机构：[1]Department of Otolaryngology–Head and Neck Surgery, Beijing Tongren Hospital, Capital Medical University, Beijing, China临床科室耳鼻咽喉-头颈外科首都医科大学附属北京同仁医院首都医科大学附属同仁医院[2]Department of Otolaryngology–Head and Neck Surgery, Affiliated Yantai Yuhuangding Hospital of Qingdao University, Yantai, Shandong Province, China
Objective: To evaluate the function of long non-coding RNA ANRIL (CDKN2B-AS1) in laryngeal squamous cell cancer (LSCC), and to explore the underlying mechanism. Methods: The expression levels of CDKN2B-AS1 in LSCC tissues and cell lines (Tu177, HN4, AMC-HN-8 and NP69) were determined by reverse transcription quantitative PCR (RT-qPCR). AMC-HN-8 cells were then transfected with siRNAs of CDKN2B-AS1. The effects of CDKN2B-AS1 on cell proliferation, cell cycle, and apoptotic protein were determined by CCK-8 assay, flow cytometry analysis, and western blot, respectively. Dual luciferase reporter assay and RNA immunoprecipitation (RIP) assay were employed to verify the targets of CDKN2B-AS1. The miR-324-5p mimics or miR-324-5p inhibitor and ROCK1 over-expression plasmids were also transfected into AMC-HN-8 cells for further analysis. Results: CDKN2B-AS1 was upregulated in LSCC tissues, and the upregulation of CDKN2B-AS1 was correlated with overall survival, advanced clinical stage, and lymph node metastasis. In AMC-HN-8 cells, the knockdown of CDKN2B-AS1 by siRNA inhibited cell viability, blocked cell cycle in G1 phase, and increased the expression levels of cyclin-dependent kinase inhibitor 1A (p21), cleaved caspase3, and cleaved PPoly (ADP-Ribose) polymerase 1. Results of dual luciferase reporter assay showed that miR-324-5p could bind to CDKN2B-AS1 or Rho-associated coiled-coil containing protein kinase 1 (ROCK1). Finally, over-expression of ROCK1 in AMC-HN-8 cells revised the inhibitory effect of CDKN2B-AS1 siRNA on cell growth. Discussion: The upregulation of CDKN2B-AS1 was correlated with overall survival, advanced clinical stage, and lymph node metastasis and promoted LSCC cell growth via miR-324-5p/ROCK1 axis.
基金：
National Key Research and Development Program of China [2017YFC0112500]; Beijing Natural Science FoundationBeijing Natural Science Foundation [7182036]; Special Fund of the Pediatric Medical Coordinated Development Center of Beijing Hospitals Authority [XTCX201823]
第一作者机构：[1]Department of Otolaryngology–Head and Neck Surgery, Beijing Tongren Hospital, Capital Medical University, Beijing, China[2]Department of Otolaryngology–Head and Neck Surgery, Affiliated Yantai Yuhuangding Hospital of Qingdao University, Yantai, Shandong Province, China
通讯作者：
通讯机构：[1]Department of Otolaryngology–Head and Neck Surgery, Beijing Tongren Hospital, Capital Medical University, Beijing, China[*1]Department of Otolaryngology–Head and Neck Surgery, Beijing Tongren Hospital, Capital Medical University, No. 1 Dongjiaominxiang, Beijing, 100730, China.
推荐引用方式(GB/T 7714)：
Liu Feifei,Xiao Yang,Ma Lijing,et al.Regulating of cell cycle progression by the lncRNA CDKN2B-AS1/miR-324-5p/ROCK1 axis in laryngeal squamous cell cancer[J].INTERNATIONAL JOURNAL OF BIOLOGICAL MARKERS.2020,35(1):47-56.doi:10.1177/1724600819898489.
APA：
Liu, Feifei,Xiao, Yang,Ma, Lijing&Wang, Jun.(2020).Regulating of cell cycle progression by the lncRNA CDKN2B-AS1/miR-324-5p/ROCK1 axis in laryngeal squamous cell cancer.INTERNATIONAL JOURNAL OF BIOLOGICAL MARKERS,35,(1)
MLA：
Liu, Feifei,et al."Regulating of cell cycle progression by the lncRNA CDKN2B-AS1/miR-324-5p/ROCK1 axis in laryngeal squamous cell cancer".INTERNATIONAL JOURNAL OF BIOLOGICAL MARKERS 35..1(2020):47-56
