鼻咽癌患者中的miR-10b表达及调控机制研究

Study on the expression and regulation mechanism of miR-10b in patients with nasopharyngeal carcinoma

目的研究miR-10b在鼻咽癌患者活检组织中的表达及调控机制,探讨miR-10b与鼻咽癌细胞增殖和迁移的关系。方法取79例鼻咽癌初诊患者(观察组)和35例鼻咽正常黏膜组织标本(对照组),采用实时荧光定量PCR法检测两组miR-10b、LMP1和Twist1的表达情况,分析三者的相关性。Western法检测两组患者E-钙黏蛋白、β-连环蛋白、纤连蛋白、N-连环蛋白、波形蛋白和基质金属蛋白酶-9(MMP-9)的表达情况。结果miR-10b在鼻咽癌活检组织中的表达远高于鼻咽正常组织。Twist1以及EBV癌蛋白LMP1阳性组miR-10b过度表达(P<0.05)。鼻咽癌组织中上皮细胞标志物E-钙黏蛋白和β-连环蛋白的表达低于鼻咽正常组织,间充质细胞标志物纤连蛋白、N-钙黏蛋白、波形蛋白和MMP-9的表达高于鼻咽正常组织(P<0.05)。结论在鼻咽癌患者中,miR-10b基因通过LMP1、Twist1进行基因调控。过度表达的miR-10b促进鼻咽癌细胞的增殖和迁移,诱导上皮-间质转化(EMT),从而有可能成为治疗鼻咽癌的新靶点。

Objective To study the expression and regulation mechanism of miR-10 b in biopsy tissues of patients with nasopharyngeal carcinoma,and to explore the relationship between miR-10 b and proliferation and migration of nasopharyngeal carcinoma cells.Methods 79 nasopharyngeal carcinoma tissues(observation group)and 35 normal tissues of the nasopharynx(control group)were taken,the expression levels of miR-10 b and its upstream regulators LMP1 and Twist1 were examined by real-time fluorescence quantitative-PCR in the two groups,and their correlations were investigated.The expressions of E-cadherin,β-catenin,fibronectin,N-cadherin,vimentin and matrix metalloproteinase-9(MMP-9)in subjects of the two groups were detected by Western method.Results The expression of miR-10 b in nasopharyngeal carcinoma biopsy tissues was much higher than that in nasopharyngeal normal tissues.MiR-10 b was overexpressed in Twist1 and LMP1 positive group(P<0.05).The expressions of epithelial cell markers E-cadherin andβ-catenin in nasopharyngeal carcinoma tissues were lower than those in nasopharyngeal normal tissues(P<0.05).The expressions of mesenchymal cell markers fibronectin,N-cadherin,vimentin and MMP-9 in nasopharyngeal carcinoma tissues were higher than those in nasopharyngeal normal tissues(P<0.05).Conclusion miR-10 b is genetically adjusted by LMP1 and Twist1 in patients with nasopharyngeal carcinoma.The over-expressed miR-10 b promotes proliferation and migration of nasopharyngeal carcinoma cells and induces epithelial-mesenchymal transition(EMT),which may become a new target for the treatment of nasopharyngeal carcinoma.