The expression of microRNA-145 in cervical cancer and its inhibitory effects on Wnt/β- catenin signaling pathway
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摘要:
目的 探讨microRNA-145在宫颈癌中表达的临床意义及其对Wnt/β-catenin信号通路的调控作用。 方法 收集2002年1月至2004年12月解放军464医院62例宫颈癌患者的组织标本, 采用实时荧光定量PCR 方法 检测microRNA-145表达并将其分为高表达组和低表达组, 分析其与临床病理参数的关系。将microRNA-145表达质粒和无义对照质粒分别转染入宫颈癌HeLa中, 分为过表达microRNA-145组和对照组, 采用细胞免疫荧光染色法分析细胞中β-catenin表达定位的变化, 双荧光素酶报告系统检测细胞中TCF/LEF转录活性及与Cateninδ-1的直接作用, 采用Western blot法检测microRNA-145对Cateninδ-1、C-MYC和Cy- clinD1表达的影响。 结果 microRNA-145低表达组患者的总生存期为(41.28±2.00)个月, 高表达组为(46.06±0.95)个月, 两者比较差异具有统计学意义(P < 0.05), 低表达组患者预后较差。过表达microRNA-145组HeLa细胞中的β-catenin主要分布于胞浆, 对照组主要位于细胞核和胞浆; 过表达microRNA-145组细胞中TCF/LEF转录活性受到抑制, 伴随着Cateninδ-1、C-MYC和Cy- clinD1表达下调, microRNA-145与Cateninδ-1可直接结合发挥作用。 结论 microRNA-145可通过与Cateninδ-1直接作用, 抑制Wnt/β-catenin信号通路转导, 从而发挥抑制宫颈癌演进的生物学功能。 -
关键词:
- microRNA-145 /
- 宫颈癌 /
- Wnt/β-catenin信号通路 /
- β-catenin /
- TCF/LEF /
- Cateninδ-1
Abstract:Objective To explore the clinic-pathological significance of microRNA-145 expression in human cervical cancer and its effects on Wnt/β-catenin signaling pathway. Methods Real-time PCR was used to detect the expression of microRNA-145 in 62 cervical cancer samples.The correlation between microRNA-145 expression and the clinic-pathological parameters and its prognostic significance was analyzed.MicroRNA-145-expressing plasmid and non-sense plasmid were transfected into human cervical cancer HeLa cells, assigned into overexpressed microRNA-145 group and control group.Immunofluorescence staining was performed to detect the expression location of β-catenin.Top Flash luciferase reporter assay was performed to investigate the effects of microRNA-145 on the transcriptional activity of TCF/LEF and direct interactions with Cateninδ-1.Western blot was used to detect the effects of microRNA- 145 on the expression of Cateninδ-1, C-MYC, and CyclinD1. Results The patients with low microRNA-145 expression showed poorer prognosis[(41.28±2.00) months vs.(46.06±0.95) months, P < 0.05].β-catenin immunofluorescence was distributed within the cytoplasm in the microRNA-145-overexpressed HeLa cells, but mainly within the nucleus and cytoplasm in the control cells.The luciferase reporter system indicated that the transcriptional activity of TCF/LEF was inhibited in the microRNA-145-overexpressed HeLa cells, and validated Cateninδ-1 was a target of miR-145.The expression of Cateninδ-1, C-MYC, and CyclinD1 was decreased in the microRNA-145- overexpressed HeLa cells. Conclusions microRNA-145 may inhibit cervical cancer progression via Cateninδ-1 and inhibit the Wnt/β- catenin signaling pathway. -
Key words:
- microRNA-145 /
- cervical cancer /
- Wnt/β-catenin signaling pathway /
- β-catenin /
- TCF/LEF /
- Cateninδ-1
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图 2 对照组及过表达microRNA-145组的HeLa细胞中β-catenin表达
A, D, G, J:对照组和过表达microRNA-145组中的β-catenin表达; B, E, H, K:对照组和过表达microRNA-145组中的DAPI表达; C, F, I, L:对照组和过表达microRNA-145组中的双通道β-catenin、DAPI共表达
表 1 宫颈癌组织中microRNA-145表达与各临床病理参数间的关系
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