lncRNA CYTOR靶向miR-139-5p对三阴性乳腺癌细胞增殖的影响及其机制  

Effect of lncRNA CYTOR targeting miR-139-5p on proliferation of triple-negative breast cancer cells and its mechanism

作　　者：董亮亮[1] 徐露露 刘冰[1] 李敏敏 DONG Liangliang;XU Lulu;LIU Bing;LI Minmin(Department of Oncology,Yantai Yuhuangding Hospital,Yantai 264000,China)

机构地区：[1]烟台毓璜顶医院肿瘤内科,山东烟台264000

出　　处：《山东医药》2024年第25期21-25,共5页Shandong Medical Journal

基　　金：山东省医药卫生科技发展计划项目(202103100964);烟台毓璜顶医院科研发展基金(2021-06)。

摘　　要：目的探讨长链非编码RNA(lncRNA)细胞骨架调节因子RNA(CYTOR)靶向微小RNA-139-5p(miR-139-5p)对三阴性乳腺癌细胞增殖的影响及其机制。方法体外传代培养三阴性乳腺癌细胞株HCC1806,取传3代、对数生长期、生长状态良好的细胞进行后续实验。通过starBase数据库预测lncRNA CYTOR与miR-139-5p的结合位点,通过双荧光素酶报告基因实验验证lncRNA CYTOR与miR-139-5p的靶向关系。取HCC1806细胞,随机分为空白对照组、阴性对照组、lncRNA CYTOR沉默组、miR-139-5p过表达组、lncRNA CYTOR沉默+miR-139-5p过表达组。阴性对照组、lncRNA CYTOR沉默组、miR-139-5p过表达组、lncRNA CYTOR沉默+miR-139-5p过表达组分别转染NC mimics、lncRNA CYTOR siRNA、miR-139-5p mimics、lncRNA CYTOR siRNA+miR-139-5p mimics,转染48 h更换新鲜培养基,继续培养24 h。空白对照组常规培养,不予转染。收集各组上述细胞,采用RT-qPCR法检测miR-139-5p、性别决定区Y框蛋白8(SOX8)mRNA表达,采用EdU法检测细胞增殖能力,采用Western blotting法检测ATP结合盒转运蛋白G2(ABCG2)、SOX8表达。结果经starBase数据库预测,lncRNA CYTOR与miR-139-5p存在结合位点;经双荧光素酶报告基因实验验证,lncRNA CYTOR与miR-139-5p存在靶向调控关系。lncRNA CYTOR沉默组、miR-139-5p过表达组、lncRNA CYTOR沉默+miR-139-5p过表达组miR-139-5p相对表达量均高于空白对照组和阴性对照组,SOX8 mRNA相对表达量、EdU阳性细胞比例以及ABCG2、SOX8蛋白相对表达量均低于空白对照组和阴性对照组(P均<0.05);lncRNA CYTOR沉默+miR-139-5p过表达组miR-139-5p相对表达量高于lncRNA CYTOR沉默组和miR-139-5p过表达组,SOX8 mRNA相对表达量、EdU阳性细胞比例以及ABCG2、SOX8蛋白相对表达量均低于lncRNA CYTOR沉默组和miR-139-5p过表达组(P均<0.05);而空白对照组与阴性对照组、lncRNA CYTOR沉默组与miR-139-5p过表达组miR-139-5p、SOX8 mRNA相对表达量以及EdU阳性细胞比例和ABCG2、SOXObjective To investigate the effect of long non-coding RNA(lncRNA)cytoskeleton regulator RNA(CYTOR)targeting microRNA-139-5p(miR-139-5p)on the proliferation of triple-negative breast cancer cells and its mechanism.Methods The triple-negative breast cancer cell line HCC1806 was cultured in vitro,and the cells in the third generation,which were in the logarithmic growth phase and had good growth status,were taken for subsequent experi‑ments.The targeted binding sites of lncRNA CYTOR and miR-139-5p were predicted through the StarBase database,and the targeted relationship between lncRNA CYTOR and miR-139-5p was verified through dual luciferase reporter gene experiments.HCC1806 cells were randomly divided into the blank control group,negative control group,lncRNA CYTOR silencing group,miR-139-5p overexpression group,lncRNA CYTOR silencing+miR-139-5p overexpression group,respec‑tively.Cells in the negative control group,lncRNA CYTOR silencing group,miR-139-5p overexpression group,lncRNA CYTOR silencing+miR-139-5p overexpression group,and lncRNA CYTOR silencing+miR-139-5p overexpression group,were transfected with NC mimics,lncRNA CYTOR siRNA,miR-139-5p mimics,lncRNA CYTOR siRNA+miR-139-5p mimics,respectively.After 48 h of transfection,we replaced with fresh culture medium and continued to culture cells for 24 h.cells in the blank control group were cultured routinely and not transfected.We collected the aforementioned cells from each group.The expression levels of miR-139-5p and SOX8 mRNA were measured using RT-qPCR.The cell prolifer‑ation ability was measured using EdU method.The expression of ATP binding cassette transporter G2(ABCG2)and sexdetermining region Y-box protein 8(SOX8)was measured by Western blotting.Results Through the starBase database prediction,there was a binding site between lncRNA CYTOR and miR-139-5p.The dual luciferase reporter gene experi‑ment confirmed that lncRNA CYTOR had a targeted regulatory relationship with miR-139-5p.The relative expression lev‑els of miR-139-5p in the lncRNA

关 键 词：三阴性乳腺癌 长链非编码RNA细胞骨架调节因子RNA 微小RNA-139-5p 性别决定区Y框蛋白8 ATP结合盒转运蛋白G2 

分 类 号：R737.9[医药卫生—肿瘤]