微RNA-25-3p调控PTEN/PI3K/AKT通路影响三阴性乳腺癌细胞增殖、迁移及侵袭的研究OA
Effects of microRNA-25-3p on the proliferation,migration and invasion of TNBC cells by regulating the PTEN/PI3K/AKT signal-ing pathway
目的 探讨微RNA-25-3p(miR-25-3p)通过调控磷酸酶与张力蛋白同源物(PTEN)/磷脂酰肌醇3-激酶(PI3K)/蛋白激酶B(AKT)信号通路对三阴性乳腺癌(TNBC)细胞增殖、侵袭和迁移的影响.方法 选取2025年1月至12月在湖州市妇幼保健院行乳腺癌改良根治术或保乳术的TNBC患者30例,留取患者的TNBC组织及对应癌旁组织标本.采用实时荧光定量聚合酶链反应检测TNBC组织与癌旁组织、MDA-MB-231细胞(TNBC细胞)与MCF-10A细胞(人正常乳腺细胞)miR-25-3p表达水平.体外培养MDA-MB-231 细胞,分别转染miR-25-3p过表达质粒(miR-25-3p mimics)、抑制miR-25-3p表达质粒(miR-25-3p inhibitor)及阴性对照质粒(NC inhibitor).分别采用CCK-8法、细胞划痕实验及Transwell实验评估MDA-MB-231细胞的增殖、迁移及侵袭能力;采用蛋白免疫印迹法检测MDA-MB-231细胞PTEN、PI3K、磷酸化PI3K(p-PI3K)、AKT、磷酸化AKT(p-AKT)蛋白表达.采用双萤光素酶实验验证miR-25-3p与PTEN的靶向关系.结果 TNBC组织miR-25-3p表达水平高于癌旁组织(P＜0.05),MDA-MB-231 细胞miR-25-3p表达水平高于MCF-10A细胞(P＜0.05).抑制miR-25-3p表达后,MDA-MB-231细胞增殖、迁移与侵袭能力均明显降低,PTEN蛋白表达明显上调,p-PI3K、p-AKT蛋白表达均明显下调,差异均有统计学意义(均P＜0.05).过表达miR-25-3p后,MDA-MB-231细胞增殖、迁移与侵袭能力均明显增强,PTEN蛋白表达明显下调,p-PI3K、p-AKT蛋白表达均明显上调,差异均有统计学意义(均P＜0.05).双萤光素酶实验结果表明,miR-25-3p能够靶向并负向调控PTEN的表达.结论 miR-25-3p可能通过靶向抑制PTEN基因的表达,激活PI3K/AKT信号通路,促进TNBC细胞的增殖、迁移和侵袭.
Objective To investigate the effects of microRNA-25-3p(miR-25-3p)on triple-negative breast cancer(TNBC)cell proliferation,invasion,and migration via regulating the phosphatase and tensin homolog(PTEN)/phosphatidylinositol 3-kinase(PI3K)/protein kinase B(AKT)signaling axis.Methods Thirty TNBC patients undergoing modified radical mastectomy or breast-conserving surgery at Huzhou Maternal and Child Health Hospital from January to December 2025 were enrolled.TNBC tissues and adjacent normal tissues of patients were harvested.Quantitative real-time polymerase chain reaction(qRT-PCR)was performed to assess miR-25-3p expression in TNBC tissues and adjacent non-tumor tissues,as well as in MDA-MB-231 cells(TNBC cells)and MCF-10A cells(human normal breast cells).MDA-MB-231 cells were cultured in vitro by being transfected with miR-25-3p mimics,miR-25-3p inhibitor,or negative control(NC).MDA-MB-231 cell proliferation,migration,and invasion were evaluated using CCK-8,wound-healing,and Transwell assays,respectively.Western blotting was employed to detect PTEN,PI3K,phosphorylated PI3K(p-PI3K),AKT,and phosphorylated AKT(p-AKT)protein levels.A dual-luciferase reporter assay was conducted to verify the targeting relationship between PTEN and miR-25-3p.Results miR-25-3p expression was markedly higher in TNBC tissues than in matched adjacent tissues(P＜0.05),and its expression in MDA-MB-231 cells was higher than that in MCF-10A cells(P＜0.05).Knockdown of miR-25-3p substantially suppressed MDA-MB-231 cell proliferation,migration,and invasion,paralleled by upregulated PTEN expression and downregulated p-PI3K and p-AKT expressions(all P＜0.05).In contrast,overexpression of miR-25-3p resulted in enhanced cell proliferation,migration,and invasion of MDA-MB-231,diminished PTEN expression,and increased p-PI3K and p-AKT expressions(all P＜0.05).The dual-luciferase assay demonstrated that miR-25-3p could target and negatively modulate PTEN expression.Conclusion miR-25-3p promotes TNBC cell proliferation,migration,and invasion by targeting and suppressing PTEN gene expression,and activating the PI3K/AKT signaling pathway.
丁秀芳;陈莉杰;姚敏;王晨;邓再兴;张昕;纪存丽;关惠元;彭浦超
313000 湖州市妇幼保健院乳腺科313000 湖州市妇幼保健院乳腺科313000 湖州市妇幼保健院乳腺科浙江省肿瘤医院乳腺外科313000 湖州市妇幼保健院病理科313000 湖州市妇幼保健院病理科313000 湖州市妇幼保健院乳腺科313000 湖州市妇幼保健院乳腺科313000 湖州市妇幼保健院乳腺科
三阴性乳腺癌微RNA磷酸肌醇-3激酶增殖侵袭
Triple-negative breast cancerMicroRNAPhosphatidylinositol 3-kinaseProliferationInvasion
《浙江医学》 2026 (17)
1825-1831,后插1,8
浙江省自然科学基金项目(LY23H160005)浙江省医药卫生科技计划项目(2024KY423)湖州市科技计划项目(2024GYB38)
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