miR-144-3p/SLC7All轴通过调控铁死亡增强 卵巢癌细胞对顺钳的敏感性.

miR-144-3p/SLC7All axis enhances cisplatin sensitivity in ovarian cancer cells through ferroptosis.

AIM: To explore the effect of microRNA-144-3p (miR-144-3p) on cisplatin resistance in ovarian cancer cells and determine whether its mechanism is related to solute carrier family 7 member 11 (SLC7A11) and ferroptosis. METHODS; After transfecting miR-144-3p mimic into cisplatin-resistant ovarian cancer cell lines, the abundance of miR-144-3p was measured by RT-qPCR. Sensitivity to cisplatin and cell proliferation were determined by CCK-8 and colony-forming assays, respectively. The levels of malondialdehyde(MDA)and glutathione(GSH)were assessed by kit assays. Fe2+ content and reactive oxygen species (ROS) level were detected using Fe2+ probe and ROS fluorescent probe, respectively. Mitochondrial morpghology was observed under transmission electron microscope. Dual-luciferase reporter assay was used to identify and verify the binding between miR-144-3p and SLC7A11. Furthermore, A2780/DDP cancer cell xenograft model was established to evaluate the effect of miR44-3p on tumor growth in vivo. RESULTS: The expression of miR44-3p in drug-resistant cell lines A2780/DDP and SKOV3/DDP was significantly lower than that in parent cell lines A2780 and SKOV3 (PvO. 05). Transfection with miR-144-3p mimic inhibited cell proliferation and enhance the sensitivity of drug-resistant ovarian cancer cells to cisplatin(PV0. 01). Dual-luciferase reporter assay demonstrated that SLC7A11 was a target gene of miR-144-3p, and SLC7A11 overexpression reversed the effect of miR-144-3p on the cell proliferation and cisplatin sensitivity through ferroptosis （P<0. 05）. Additionally, in vivo xenograft experiments showed that miR-144-3p inhibited tumor growth, and down-regulated SLC7A11 and glutathione peroxidase 4 （GPX4）expression（P< 0. 01）, along with the up-regulation of 4-hydroxynonenal（4-HNE）level（P

目的：探讨微小RNA-144-3p(microRNA-144-3p, miR-144-3p)在卵巢癌细胞对顺钳耐药中的作用 并分析其作用机制与溶质载体家族7成员11 (solute carrier family 7 member 11, SLC7A11)和铁死亡是否有关°方 法：将miR-144-3p mimic转染入耐顺钳人卵巢癌细胞株A2780/DDP和SKOV3/DDP后, RT-qPCR法检测miR-144-3p 的表达丰度;CCK-8法检测细胞对顺钳的敏感性;集落形成实验测定细胞增殖情况;试剂盒法评估细胞内丙二醛 (malondialdehyde, MDA)和谷胱甘肽(glutathione, GSH)的水平;Fe"探针及活性氧簇(reactive oxygen species, ROS) 荧光探针检测细胞内含量及ROS水平;透射电子显微镜观察线粒体形态;双萤光素酶报告基因实验验证miR-144-3p与SLC7A11之间的靶向结合。建立耐药细胞株A2780/DDP异种移植瘤模型, 体内 W miR-144-3p对肿瘤生 长的影响。结果：耐药细胞株A2780/DDP和SKOV3/DDP中的miR-144-3p表达显著低于亲本细胞株A2780和 SKOV3(/><0. 05)。转染miR-144-3p mimic可抑制细胞增殖, 增强耐药细胞株对顺钳的敏感性("<0・01)。双萤光素 酶报告基因实验结果显示SLC7A11是miR-144-3p的作用靶点。过表达SLC7A11可通过铁死亡途径逆转miR-144-3p对细胞增殖及化疗敏感性的作用("<0.05)。异种移植瘤实验结果表明miR-144-3p可显著抑制瘤体生长, 抑制 SLC7A11及谷胱甘肽过氧化物酶4(glutathione peroxidase 4, GPX4)蛋白表达("<0. 01), 提高4-規基壬烯醛(4-hy-droxynonenal, 4-HNE)水平(/»<0.01)o结论：miR-144-3p通过靶向SLC7A11而增强耐药卵巢癌细胞对顺钳的敏感 性,其作用机制可能涉及铁死亡过程. [ABSTRACT FROM AUTHOR]

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