Xu Bing, Chen Hui, Sun Xinchen, Cheng Hongyan, Chipusu Kavimbi, Hua Wei, Chen Tingting
Department of Oncology, Northern Jiangsu People's Hospital, Yangzhou, China.
Department of Radiation Oncology, The First Affiliated Hospital of Nanjing Medical University, Nanjing, China.
Front Bioeng Biotechnol. 2025 Jul 25;13:1604859. doi: 10.3389/fbioe.2025.1604859. eCollection 2025.
Esophageal cancer (EC) is one of the most common cancers worldwide. The prognosis for patients with the same stage of EC can vary substantially. Recurrence and metastasis after treatment are still important reasons for poor prognosis of esophageal cancer patients. Rhomboid domain containing 1 (RHBDD1) has been reported to play an important role in the development and progression of various cancers, but its role in esophageal malignancy is poorly understood, and this paper aims to explore the role of RHBDD in esophageal squamous cell carcinoma.
This study employed and approaches to investigate molecular mechanisms in ESCC. ECA109 cells were cultured in RPMI 1640 with 10% FBS under 5% CO at 37°C. RNA extraction (Trizol) and qRT-PCR (SYBR Green, β-actin normalization) were performed in triplicate. Lentiviral shRNA constructs targeting RHBDD1/ELK3 (GenePharma) were transfected, with stable clones selected via puromycin and validated by Western blot/qRT-PCR. Proliferation was assessed via CCK-8 (absorbance at 450 nm) and EdU assays (Ribo-Bio kit), while apoptosis was quantified by annexin V-FITC/PI staining using flow cytometry. Immunofluorescence detected β-catenin localization (Abcam antibodies). For analysis, BALB/c nude mice (n = 6/group) received subcutaneous ESCC xenografts, monitored biweekly for tumor volume (L × W/2). IHC evaluated protein expression (Ki67, EMT markers). Data, presented as mean ± SD, were analyzed by Student's t-test or ANOVA (Dunnett's ; p < 0.05). Protocols followed institutional ethical guidelines.
RHBDD1 promotes cell invasion and migration in ESCC cells. Furthermore, knockdown of RHBDD1 in ESCC cells reduced lung and liver metastasis . The results also indicated that RHBDD1 could promote cell proliferation and inhibit cell apoptosis, which may make ESCC cells more aggressive.
The present study shows that RHBDD1 is an activator of epithelial-mesenchymal transition. This study contributes to the understanding of the role of RHBDD1 in ESCC patients and serves as a valuable resource for in-depth exploration of the pathogenesis of ESCC and the identification of potential therapeutic targets in the future.
食管癌（EC）是全球最常见的癌症之一。相同分期的食管癌患者预后差异很大。治疗后的复发和转移仍是食管癌患者预后不良的重要原因。据报道，含菱形结构域1（RHBDD1）在多种癌症的发生和发展中起重要作用，但其在食管恶性肿瘤中的作用尚不清楚，本文旨在探讨RHBDD在食管鳞状细胞癌中的作用。
本研究采用 和 方法研究食管鳞状细胞癌的分子机制。ECA109细胞在含10%胎牛血清的RPMI 1640培养基中，于37°C、5%二氧化碳条件下培养。RNA提取（Trizol法）和qRT-PCR（SYBR Green法，以β-肌动蛋白标准化）重复进行3次。转染靶向RHBDD1/ELK3的慢病毒shRNA构建体（GenePharma公司），通过嘌呤霉素筛选稳定克隆，并通过蛋白质免疫印迹/qRT-PCR验证。通过CCK-8法（450nm处吸光度）和EdU检测法（Ribo-Bio试剂盒）评估细胞增殖，同时使用流式细胞术通过膜联蛋白V-FITC/PI染色定量细胞凋亡。免疫荧光检测β-连环蛋白的定位（Abcam抗体）。对于 分析，BALB/c裸鼠（每组n = 6）皮下接种食管鳞状细胞癌异种移植物，每两周监测一次肿瘤体积（长×宽/2）。免疫组化评估蛋白表达（Ki67、上皮-间质转化标志物）。数据以平均值±标准差表示，采用学生t检验或方差分析（Dunnett检验；p < 0.05）。实验方案遵循机构伦理准则。
RHBDD1促进食管鳞状细胞癌细胞的侵袭和迁移。此外，敲低食管鳞状细胞癌细胞中的RHBDD1可减少肺和肝转移。结果还表明，RHBDD1可促进细胞增殖并抑制细胞凋亡，这可能使食管鳞状细胞癌细胞更具侵袭性。
本研究表明RHBDD1是上皮-间质转化的激活剂。本研究有助于理解RHBDD1在食管鳞状细胞癌患者中的作用，并为未来深入探索食管鳞状细胞癌的发病机制和确定潜在治疗靶点提供有价值的资源。
