目的：检测lncRNA DNM3OS在喉鳞状细胞癌（laryngeal squamous cell carcinoma，LSCC）组织和LSCC细胞株中的表达及其临床意义，探讨其对LSCC TU177细胞体外增殖、迁移及侵袭的影响，并分析DNM3OS与EMT的关系。方法：从河北医科大学第四医院生物标本库选取2014年3月至2018年12月收治的68例LSCC患者手术切除的癌及癌旁组织标本，应用qPCR法检测DNM3OS在LSCC组织和细胞株中的表达水平。采用siRNA敲低TU177细胞中DNM3OS的表达，应用MTS、克隆形成及Transwell小室等方法分别检测敲低DNM3OS表达对TU177细胞增殖、迁移和侵袭等生物学行为的影响。应用qPCR和WB法检测转染si-DNM3OS后对EMT标志物上皮钙黏素（E-cadherin）、神经钙黏素（N-cadherin）、波形蛋白（vimentin）、扭曲蛋白（twist）、锌指转录因子2（SNAI2）mRNA和蛋白的变化。结果：LSCC组织中DNM3OS表达水平明显高于癌旁组织（P<0.01），并与患者的TNM分期、淋巴结转移及生存期有关联（P<0.05 或 P<0.01）。DNM3OS在LSCC细胞株（Hep-2、AMC-HN-8、TU177、TU212及TU686）中均呈现不同程度的高表达（P<0.05 或 P<0.01），转染si-DNM3OS后TU177细胞中DNM3OS的表达显著降低（P<0.01）。与对照组相比，DNM3OS表达敲低可抑制TU177细胞的体外增殖、迁移和侵袭能力（P<0.05 或 P<0.01），可上调 TU177 细胞中E-cadherin的表达而下调 N-cadherin、vimentin、twist和SNAI2的表达（均P<0.01）。结论：DNM3OS高表达与LSCC的恶性进展有关，其可能为预测LSCC患者预后的潜在指标；DNM3OS可能通过影响EMT进程促进LSCC细胞的侵袭和转移。

Objective: To explore the expression and clinical significance of lncRNA DNM3OS (dynamin 3 opposite strand/antisense RNA) in laryngeal squamous cell carcinoma (LSCC) tissues and cell lines, and to investigate its effects on in vitro proliferation,migration and invasion of LSCC TU177 cells, as well as to discuss the relationship between DNM3OS and epithelial-mesenchymal transition (EMT). Methods: Sixty-eight pairs of cancer and corresponding para-cancerous tissues from LSCC patients that admitted for surgery from March 2014 to December 2018 were collected from the biological specimen bank of the Fourth hospital of Hebei Medical University. The level of DNM3OS expression in LSCC tissues and cell lines was detected by qPCR. siRNA was used to knockdown DNM3OS expression in TU177 cell. MTS, colony formation and Transwell chamber assays were performed to detect the effect of DNM3OS knockdown on proliferation, migration and invasion of TU177 cells. qPCR and WB methods were used to detect the mRNA and protein expression of EMT-related markers, such as E-cadherin, N-cadherin, vimentin, twist, and SNAI2 after DNM3OS knockdown. Results: The expression level of DNM3OS in LSCC tissues was markedly higher than that in para-cancerous tissues (P<0.01). Upregulated DNM3OS expression was related to TNM stage, lymph node metastasis, and survival of patients with LSCC (P<0.05 or P<0.01). Furthermore, DNM3OS was highly expressed in five LSCC cell lines (Hep-2, AMC-HN-8, TU177, TU212, and TU686) (P<0.05 or P<0.01). The expression of DNM3OS was significantly decreased in TU177 cells after transfection of si-DNM3OS (P<0.01). Compared with the control group, DNM3OS knockdown could suppress the in vitro proliferation, migration and invasion of TU177 cells (all P<0.01), upregulate the expression level of E-cadherin, while down-regulate the expression of N-cadherin,vimentin, twist and SNAI2 in TU177 cells (all P<0.01). Conclusion: DNM3OS overexpression is related to the malignant progression of LSCC, which may be a potential prognostic marker for LSCC patients. DNM3OS may promote the invasion and metastasis of LSCC cells by mediating EMT.

河北省医学科学研究重点课题资助项目（No. 20180588, No. 20201511）