circ_0061140靶向miR-338-3p调控多发性骨髓瘤细胞的增殖和凋亡

刘玉香; 江庭秀; 汤杰

doi:10.13898/j.cnki.issn.1000-2200.2024.04.007

circ_0061140靶向miR-338-3p调控多发性骨髓瘤细胞的增殖和凋亡

circ_0061140 regulates the proliferation and apoptosis of multiple myeloma cells through targeting miR-338-3p

摘要

摘要:
目的研究环状RNA 0061140(circ_0061140)对多发性骨髓瘤Sko-007细胞增殖和凋亡的影响和调控机制。
方法采用生物信息学数据库预测circ_0061140的靶miRNA，双荧光素酶报告实验证实circ_0061140与miR-338-3p靶向关系。实时定量PCR分析健康对照者和多发性骨髓瘤病人外周血中circ_0061140和miR-338-3p表达。将circ_0061140小干扰RNA(si-circ_0061140)、miR-338-3p模拟物、si-circ_0061140联合miR-338-3p抑制剂分别转染Sko-007，采用CCK-8法、流式细胞术分析干扰circ_0061140或过表达miR-338-3p或同时抑制circ_0061140和miR-338-3p对Sko-007细胞增殖、凋亡的影响。蛋白质印迹法分析B细胞淋巴瘤-2蛋白(Bcl-2)和Bcl相关x蛋白(Bax)表达变化。
结果 miR-338-3p是circ_0061140达靶基因。circ_0061140在多发性骨髓瘤病人血清中的表达显著高于健康对照者(P < 0.01)，miR-338-3p在多发性骨髓瘤病人血清中的表达显著低于健康对照者(P < 0.01)。干扰circ_0061140表达后Sko-007细胞增殖活力、Bcl-2表达显著降低(P < 0.01)，miR-338-3p和Bax表达、细胞凋亡率显著升高(P < 0.01)。过表达miR-338-3p后Sko-007细胞增殖活力、Bcl-2表达显著降低(P < 0.01)，细胞凋亡率、Bax表达显著升高(P < 0.01)。与干扰circ_0061140比较，同时抑制circ_0061140和miR-338-3p表达后Sko-007细胞增殖活力、Bcl-2表达显著升高(P < 0.01)，细胞凋亡率、Bax表达显著降低(P < 0.01)。
结论 circ_0061140通过靶向miR-338-3p促进多发性骨髓瘤细胞增殖，抑制细胞凋亡，在多发性骨髓瘤中具有致癌作用。

Abstract:
Objective To study the effect and regulatory mechanism of circular RNA 0061140 (circ_0061140) on the proliferation and apoptosis of multiple myeloma Sko-007 cells.
Methods The bioinformatics database was used to predict the target miRNA of circ_0061140, and the dual luciferase report experiment was applied to confirm the targeting relationship between circ_0061140 and miR-338-3p.Expression of circ_0061140 and miR-338-3p in the peripheral blood of healthy controls and multiple myeloma patients was analyzed by real-time quantitative PCR.Circ_0061140 small interfering RNA (si-circ_0061140), miR-338-3p mimic, si-circ_0061140 combined with miR-338-3p inhibitor were transfected into Sko-007 cells respectively.The effects of interference with circ_0061140 or miR-338-3p overexpression or inhibition of circ_0061140 and miR-338-3p on Sko-007 cells proliferation and apoptosis were analyzed by CCK-8 method and flow cytometry.Expression of B-cell lymphoma-2 protein (Bcl-2) and Bcl-related x protein (Bax) was detected using Western blotting.
Results MiR-338-3p was the target gene of circ_0061140.The expression of circ_0061140 was significantly increased (P < 0.01), while the the expression of miR-338-3p was significantly reduced (P < 0.01) in in the serum of patients with multiple myeloma compared with healthy controls.The proliferation activity and Bcl-2 expression of Sko-007 cells were significantly reduced (P < 0.01), and the expression of miR-338-3p and Bax, as well as cell apoptosis rate were significantly increased after interfering with circ_0061140 (P < 0.01).The proliferation activity and Bcl-2 expression of Sko-007 cells were significantly reduced (P < 0.01), and the apoptosis rate and Bax expression were significantly increased after overexpression of miR-338-3p (P < 0.01).Compared with interference circ_0061140, cell proliferation activity and Bcl-2 expression of Sko-007 were significantly increased (P < 0.01), and the apoptosis rate and Bax expression were significantly reduced after simultaneously inhibiting the expression of circ_0061140 and miR-338-3p (P < 0.01).
Conclusions Circ_0061140 acts as an oncogene to promote the proliferation and inhibit the apoptosis of multiple myeloma cells through targeting miR-338-3p.

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