State Key Laboratory of Oral Diseases, Department of Orthodontics, National Clinical Research Center for Oral Diseases, Other Research Platforms, West China Hospital of Stomatology, Sichuan University, Chengdu, China.
Department of Neurosurgery, West China Hospital, Sichuan University, Chengdu, China.
J Oral Pathol Med. 2019 Nov;48(10):919-928. doi: 10.1111/jop.12936. Epub 2019 Aug 18.
Malignant tumors arising from the epithelium of the oral cavity are termed as squamous cell carcinomas (OSCC). The aim of the current work was to understand the role of an isoform of RAS-like protein (RAL), RALBP1, in mediating squamous cell tumorigenesis. The study also aims to understand epigenetic modifications of RALBP1 mediated through microRNA-148a/b-3p.
Biopsies of tumor and healthy tissues from 25 patients with OSCC were collected and subjected to RNA and protein extraction to confirm upregulation of RLBP1 in tumor tissues. Expression of RLBP1 was silenced in SCC-9, using shRNA, and HN6 was transfected with plasmid bearing genes for RLBP1 over expression. Tumorigenic traits such as increased glucose uptake, aerobic glycolysis, enhanced cellular survival, cell migration, and invasion were assessed. Probable, molecular machinery involved in the upregulation was also assessed using Western blots. Using Target Scan tool, the miRNAs targeting RLBP1 were identified. Rescue of phenotypes in presence of miRNAs were also evaluated.
Over expression of RLBP1 was associated with increased glucose uptake and aerobic glycolysis mediated ATP synthesis. The cells divided at a faster rate with a higher rate of migration and invasion phenotype. miR-148a/b-3p were found to target RLBP1 and rescued RLBP1 mediated phenotype.
RLBP1 may mediate squamous cell tumorigenesis in oral cavity, independently of the RAS protein and through Akt. miR-148a/b-3p functions as a tumor suppressor by targeting RLBP1.
起源于口腔上皮的恶性肿瘤被称为鳞状细胞癌（OSCC）。本研究旨在探讨 Ras 样蛋白（RAL）同工型 RALBP1 在介导鳞状细胞肿瘤发生中的作用，并了解通过 microRNA-148a/b-3p 介导的 RALBP1 的表观遗传修饰。
收集 25 例 OSCC 患者的肿瘤和健康组织活检标本，提取 RNA 和蛋白质，以证实 RLBP1 在肿瘤组织中的上调。使用 shRNA 沉默 SCC-9 中的 RLBP1 表达，并用携带 RLBP1 过表达基因的质粒转染 HN6。评估葡萄糖摄取增加、有氧糖酵解、增强细胞存活、细胞迁移和侵袭等肿瘤发生特性。还使用 Western blot 评估可能涉及的分子机制。使用 TargetScan 工具鉴定靶向 RLBP1 的 miRNAs。还评估了存在 miRNAs 时表型的恢复情况。
RLBP1 的过表达与增加的葡萄糖摄取和有氧糖酵解介导的 ATP 合成有关。细胞以更快的速度分裂，具有更高的迁移和侵袭表型。miR-148a/b-3p 被发现靶向 RLBP1，并挽救了 RLBP1 介导的表型。
RLBP1 可能通过 Akt 独立于 Ras 蛋白介导口腔鳞状细胞肿瘤发生。miR-148a/b-3p 通过靶向 RLBP1 发挥肿瘤抑制作用。
