敲低 LINC00665 通过竞争性结合 miR-3619-5p 和抑制连环蛋白 beta 1 抑制乳腺癌的增殖和侵袭。
Knockdown of LINC00665 inhibits proliferation and invasion of breast cancer via competitive binding of miR-3619-5p and inhibition of catenin beta 1.
机构信息
Department of Breast Surgery, The Affiliated Cancer Hospital of Zhengzhou University, Zhengzhou, Henan P.R. China.
Department of Medical Oncology, The Affiliated Cancer Hospital of Zhengzhou University, No. 127, Dongming Road, Jinshui District, Zhengzhou, 450008 Henan P.R. China.
出版信息
Cell Mol Biol Lett. 2020 Sep 24;25:43. doi: 10.1186/s11658-020-00235-8. eCollection 2020.
BACKGROUND
Long intergenic non-protein coding RNA00665 (LINC00665) plays a crucial tumorigenic role in many cancers, such as gastric cancer and lung adenocarcinoma. However, its role and mechanism of action in the progression of breast cancer (BC) are unknown.
METHODS
LINC00665 expression levels were determined using quantitative polymerase chain reaction analysis with BC tissues and cell lines. BC cell proliferation was tested by performing 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assays, whereas BC cell migration and invasion capabilities were analyzed by performing transwell migration assays. Percentages of apoptotic cells were measured by flow cytometry. Interactions between LINC00665 and miR-3169-5p were examined by performing luciferase reporter assays, and the expression levels of proteins, such as β-catenin, were examined by western blot analysis.
RESULTS
LINC00665 was expressed at high levels in BC tissues and cells. Upregulated LINC00665 expression correlated with tumor size and tumor, node, and metastasis stages, but not with the age of patients. LINC00665 knockdown inhibited BC cell proliferation, migration, and invasion, whereas it promoted apoptosis. Moreover, bioinformatics analysis and the luciferase reporter assay revealed that LINC00665 bound the microRNA (miR) miR-3619-5p. miR-3619-5p expression correlated negatively with LINC00665 expression in BC tissues. miR-3619-5p overexpression inhibited BC cell proliferation, migration, and invasion, but promoted apoptosis. Simultaneous knockdown of LINC00665 and miR-3619-5p led to increased cell proliferation, migration, and invasion, and inhibited apoptosis. Additionally, catenin beta 1, which encodes the β-catenin protein, was the target gene of miR-3619-5p. β-catenin expression clearly decreased after LINC00665 knockdown and miR-3619-5p overexpression, but increased after simultaneous knockdown of LINC00665 and miR-3619-5p.
CONCLUSION
LINC00665 knockdown inhibited BC cell proliferation and invasion by binding miR-3619-5p and inhibiting β-catenin expression.
背景
长链非编码 RNA00665（LINC00665）在许多癌症中发挥着关键的致癌作用，如胃癌和肺腺癌。然而，其在乳腺癌（BC）进展中的作用和机制尚不清楚。
方法
采用定量聚合酶链反应分析检测 BC 组织和细胞系中的 LINC00665 表达水平。通过 3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四唑溴盐（MTT）实验检测 BC 细胞增殖，通过 Transwell 迁移实验分析 BC 细胞迁移和侵袭能力。通过流式细胞术测量细胞凋亡的百分比。通过荧光素酶报告实验检测 LINC00665 与 miR-3169-5p 之间的相互作用，通过 Western blot 分析检测 β-连环蛋白等蛋白的表达水平。
结果
LINC00665 在 BC 组织和细胞中高表达。上调的 LINC00665 表达与肿瘤大小和肿瘤、淋巴结和转移分期相关，但与患者年龄无关。LINC00665 敲低抑制 BC 细胞增殖、迁移和侵袭，而促进细胞凋亡。此外，生物信息学分析和荧光素酶报告实验显示，LINC00665 与 microRNA（miR）miR-3619-5p 结合。BC 组织中 LINC00665 的表达与 miR-3619-5p 的表达呈负相关。miR-3619-5p 过表达抑制 BC 细胞增殖、迁移和侵袭，但促进细胞凋亡。同时敲低 LINC00665 和 miR-3619-5p 导致细胞增殖、迁移和侵袭增加，凋亡减少。此外，编码 β-连环蛋白蛋白的 catenin beta 1 是 miR-3619-5p 的靶基因。LINC00665 敲低和 miR-3619-5p 过表达后，β-连环蛋白表达明显降低，但同时敲低 LINC00665 和 miR-3619-5p 后，β-连环蛋白表达增加。
结论
LINC00665 通过与 miR-3619-5p 结合抑制 β-连环蛋白表达，从而抑制 BC 细胞增殖和侵袭。
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