LncRNA TUG1通过调控miR-196b-5p调控卵巢癌细胞凋亡和放射敏感性分子作用机制

目的 探讨LncRNA TUG1通过调控miR-196b-5p表达对卵巢癌细胞凋亡和放射敏感性的影响及其潜在的作用机制。方法 运用qRT-PCR和Western blot检测卵巢癌细胞株SKOV3、HO8910、OVCAR3和正常卵巢上皮细胞HOSE中miR-196b-5p和TUG1的表达水平。双荧光素酶报告基因分析法验证TUG1可能的靶基因。建立TUG1抑制表达、miR-196b-5p过表达及si-TUG1和anti-miR-196b-5p共转染的SKOV3细胞株,流式细胞术检测细胞凋亡率。采用克隆形成实验检测各组SKOV3细胞不同剂量X射线照射后的存活情况,曲线拟合计算不同剂量辐射后各放射生物学参数。结果 正常卵巢上皮细胞HOSE相比,卵巢癌细胞SKOV3、HO8910及OVCAR3中miR-196b-5p的表达显著降低,差异有统计学意义(P<0.05),TUG1的表达显著升高,差异有统计学意义(P<0.05)。miR-196b-5p是TUG1的靶基因。TUG1抑制表达或miR-196b-5p过表达均促进SKOV3细胞凋亡,差异有统计学意义(P<0.05)并增加其放射敏感性,差异有统计学意义(P<0.05)。抑制miR-196b-5p表达可部分逆转抑制TUG1表达对SKOV3细胞促进凋亡和增强放射敏感性作用。结论 TUG1可负性调控miR-196b-5p的表达,抑制TUG1表达,可促进卵巢癌细胞凋亡,增加细胞放射敏感性。

Regulation of LncRNA TUG1 on apoptosis and radiosensitivity of the ovarian cancer cells by regulating miR-196b-5p expression

Objective To investigate the effect of LncRNA TUG1 on apoptosis and radiosensitivity of ovarian cancer cells and its potential mechanism by regulating miR-196 b-5 p expression.Methods The expression of TUG1 and miR-196 b-5 p in ovarian cancer cells SKOV3,HO8910 and OVCAR3 as well as normal ovarian epithelial cell HOSE were detected by Western blot and qRT-PCR.Possible target gene of TUG1 was verified with dual luciferase report gene assay.The SKOV3 cell lines transfected with si-TUG1,miR-196 b-5 p mimic and si-TUG1+miR-196 b-5 p were constructed,the cell apoptosis was determined by flow cytometry.Clonogenic survival assay and curve fitting were used to assess the changes in radiosensitivity of SKOV3 cells transfected with si-TUG1,miR-196 b-5 p mimic and si-TUG1+miR-196 b-5 p.Results Compared with normal ovarian epithelial cell HOSE,the expression of miR-196 b-5 p in ovarian cancer SKOV3,HO8910 and OVCAR3 was significantly decreased(P<0.05);and the expression of TUG1 was significantly increased(P<0.05).The inhibition of TUG1 or the over-expression of miR-196 b-5 p both promoted the apoptosis(P<0.05)of SKOV3 cells and enhanced its radiosensitivity(P<0.05).The effect of TUG1 inhibition on promoting apoptosis and increasing radiosensitivity of SKOV3 cells was partially reversed by the Inhibition of mir-196 b-5 p.Conclusion TUG1 negatively regulates the expression of mir-196 b-5 p,and the inhibition of TUG1 promotes apoptosis and increases radiosensitivity of ovarian cancer cells.