LncROR在促进鼻咽癌细胞增殖及上皮-间质转化作用中的研究

Study on the role of LncROR in promoting cell proliferation and epithelial mesenchymal transition of nasopharyngeal carcinoma

目的研究LncROR在鼻咽癌细胞增殖、抗凋亡及上皮-间质转化中的作用。方法收集2016年6月至2017年6月上海市浦东新区公利医院及长海医院经病理确诊的初治鼻咽癌患者及慢性鼻咽炎患者的病例标本各15例,分别采用实时荧光聚合酶链(qRT-PCR)方法检测lncROR的表达水平;以鼻咽癌细胞株CNE-2为研究对象,抑制lncROR表达后,CCK-8法检测CNE-2增殖能力变化,Western blot方法检测CNE-2凋亡相关蛋白(caspase-3, caspase-9)及上皮-间质转化相关蛋白(E-钙粘蛋白、β-连环蛋白、N-钙粘蛋白、波形蛋白)的表达变化。结果 LncROR在鼻咽癌组织及鼻咽癌细胞株中均有高表达,且在鼻咽癌细胞株CNE-2中表达明显高于其他细胞株;体外实验证实,抑制lncROR表达后,与转染对照组相比,细胞增殖能力明显减弱;活化的caspase-3,caspase-9蛋白表达升高;上皮标志蛋白(E-钙粘蛋白、β-连环蛋白)表达升高,间质标志蛋白(N-钙粘蛋白、波形蛋白)表达下降。结论 LncROR在鼻咽癌组织中高表达,并可以促进鼻咽癌细胞增殖、抗凋亡及上皮-间质转化。

Objective To study the role of LncROR in the proliferation, anti-apoptosis and epithelial-mesenchymal transition (EMT) of nasopharyngeal carcinoma cells. Methods The clinical samples were collected from patients pathologically diagnosed with nasopharyngeal carcinoma and chronic nasopharynx in the Gongli Hospital and Changhai Hospital affiliated to the Second Military Medical University. For each disease, 15 cases were collected during the time period from June 2016 to June 2017, respectively. The expression levels of lncROR of all the samples were detected by real-time fluorescence polymerase chain reaction (qRT-PCR). Furthermore, we also studied the function of LncROR in nasopharyngeal carcinoma cell line CNE-2. In particular, after the inhibition of lncROR expression, the proliferation of CNE-2 was characterized by CCK-8 assay, and the expression levels of apoptosis-related proteins (e.g., Caspase-3 and Caspase-9) and EMT-related proteins (E-cadherin,β-catenin, N-cadherin and Vimentin) were detected by Western blot. Results LncROR was highly expressed in both nasopharyngeal carcinoma and nasopharyngeal carcinoma cell lines, and the expression level in nasopharyngeal carcinoma cell line CNE-2 was significantly higher than that of other cell lines. In vitro experiments indicated that the cell proliferation ability was significantly reduced after the inhibition of lncROR when compared with the transfected control group;the expression of active caspase-3, caspase-9 protein was increased;the expression of epithelial marker protein (E-cadherin,β-catenin) was elevated;the expression level of interstitial marker protein (N-cadherin, vimentin) was down regulated. Conclusion LncROR is highly expressed in nasopharyngeal carcinoma, which can promote the proliferation,anti-apoptosis and epithelial-mesenchymal transformation of nasopharyngeal carcinoma cells.