Wang Yizhi, Yuan Da, Zhou Li, Liang Zhiyong, Zhou Weixun, Lu Jun, Jiang Bolun, You Lei, Guo Junchao, Zhao Yu-Pei
Department of General Surgery, Peking Union Medical College Hospital, Chinese Academy of Medical Sciences, Peking Union Medical College, Beijing, China.
Medical Management Office, Department of General Surgery, Peking Union Medical College Hospital, Chinese Academy of Medical Sciences, Peking Union Medical College, Beijing, China.
Front Oncol. 2020 May 5;10:576. doi: 10.3389/fonc.2020.00576. eCollection 2020.
Transducin-like enhancer of split-1 (TLE1), a member of the Groucho/TLE family of transcriptional corepressors, has been reported to be involved in the tumorigenesis of various cancers and function as a clinical prognostic indicator. However, the mechanisms and prognostic significance of TLE1 in pancreatic ductal adenocarcinoma (PDAC) have not been elucidated. In this study, western blot analyses and real-time polymerase chain reaction (RT-PCR) were employed to evaluate the expression of TLE1 and related proteins in PDAC cell lines. Wound healing, transwell migration and invasion, and Cell Counting Kit-8 (CCK-8) assays were used to determine cell line-specific differences in metastasis and proliferation. Flow cytometry was performed for cell cycle detection. RNA sequencing and bioinformatics were undertaken to explore the molecular mechanisms and potential targeted molecules of TLE1. TLE1 expression in tumor and para-tumor tissues was evaluated by tissue microarray-based immunohistochemistry using a semiquantitative method (-score) in 262 patients with radical PDAC resection. Correlation, Kaplan-Meier survival, univariate, and multivariate analyses were also performed. Our findings showed that TLE1 expression was common in PDAC cell lines. Upregulation of TLE1 inhibited PDAC cell migration, invasion, and proliferation by delaying the G0/G1 transition. Immunohistochemistry revealed that TLE1 was specifically expressed in the nucleus and at higher levels in tumor tissues compared with para-tumor tissues. Generally, high TLE1 expression was associated with no vascular invasion. In univariate analyses, high TLE1 expression was associated with longer disease-specific survival (DSS) in all patients and in 16 patient subgroups. In multivariate analyses, TLE1 expression was independently associated with DSS in all patients and four patient subgroups. In conclusion, these results suggest that TLE1 has an inhibitory role in PDAC progression and is a favorable prognostic indicator for patients with resectable PDAC.
分裂样转录增强子1（TLE1）是转录共抑制因子Groucho/TLE家族的成员，据报道它参与多种癌症的肿瘤发生，并作为临床预后指标发挥作用。然而，TLE1在胰腺导管腺癌（PDAC）中的作用机制和预后意义尚未阐明。在本研究中，采用蛋白质免疫印迹分析和实时聚合酶链反应（RT-PCR）来评估TLE1及相关蛋白在PDAC细胞系中的表达。使用伤口愈合实验、Transwell迁移和侵袭实验以及细胞计数试剂盒8（CCK-8）实验来确定细胞系在转移和增殖方面的特异性差异。通过流式细胞术进行细胞周期检测。进行RNA测序和生物信息学分析以探索TLE1的分子机制和潜在的靶向分子。使用基于组织芯片的免疫组织化学方法，采用半定量方法（-评分）评估262例接受根治性PDAC切除术患者的肿瘤组织和癌旁组织中TLE1的表达。还进行了相关性分析、Kaplan-Meier生存分析、单因素分析和多因素分析。我们的研究结果表明，TLE1在PDAC细胞系中普遍表达。TLE1的上调通过延迟G0/G1期转换来抑制PDAC细胞的迁移、侵袭和增殖。免疫组织化学显示，TLE1特异性表达于细胞核，与癌旁组织相比，在肿瘤组织中的表达水平更高。一般来说，TLE1高表达与无血管侵犯相关。在单因素分析中，TLE1高表达与所有患者以及16个患者亚组的疾病特异性生存期（DSS）延长相关。在多因素分析中，TLE1表达与所有患者以及4个患者亚组的DSS独立相关。总之，这些结果表明TLE1在PDAC进展中具有抑制作用，并且是可切除性PDAC患者的良好预后指标。
