FOXD2-AS1与喉鳞状细胞癌临床病理参数的关系及其对喉癌细胞增殖的作用

FOXD2-AS1is corelated with clinicopathological parameter of laryngeal carcinoma and promote cancer cell proliferation

目的:探究长链非编码RNA FOXD2-AS1在喉癌组织中的表达及其临床意义,并在喉癌细胞中探究其对细胞增殖的作用。方法:利用实时荧光定量PCR检测FOXD2-AS1在85例喉癌组织中的表达,利用单因素方差分析判断其表达与喉癌临床病理参数的关系,利用生物信息学技术探究FOXD2-AS1在头颈部肿瘤中的预后意义。利用siRNA技术干扰FOXD2-AS1在喉癌细胞TU686中的表达,并用MTT、克隆形成实验等探究FOXD2-AS1的生物学作用。使用双荧光素酶实验探究与FOXD2-AS1结合的microRNA。结果:长链非编码RNA FOXD2-AS1在喉癌组织中的表达显著高于正常组织(t=10.012,P<0.05),FOXD2-AS1的表达与喉癌侵犯的范围(T分期)显著相关(χ~2=6.41,P=0.016),与患者的年龄、性别、吸烟史、肿瘤原发部位、淋巴结转移(N分期)等无关。利用GEPIA对TCGA数据库中头颈部肿瘤进行生存分析显示,FOXD2-AS1高表达的患者预后较差(P=0.048)。在TU686细胞中使用siRNA干扰FOXD2-AS1的表达可下调细胞的克隆形成能力(t=8.053,P<0.05),MTT实验证实干扰FOXD2-AS1可下调喉癌细胞的增殖活性(t=9.337,P<0.05)。双荧光素酶实验显示FOXD2-AS1能与miR-206结合,MTT实验证实miR-206inhibitor的转染可显著促进喉癌细胞的增殖(t=4.200,P<0.05);与si-FOXD2-AS1相比,si-FOXD2-AS1+miR-206inhibitor组的喉癌细胞增殖显著增强(t=6.803,P<0.05)。结论:长链非编码RNA FOXD2-AS1与喉鳞状细胞癌的临床病理参数显著相关,其可通过结合miR-206促进喉癌细胞增殖。

Objective: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.Method: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.Result:The expression of FOXD2-AS1 in laryngeal cancer tissues was significantly higher than that in normal tissues(t=10.012,P<0.05),and was associated with T staging of laryngeal cancer(χ~2=6.41,P=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(P=0.048).Suppressing FOXD2-AS1 via siRNA in TU686 cells decreased clonal formation ability(t=8.053,P<0.05)and MTT assay confirmed that interference of FOXD2-AS1 down-regulated proliferation activity of TU686 cells(t=9.337,P<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.Conclusion:FOXD2-AS1 is corelated with clinicopathological parameter of laryngeal squamous cell carcinoma and promotes cancer cell proliferation through targeting miR-206.