Introduction: Long non-coding RNAs (LncRNAs) play a critical role in development and progression of various cancers. More and more researchers pay attention to the effect of lncRNA on regulating the cancer. However, the function and mechanism of Duxap8 in colorectal cancer have not been studied.
Methods: Reverse transcription quantitative PCR (RT-qPCR), cell counting kit-8 (CCK-8), 5-ethynyl-20-deoxyuridine (EdU), colony formation assay, flow cytometry, TdT-mediated dUTP nick-end labeling (TUNEL), Western blot, hematoxylin-eosin staining (HE), in situ hybridization (ISH) analysis, immunohistochemistry (IHC) and tumor transplantation experiment were performed to investigate the function and mechanism of Duxap8 in colorectal cancer.
Results: We found that the expression level of Duxap8 in colorectal cancer was closely correlated with tumor size (P = 0.024), tumor depth (P = 0.035) and lymphatic invasion (P =0.067) among 50 colorectal cancer patients. Then, we proved that the expression level of Duxap8 was significantly increased in human colorectal cancer tissues and cell lines. Functionally, Duxap8 knockdown inhibited the proliferation and induced the apoptosis of colorectal cancer cells, while Duxap8 overexpression facilitated the proliferation and suppressed the apoptosis in colorectal cancer in vitro. Moreover, knockdown of Duxap8 inhibited the size and weight of tumors in mice injected with colorectal cancer cells, overexpression of Duxap8 promoted the growth of colorectal cancer cells in vivo. Mechanically, we found that Duxap8 was principally located in the cytoplasm. Furthermore, Duxap8 functioned as a competing endogenous RNA to induce the development and progression of colorectal cancer through sponging miR-519b-3p to upregulate ZNF277.
Discussion: Taken together, our results demonstrated that Duxap8 enhanced the expression level of ZEB1 to promote via competing for miR-519b-3p, which might be a promising molecular therapeutic target of colorectal cancer.

Keywords: Duxap8, miR-519b-3p, ZNF277, colorectal cancer


中文翻译：

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长非编码 RNA Duxap8 通过 miR-519b/ZNF277 轴促进结直肠癌的细胞增殖并诱导细胞凋亡

简介：长链非编码 RNA (LncRNA) 在各种癌症的发展和进展中发挥着关键作用。越来越多的研究人员关注lncRNA对癌症的调控作用。然而，Duxap8在结直肠癌中的作用和机制尚未研究。
方法：逆转录定量PCR（RT-qPCR）、细胞计数试剂盒8（CCK-8）、5-乙炔基-20-脱氧尿苷（EdU）、集落形成试验、流式细胞术、TdT介导的dUTP缺口末端标记（采用TUNEL)、Western blot、苏木精-伊红染色(HE)、原位杂交(ISH)分析、免疫组化(IHC)和肿瘤移植实验研究Duxap8在结直肠癌中的作用和机制。
结果：在50例大肠癌患者中，我们发现Duxap8在大肠癌中的表达水平与肿瘤大小（P = 0.024）、肿瘤深度（P = 0.035）和淋巴浸润（P = 0.067）密切相关。然后，我们证明了Duxap8在人结直肠癌组织和细胞系中的表达水平显着增加。在功能上，Duxap8敲低抑制了结直肠癌细胞的增殖并诱导了其凋亡，而Duxap8过表达促进了结直肠癌细胞的增殖并抑制了体外凋亡。此外，Duxap8的击倒抑制注射结直肠癌细胞的小鼠中肿瘤的大小和重量，Duxap8的过表达促进了体内结直肠癌细胞的生长。机械地，我们发现Duxap8主要位于细胞质中。此外，Duxap8作为一种竞争性内源性 RNA，通过海绵状 miR-519b-3p 上调 ZNF277 来诱导结直肠癌的发展和进展。
讨论：综上所述，我们的结果表明，Duxap8通过竞争 miR-519b-3p 增强了 ZEB1 的表达水平，从而促进了结直肠癌的分子治疗靶点。

关键词： Duxap8, miR-519b-3p, ZNF277, 结直肠癌