Liu Zhen, Wang Xiaoyu, Liu Lin, Guan Miaosheng, Jiang Hao, An Dan, Li Hongbo
Department of Stomatology, the Eighth Medical Center of Chinese PLA General Hospital, No.17 heishanhu Road, Haidian District, Beijing 100091, China.
Department of Stomatology, PLA Strategic Support Force Medical Center, No.9 Anxiang Beili, Deshengmenwai, Chaoyang District, Beijing 100101, China.
Arch Oral Biol. 2023 Oct;154:105762. doi: 10.1016/j.archoralbio.2023.105762. Epub 2023 Jul 11.
This article aims to elucidate the role of Long non-coding RNA SLC7A11 antisense RNA1 (SLC7A11-AS1) in oral squamous cell carcinoma, which are expected to be useful for the oral squamous cell carcinoma diagnosis and treatment.
SLC7A11-AS1 expression was detected by quantitative real-time reverse transcription polymerase chain reaction (qRT-PCR) in oral squamous cell carcinoma cell lines. Cellular localization of SLC7A11-AS1C was detected by fluorescence in situ hybridization (FISH) assays and subcellular fractionation assay. Biological functions of SLC7A11-AS1 were explored by 3-(4,5-dimethyl-2-thiazolyl)- 2,5-diphenyl-2-H-tetrazolium bromide (MTT), 5-ethynyl-2'-deoxyuridine (EdU), wounding healing, and transwell invasion assays in vitro, as well as mice xenograft experiments and metastasis assays in vivo. RNA pull-down, RNA immunoprecipitation, co-immunoprecipitation, ubiquitination assays, and rescue experiments were performed to determine the molecular mechanism of SLC7A11-AS1 in oral squamous cell carcinoma.
SLC7A11-AS1 is overexpressed in oral cancer tissues and cell lines. Functionally, knockdown of SLC7A11-AS1 reduced the proliferation, migration, and invasion of oral squamous cell carcinoma cells in vitro and inhibited tumor growth as well as metastasis in vivo. Mechanistically, SLC7A11-AS1 impeded the interaction between K-homology type splicing regulatory protein (KHSRP) and kelch-like 12 (KLHL12), maintaining the stability of KHSRP by restraining KHSRP degradation through the ubiquitination-proteasome pathway. Furthermore, KHSRP overexpression recovered the malignant behaviors inhibited by SLC7A11-AS1 knockdown in oral cancer cells.
SLC7A11-AS1 promoted oral squamous cell carcinoma development by interacting with KHSRP and maintaining KHSRP stability by preventing its degradation via the ubiquitination-proteasome pathway. Thus, SLC7A11-AS1 is a potential therapeutic target for oral cancer.
本文旨在阐明长链非编码RNA SLC7A11反义RNA1（SLC7A11-AS1）在口腔鳞状细胞癌中的作用，有望对口腔鳞状细胞癌的诊断和治疗有所帮助。
采用定量实时逆转录聚合酶链反应（qRT-PCR）检测口腔鳞状细胞癌细胞系中SLC7A11-AS1的表达。通过荧光原位杂交（FISH）分析和亚细胞分级分离分析检测SLC7A11-AS1C的细胞定位。通过体外3-(4,5-二甲基-2-噻唑基)-2,5-二苯基-2-H-四氮唑溴盐（MTT）、5-乙炔基-2'-脱氧尿苷（EdU）、伤口愈合和transwell侵袭试验，以及体内小鼠异种移植实验和转移试验，探讨SLC7A11-AS1的生物学功能。进行RNA下拉、RNA免疫沉淀、共免疫沉淀、泛素化试验和挽救实验，以确定SLC7A11-AS1在口腔鳞状细胞癌中的分子机制。
SLC7A11-AS1在口腔癌组织和细胞系中过表达。在功能上，敲低SLC7A11-AS1可降低口腔鳞状细胞癌细胞在体外的增殖、迁移和侵袭能力，并在体内抑制肿瘤生长和转移。机制上，SLC7A11-AS1阻碍了K-同源性剪接调节蛋白（KHSRP）与kelch样蛋白12（KLHL12）之间的相互作用，通过抑制KHSRP通过泛素化-蛋白酶体途径的降解来维持KHSRP的稳定性。此外，KHSRP过表达恢复了口腔癌细胞中因SLC7A11-AS1敲低而受到抑制的恶性行为。
SLC7A11-AS1通过与KHSRP相互作用并通过泛素化-蛋白酶体途径防止其降解来维持KHSRP稳定性，从而促进口腔鳞状细胞癌的发展。因此，SLC7A11-AS1是口腔癌潜在的治疗靶点。
