Wang Qiuling, Han Jingxin, Xu Pu, Jian Xinchun, Huang Xieshan, Liu Deyu
Stomatology Center, Affiliated Haikou Hospital of Xiangya Medical College, Central South University, Haikou, Hainan, 570208, People's Republic of China.
Department of Oral and Maxillofacial Surgery, Xiangya Hospital, Central South University, Central South University, Changsha, Hunan, 410008, People's Republic of China.
Cancer Manag Res. 2021 Feb 22;13:1831-1841. doi: 10.2147/CMAR.S298236. eCollection 2021.
Targeting the long non-coding RNAs (LncRNAs)-microRNAs (miRNAs)-mRNA competing endogenous RNA (ceRNA) networks has been proved as an effective strategy to treat multiple cancers, including oral squamous cell carcinoma (OSCC). Based on this, the present study identified a novel LncRNA SNHG16/miR-17-5p/CCND1 signaling pathway that played an important role in regulating the pathogenesis of OSCC.
The expression levels of cancer-associated genes were examined by Real-Time qPCR and Western Blot at transcriptional and translated levels, respectively. CCK-8 assay was performed to determine cell proliferation, and cell apoptosis ratio was measured by the Annexin V-FITC/PI double staining assay. Transwell assay was performed to examine cell migration, and dual-luciferase reporter gene system assay was used to validate the ceRNA networks.
LncRNA SNHG16 and CCND1 were upregulated, while miR-17-5p was downregulated in OSCC tissues and cell lines, compared to their normal counterparts. Also, miR-17-5p negatively correlated with both LncRNA SNHG16 and CCND1 mRNA, but LncRNA SNHG16 was positively relevant to CCND1 mRNA in OSCC tissues. By performing the gain- and loss-of-function experiments, we noticed that LncRNA SNHG16 overexpression aggravated the malignant phenotypes, such as cell proliferation, viability, migration and epithelial-mesenchymal transition (EMT) in OSCC cells, while LncRNA SNHG16 knock-down had opposite effects. Furthermore, our dual-luciferase reporter gene system evidenced that LncRNA SNHG16 sponged miR-17-5p to upregulate CCND1 in OSCC cells, and the inhibiting effects of LncRNA SNHG16 ablation on OSCC progression were abrogated by both downregulating miR-17-5p and overexpressing CCND1. Finally, the xenograft tumor-bearing mice models were established, and our data validated that LncRNA SNHG16 served as an oncogene to promote tumorigenicity of OSCC cells in vivo.
Taken together, targeting the LncRNA SNHG16/miR-17-5p/CCND1 axis hindered the development of OSCC, and this study provided potential diagnostic and therapeutic biomarkers for OSCC in clinic.
靶向长链非编码RNA（LncRNAs）-微小RNA（miRNAs）-信使核糖核酸（mRNA）竞争性内源RNA（ceRNA）网络已被证明是治疗包括口腔鳞状细胞癌（OSCC）在内的多种癌症的有效策略。基于此，本研究确定了一种新型的LncRNA SNHG16/miR-17-5p/CCND1信号通路，该通路在调节OSCC发病机制中发挥重要作用。
分别通过实时定量聚合酶链反应（Real-Time qPCR）和蛋白质免疫印迹法（Western Blot）在转录水平和翻译水平检测癌症相关基因的表达水平。采用细胞计数试剂盒-8（CCK-8）法测定细胞增殖情况，通过膜联蛋白V-异硫氰酸荧光素/碘化丙啶（Annexin V-FITC/PI）双染法检测细胞凋亡率。采用Transwell法检测细胞迁移情况，并使用双荧光素酶报告基因系统实验验证ceRNA网络。
与正常组织和细胞系相比，LncRNA SNHG16和细胞周期蛋白D1（CCND1）在OSCC组织和细胞系中表达上调，而miR-17-5p表达下调。此外，在OSCC组织中，miR-17-5p与LncRNA SNHG16和CCND1 mRNA均呈负相关，但LncRNA SNHG16与CCND1 mRNA呈正相关。通过进行功能获得和功能缺失实验，我们发现LncRNA SNHG16过表达加剧了OSCC细胞的恶性表型，如细胞增殖、活力、迁移和上皮-间质转化（EMT），而敲低LncRNA SNHG16则产生相反的效果。此外，我们的双荧光素酶报告基因系统证明，在OSCC细胞中，LncRNA SNHG16通过吸附miR-17-5p上调CCND1，下调miR-17-5p和过表达CCND1均可消除LncRNA SNHG16缺失对OSCC进展的抑制作用。最后，建立了荷瘤小鼠模型，我们的数据验证了LncRNA SNHG16作为一种癌基因在体内促进OSCC细胞的致瘤性。
综上所述，靶向LncRNA SNHG16/miR-17-5p/CCND1轴可阻碍OSCC的发展，本研究为临床OSCC提供了潜在的诊断和治疗生物标志物。
