Chen W, Sun S G, Jiang M X, Li S S, Yuan K
Department of Otolaryngology Head and Neck Surgery, the Central Hospital of Wuhan, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430014, China.
Lin Chuang Er Bi Yan Hou Tou Jing Wai Ke Za Zhi. 2019 May;33(5):436-440. doi: 10.13201/j.issn.1001-1781.2019.05.013.
To investigate the expression and clinical significance of long non-coding RNA FOXD2-AS1 in laryngeal squamous cell carcinoma and its effect on cancer cell proliferation. Real-time quantitative polymerase chain reaction(RT-qPCR) was used to detect the expression of FOXD2-AS1 in 85 cases of laryngeal carcinoma. One-way ANOVA was used to determine relationships between its expression and clinicopathological parameters of laryngeal carcinoma. The prognostic significance of FOXD2-AS1 in head and neck cancer was explored using bioinformatics technology. SiRNA was used to interfere the expression of FOXD2-AS1 in TU686 laryngeal carcinoma cells. MTT and clonal formation assay were used to investigate the biological effect of FOXD2-AS1. MicroRNA binding to FOXD2-AS1 was investigated using double luciferase assay. The expression of FOXD2-AS1 in laryngeal cancer tissues was significantly higher than that in normal tissues(=10.012, <0.05), and was associated with T staging of laryngeal cancer(=6.41, =0.016). There were no relationships between FOXD2-AS1 expression and age, sex, smoking history, primary site of tumor and lymph node metastasis(N staging). Survival analysis of head and neck tumors in the TCGA database using GEPIA showed poor prognosis in patients with high FOXD2-AS1 expression(=0.048). Suppressing FOXD2-AS1 via siRNA in TU686 cells decreased clonal formation ability(=8.053, <0.05) and MTT assay confirmed that interference of FOXD2-AS1 down regulated proliferation activity of TU686 cells(=9.337, <0.05). Double luciferase assay showed that FOXD2-AS1 could directly bind to miR-206, thus inhibiting the expression of miR-206. Further MTT assay indicated that inhibiting miR-206 attenuated the suppressing effect of si-FOXD2-AS1 on the proliferation of TU686 cells. FOXD2-AS1 is corelated with clinicopathological parameter of laryngeal squamous cell carcinoma and promotes cancer cell proliferation through targeting miR-206.
探讨长链非编码RNA FOXD2-AS1在喉鳞状细胞癌中的表达及其临床意义，以及其对癌细胞增殖的影响。采用实时定量聚合酶链反应（RT-qPCR）检测85例喉癌组织中FOXD2-AS1的表达。采用单因素方差分析确定其表达与喉癌临床病理参数之间的关系。利用生物信息学技术探讨FOXD2-AS1在头颈癌中的预后意义。使用小干扰RNA（SiRNA）干扰TU686喉癌细胞中FOXD2-AS1的表达。采用MTT法和克隆形成实验研究FOXD2-AS1的生物学效应。利用双荧光素酶报告基因实验研究与FOXD2-AS1结合的微小RNA。喉癌组织中FOXD2-AS1的表达明显高于正常组织（F=10.012，P<0.05），且与喉癌的T分期相关（F=6.41，P=0.016）。FOXD2-AS1的表达与年龄、性别、吸烟史、肿瘤原发部位及淋巴结转移（N分期）无关。使用GEPIA对TCGA数据库中的头颈肿瘤进行生存分析，结果显示FOXD2-AS1高表达患者的预后较差（P=0.048）。通过SiRNA抑制TU686细胞中FOXD2-AS1的表达可降低其克隆形成能力（F=8.053，P<0.05），MTT实验证实干扰FOXD2-AS1可下调TU686细胞的增殖活性（F=9.337，P<0.05）。双荧光素酶报告基因实验表明，FOXD2-AS1可直接与miR-206结合，从而抑制miR-206的表达。进一步的MTT实验表明，抑制miR-206可减弱si-FOXD2-AS1对TU686细胞增殖的抑制作用。FOXD2-AS1与喉鳞状细胞癌的临床病理参数相关，并通过靶向miR-206促进癌细胞增殖。
