Zhan Wei, Li Yuzhe, Liu Xuhui, Zheng Changlong, Fu Yongmei
Department of Emergency, The Third Affiliated Hospital of Sun Yat-Sen University, Guangzhou 510000, People's Republic of China.
Department of Clinical Laboratory, The Third Affiliated Hospital of Sun Yat-Sen University, Lingnan Hospital, Guangzhou 510080, People's Republic of China.
Cancer Manag Res. 2020 Jan 24;12:599-610. doi: 10.2147/CMAR.S235933. eCollection 2020.
Lung cancer is the most common cancer in the world and is the main cause of cancer-related death. Revealing the potential mechanism of malignant characteristics of lung cancer is urgent for treating this disease effectively. Zinc finger protein 671 (ZNF671) is a member of the largest transcription factor family in the human genome. The role of ZNF671 in non-small-cell lung cancer (NSCLC) remains unknown. The purpose of this study was to investigate the function and mechanism of ZNF671 in NSCLC.
ZNF671 expression in NSCLC cells and tissues were detected by Real-Time PCR, Western blot and TCGA databases. Then, we evaluated the prognostic value of ZNF671 expression in NSCLC using the Kaplan-Meier plotter (KM plotter) and TCGA databases. Moreover, the function of ZNF671 in the proliferation and metastasis of lung cancer was investigated by MTT assay, colony formation assay, in vivo experiment, EdU assay, wound healing assay, transwell assay, and 3D culture assay. Luciferase reporter and subcellular fractionation assays were performed to determine the underlying mechanism of ZNF671-mediated proliferation and metastasis of NSCLC.
ZNF671 expression was significantly reduced in both NSCLC cell lines and clinical specimens compared to that in normal controls. The survival analysis results indicated that the downregulation of ZNF671 significantly correlates with poor prognosis and predicts a shorter overall survival and post-progression survival among NSCLC patients. Ectopic overexpression of ZNF671 dramatically restrains, whereas silencing ZNF671 enhanced, cell proliferation and metastasis of NSCLC. Mechanically, gene set enrichment analysis (GSEA) showed that the expression of ZNF671 was significantly correlated with Wnt/β-catenin signaling. Simultaneously, our results confirm that the overexpression of ZNF671 inhibits cell cycle progression and metastasis by weakening the Wnt/β-catenin pathway, and then downregulating the expression of downstream target genes CyclinD1 and MMP9.
This study found that the overexpression of ZNF671 restrains the proliferation and metastasis of lung cancer through inhibiting Wnt/β-catenin signaling pathway. Furthermore, our current results provide important insights into ZNF671 as an excellent predictive biomarker for NSCLC, thus providing a novel perspective for the treatment of NSCLC.
肺癌是全球最常见的癌症，也是癌症相关死亡的主要原因。揭示肺癌恶性特征的潜在机制对于有效治疗该疾病至关重要。锌指蛋白671（ZNF671）是人类基因组中最大的转录因子家族的成员。ZNF671在非小细胞肺癌（NSCLC）中的作用尚不清楚。本研究的目的是探讨ZNF671在NSCLC中的功能和机制。
通过实时荧光定量PCR、蛋白质免疫印迹法和TCGA数据库检测NSCLC细胞和组织中ZNF671的表达。然后，我们使用Kaplan-Meier绘图仪（KM绘图仪）和TCGA数据库评估ZNF671表达在NSCLC中的预后价值。此外，通过MTT法、集落形成试验、体内实验、EdU试验、伤口愈合试验、Transwell试验和三维培养试验研究ZNF671在肺癌增殖和转移中的作用。进行荧光素酶报告基因和亚细胞分级分离试验以确定ZNF671介导的NSCLC增殖和转移的潜在机制。
与正常对照相比，NSCLC细胞系和临床标本中ZNF671的表达均显著降低。生存分析结果表明，ZNF671的下调与不良预后显著相关，并预测NSCLC患者的总生存期和进展后生存期较短。ZNF671的异位过表达显著抑制，而沉默ZNF671则增强NSCLC的细胞增殖和转移。机制上，基因集富集分析（GSEA）表明ZNF671的表达与Wnt/β-连环蛋白信号通路显著相关。同时，我们的结果证实ZNF671的过表达通过削弱Wnt/β-连环蛋白途径抑制细胞周期进程和转移，进而下调下游靶基因CyclinD1和MMP9的表达。
本研究发现ZNF671的过表达通过抑制Wnt/β-连环蛋白信号通路抑制肺癌的增殖和转移。此外，我们目前的结果为ZNF671作为NSCLC的优秀预测生物标志物提供了重要见解，从而为NSCLC的治疗提供了新的视角。
