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CDCA7L 通过靶向 CCND1 促进神经胶质瘤增殖,并预测预后不良。

CDCA7L promotes glioma proliferation by targeting CCND1 and predicts an unfavorable prognosis.

机构信息

Department of Neurosurgery, The First Affiliated Hospital of Xinxiang Medical University, Xinxiang, Henan 453100, P.R. China.

Department of Emergency Medicine, The First Affiliated Hospital of Xinxiang Medical University, Xinxiang, Henan 453100, P.R. China.

出版信息

Mol Med Rep. 2019 Aug;20(2):1149-1156. doi: 10.3892/mmr.2019.10349. Epub 2019 Jun 5.

Abstract

Cell division cycle associated 7 like (CDCA7L) belongs to the JPO protein family, recently identified as a target gene of c‑Myc and is frequently dysregulated in multiple cancers. However, to the best of our knowledge, no studies to date have been carried out to investigate the functions of CDCA7L in glioma. Thus, in this study, the expression level of CDCA7L and its association with the prognosis in glioma were detected through the TCGA database. The mRNA expression levels of CDCA7L in glioblastoma (GBM) tissues and normal brain tissues were detected by RT‑qPCR and western blot analysis. To explore the role of CDCA7L in glioma, CDCA7L siRNA was constructed and transfected into U87 glioma cells. The expression levels of CDCA7L and cyclin D1 (CCND1) in glioma U87 cells following transfection with CDCA7L siRNA were measured by RT‑qPCR and western blot analysis. CCK‑8, colony formation, EdU and Transwell assays were used to measure the effects of CDCA7L on U87 cell proliferation, and flow cytometry was used to monitor the changes in the cell cycle following transfection with CDCA7L siRNA. Xenograft tumors were examined in vivo for the carcinogenic effects, as well as the mechanisms and prognostic value of CDCA7L in glioma tissues. The results revealed that CDCA7L was highly expressed in human GBM tissues, and a high expression of CDCA7L was associated with a poor prognosis of glioma patients through the TCGA database. We demonstrated that CDCA7L was highly expressed in human GBM tissues and 3 glioma cell lines. The downregulation CDCA7L expression significantly inhibited the proliferation and colony formation ability of U87 cells by blocking cell cycle progression in the G0/G1 phase. In addition, we found that the mRNA and protein levels of CCND1 were markedly decreased following transfection with CDCA7L siRNA compared with NC siRNA in vitro. The downregulation CDCA7L expression reduced the number of invading cells. Consistent with the results of the in vitro assays, the xenograft assay, immunohistochemistry (IHC) assay and western blot analysis demonstrated that, in response to CDCA7L inhibition, tumor growth was inhibited, Ki‑67 and CCND1 expression levels were decreased in vivo. On the whole, the results of the current study indicate that CDCA7L is highly expressed in human glioma tissues and that a high CDCA7L expression predicts a poor prognosis of glioma patients. CDCA7L promotes glioma U87 cell growth through CCND1.

摘要

细胞分裂周期相关蛋白 7 样(CDCA7L)属于 JPO 蛋白家族,最近被确定为 c-Myc 的靶基因,在多种癌症中经常失调。然而,据我们所知,目前尚无研究探讨 CDCA7L 在神经胶质瘤中的功能。因此,在本研究中,我们通过 TCGA 数据库检测了 CDCA7L 的表达水平及其与神经胶质瘤预后的关系。通过 RT-qPCR 和 Western blot 分析检测 CDCA7L 在脑胶质瘤(GBM)组织和正常脑组织中的 mRNA 表达水平。构建 CDCA7L siRNA 并转染 U87 神经胶质瘤细胞,探讨 CDCA7L 在神经胶质瘤中的作用。通过 RT-qPCR 和 Western blot 分析检测转染 CDCA7L siRNA 后神经胶质瘤 U87 细胞中 CDCA7L 和细胞周期蛋白 D1(CCND1)的表达水平。CCK-8、集落形成、EdU 和 Transwell 实验用于测量 CDCA7L 对 U87 细胞增殖的影响,流式细胞术用于监测转染 CDCA7L siRNA 后细胞周期的变化。体内检测异种移植肿瘤的致癌作用,以及 CDCA7L 在神经胶质瘤组织中的机制和预后价值。结果表明,CDCA7L 在人 GBM 组织中高表达,通过 TCGA 数据库发现 CDCA7L 高表达与神经胶质瘤患者预后不良相关。我们证明 CDCA7L 在人 GBM 组织和 3 种神经胶质瘤细胞系中高表达。下调 CDCA7L 表达通过阻滞 G0/G1 期细胞周期进展显著抑制 U87 细胞的增殖和集落形成能力。此外,我们发现与 NC siRNA 相比,转染 CDCA7L siRNA 后 U87 细胞中 CCND1 的 mRNA 和蛋白水平明显降低。下调 CDCA7L 表达可减少侵袭细胞的数量。与体外实验结果一致,异种移植实验、免疫组织化学(IHC)检测和 Western blot 分析表明,在体内抑制 CDCA7L 表达可抑制肿瘤生长,降低 Ki-67 和 CCND1 的表达水平。综上所述,本研究结果表明,CDCA7L 在人神经胶质瘤组织中高表达,CDCA7L 高表达预示着神经胶质瘤患者预后不良。CDCA7L 通过 CCND1 促进神经胶质瘤 U87 细胞生长。

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