuclear cells of patients with acute lymphoblastic leukemia was analyzed by Pearson method.Human acute lymphoblastic leukemia cell lines (SUP-B15,NALM-6,BALL-1) and normal B lymphocytes were cultured in vitro and transfected into SUP-B15 cells with si-NC,si-circRNA_MYLK,anti-miR-NC,anti-miR-92a-3p,miR-NC,miR-92a-3pmimic,si-circRNA_MYLK and anti-miR-NC,si-circRNA_MYLK and anti-miR-92a-3p,respectively.CCK-8 experiment was used to detect cell proliferation ability; flow cytometry was used to detect the cell cycle and apoptosis; dual luciferase reporter assay verified the targeting effect of circRNA_MYLK and miR-92a-3p. Results: Compared with the control group and normal B lymphocytes,the expression of circRNA_MYLK in the mononuclear cells of the patients in the research group and human acute lymphoblastic leukemia cell lines (SUP-B15,NALM-6,BALL-1) was increased (P<0.05),while the expression of miR-92a-3p was decreased (P<0.05).circRNA_MYLK was negatively correlated with miR-92a-3p in mononuclear cells of patients with acute lymphoblastic leukemia (r=-0.9682).Interfering with circRNA_MYLK or overexpression of miR-92a-3p decreased the cell survival rate of SUP-B15 cells and the proportion of cells in S phase (P<0.05),and increased the proportion of cells in G₀/G₁ phase and apoptosis rate (P<0.05); circRNA_MYLK could target and regulate the expression of miR-92a-3p; inhibition of miR-92a-3p expression reversed the effect of interfering circRNA_MYLK expression on SUP-B15 cell proliferation,cell cycle and apoptosis (P<0.05). Conclusion: Interfering with the expression of circRNA_MYLK could induce apoptosis of acute lymphoblastic leukemia cells and inhibit cell proliferation,and its mechanism is related to targeted regulation of miR-92a-3p expression.

Key words： Acute lymphocytic leukemia circRNA_MYLK miR-92a-3p Cell proliferation Apoptosis

基金资助:四川省自贡市重点科技计划项目,(编号:2017SF13)

引用本文:

饶琦, 王丹丹, 罗婷. circRNA_MYLK靶向miR-92a-3p调控急性B淋巴细胞白血病细胞增殖及凋亡[J]. 河北医学, 2023, 29(7): 1080-1087.
RAO Qi, WANG Dandan, LUO Ting. circRNA_MYLK Regulating the Proliferation and Apoptosis of Acute Lymphoblastic Leukemia Cells by Targeting MiR-92a-3p. HeBei Med, 2023, 29(7): 1080-1087.

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http://www.hbyxzzs.cn/CN/10.3969/j.issn.1006-6233.2023.07.05 或 http://www.hbyxzzs.cn/CN/Y2023/V29/I7/1080

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