Long non-coding RNA VPS9D1-AS1对乳腺癌细胞恶性行为的影响和机制  

Effect of long non-coding RNA VPS9D1-AS1 on malignant behavior of breast cancer cells and its mechanism

作　　者：侯红 冯姗姗 孙华静 盖磊 方堃 HOU Hong;FENG Shanshan;SUN Huajing;GAI Lei;FANG Kun(Department of Breast Surgery,Qingdao Central Hospital,University of Health and Rehabilitation Sciences,Qingdao 266042,Shandong,China;Department of Internal Medicine,Boxing County Traditional Chinese Medicine Hospital,Boxing 256599,Shandong,China)

机构地区：[1]康复大学青岛中心医院乳腺外科,山东青岛266042 [2]山东省博兴县中医医院内科,山东博兴256599

出　　处：《贵州医科大学学报》2024年第10期1490-1497,共8页Journal of Guizhou Medical University

基　　金：山东省中医药科技项目(Q-2022044)。

摘　　要：目的探讨长链非编码RNA(LncRNA)VPS9D1-AS1对乳腺癌细胞恶性行为的影响和机制。方法收集49例接受手术的乳腺癌患者的乳腺癌组织和癌旁组织,体外培养人正常乳腺上皮细胞MCF-10A和乳腺癌细胞系MCF-7、T47D、BT549,采用实时荧光定量PCR(RT-qPCR)检测miR-4695-5p及LncRNA VPS9D1-AS1表达水平;选择MCF-7细胞分别转染si-NC、si-LncRNA VPS9D1-AS1、pcDNA、pcDNA-LncRNA VPS9D1-AS1、miR-NC、miR-4695-5p模拟物、anti-miR-NC+si-LncRNA VPS9D1-AS1、anti-miR-4695-5p+si-LncRNA VPS9D1-AS1,采用RT-qPCR检测MCF-7细胞中LncRNA VPS9D1-AS1、miR-4695-5p表达水平,CCK-8和Transwell实验评估细胞存活率和迁移侵袭数,蛋白质印记法检测IL-6、MMP2、MMP9蛋白水平;双荧光素酶实验证实LncRNA VPS9D1-AS1与miR-4695-5p的靶向关系。结果LncRNA VPS9D1-AS1在乳腺癌组织和MCF-7、T47D、BT549细胞系中表达上调(P<0.05),miR-4695-5p表达下调(P<0.05),MCF-7细胞中LncRNA VPS9D1-AS1、miR-4695-5p变化最明显,选择MCF-7细胞进行后续实验;转染LncRNA VPS9D1-AS1后,MCF-7细胞存活率、细胞迁移和侵袭数以及IL-6、MMP2、MMP9蛋白水平降低(P<0.05);转染miR-4695-5p模拟物后,miR-4695-5p表达升高(P<0.05),MCF-7细胞存活率、细胞迁移和侵袭数及IL-6、MMP2、MMP9蛋白水平降低(P<0.05);转染pcDNA-LncRNA VPS9D1-AS1后,MCF-7细胞中miR-4695-5p表达下降(P<0.05);共转染anti-miR-4695-5p+si-LncRNA VPS9D1-AS1后,MCF-7细胞存活率、迁移数和侵袭数及IL-6、MMP2、MMP9蛋白水平升高(P<0.05)。结论干扰LncRNA VPS9D1-AS1通过靶向上调miR-4695-5p抑制促炎因子IL-6表达,进而抑制乳腺癌MCF-7细胞增殖、迁移及侵袭。Objective To explore the effect of long non-coding RNA(LncRNA)VPS9D1-AS1 on malignant behavior of breast cancer cells and its mechanism.Methods Breast cancer tissues and adjacent tissues were collected from 49 patients with surgically resected breast cancer.Human normal mammary epithelial cell line MCF-10A and breast cancer cell lines MCF-7,T47D,and BT549 were cultured in vitro.Real time quantitative PCR(RT-qPCR)was used to detect the expression levels of miR-4695-5p and LncRNA VPS9D1-AS1.MCF-7 cells were transfected with si-NC,si-LncRNA VPS9D1-AS1,pcDNA,PCDNA-LncRNA VPS9D1-AS1,miR-NC,miR-4695-5p mimics,anti-miR-NC+si-LncRNA VPS9D1-AS1 and anti-miR-4695-5p+si-LncRNA VPS9D1-AS1,respectively.RT-qPCR was applied to detect the expressions of LncRNA VPS9D1-AS1 and miR-4695-5p in MCF-7 cells.CCK-8 assay and Transwell assay were used to examine cell survival rate,cell numbers of migration and invasion.Western blot was applied to detect the protein levels of interleukin 6(IL-6),matrix metalloproteinase 2(MMP2),and MMP9.Dual luciferase reporter assay was confirmed the interaction between LncRNA VPS9D1-AS1 and miR-4695-5p.Results LncRNA VPS9D1-AS1 was upregulated in breast cancer tissues,MCF-7,T47D,and BT549 cell lines(P<0.05),while miR-4695-5p was downregulated(P<0.05).LncRNA VPS9D1-AS1 and miR-4695-5p exhibited the most obvious changes in MCF-7 cells.MCF-7 cells were selected for follow-up experiments.After transfection with LncRNA VPS9D1-AS1,MCF-7 cell survival rate,the numbers of cell migration and invasion and the protein levels of IL-6,MMP2,and MMP9 were decreased(P<0.05).After transfection with miR-4695-5p mimics,MCF-7 cell survival rate,the numbers of cell migration and invasion and the protein levels of IL-6,MMP2 and MMP9 were decreased(P<0.05),while miR-4695-5p expression was increased(P<0.05).After transfection with pcDNA-LncRNA VPS9D1-AS1,miR-4695-5p expression in MCF-7 cells was decreased(P<0.05).After co-transfection with anti-miR-4695-5p+si-LncRNA VPS9D1-AS1,MCF-7cell survival rate,the numbers and invasion n

关 键 词：乳腺肿瘤 长链非编码RNA LncRNA VPS9D1-AS1高表达 上皮细胞MCF-10A 增殖 迁移 侵袭 

分 类 号：R737.9[医药卫生—肿瘤]