竞争性内源性RNA(ceRNA)的调控网络广泛存在于肿瘤中,影响肿瘤相关基因的表达,从而在人类肿瘤的发生发展和预后中发挥重要作用。在这项研究中,我们探讨了LINC00665作为ceRNA在乳腺癌中的作用和机制。我们利用生物信息学分析了LINC00665在乳腺癌中的表达和作用靶点,并通过CCK8检测它们对乳腺癌细胞的影响,集落形成和流式细胞术测定。从我们的结果来看,LINC00665敲除抑制增殖,MCF7和MDA-MB-231细胞通过激活AKT/mTOR信号通路诱导细胞凋亡和迁移侵袭。LINC00665有五个潜在的下游靶miRNA(miR-542-3p,miR-624-5p,miR-641,miR-425-5p,和miR-30-3p)。在双荧光素酶报告基因测定中,转染miR-641模拟物的细胞的荧光活性降低,敲低LINC00665后miR-641的表达显著下降。miR-641模拟物显著抑制MCF7和MDA-MB-231细胞的增殖和侵袭。我们使用qPCR检测了miR-641的五个潜在直接靶标(SRCAP,SIKE1NADK,KHDC4和HSPG2)。SRCAP在miR-641过表达细胞中的表达显着下降,并且通过双荧光素酶报告基因测定进一步证实了SRCAP的3UTR和miR-641的结合。SRCAP阻断miR-641或si-LINC00665在MCF7和MDA-MB-231细胞中诱导的增殖和侵袭抑制。总之,LINC00665可以通过构建miR-641和靶向SRCAP促进乳腺癌细胞的存活和转移。该研究为人类乳腺癌的靶向治疗提供了新的潜在靶点。
The regulatory network of competing endogenous RNAs (ceRNA) exists widely in tumors and affects the expression of cancer-related genes, thus playing an important role in the development and prognosis of human tumors. In this research, we explored the role and mechanism of LINC00665 as a ceRNA in breast cancer. We analyzed the expression and targets of LINC00665 in breast cancer using bioinformatics, and detected their effects on breast cancer cells by CCK8, transwell, colony formation and flow cytometry assays. From our results, LINC00665 knockdown suppressed the proliferation, migration and invasion and induced the apoptosis through inactivating the AKT/mTOR signaling pathway in MCF7 and MDA-MB-231 cells. LINC00665 had five potential downstream target miRNAs (miR-542-3p, miR-624-5p, miR-641, miR-425-5p, and miR-30-3p). In dual-luciferase report gene assay, the fluorescence activity of cells transfected with miR-641 mimics decreased, and the expression of miR-641 decreased significantly after knocking down LINC00665. miR-641 mimics significantly inhibited cell proliferation and invasion in MCF7 and MDA-MB-231 cells. We detected five potential direct targets of miR-641 using qPCR (SRCAP, SIKE1, NADK, KHDC4, and HSPG2). SRCAP expression decreased significantly in miR-641 overexpression cells and the binding of SRCAP\'s 3\'UTR and miR-641 was further confirmed by dual-luciferase report gene assay. SRCAP blocked the proliferation and invasion inhibition induced by miR-641 or si-LINC00665 in MCF7 and MDA-MB-231 cells. In conclusion, LINC00665 could promote the survival and metastasis of breast cancer cells through sponging miR-641 and targeting SRCAP. This research provided new potential targets for targeted therapy in human breast cancer.