Effect of miR-634 level on the growth of nasopharyngeal carcinoma cells and its mechanism
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摘要: 目的 研究鼻咽癌组织中miR-634水平及其对鼻咽癌细胞生长及MEK/ERK信号通路的影响,探讨miR-634在鼻咽癌中的作用机制。 方法 收集鼻咽癌组织标本60例和鼻咽部非癌组织标本60例。采用逆转录-聚合酶链反应(RT-PCR)法测定鼻咽癌组织和鼻咽癌细胞中miR-634水平。将人鼻咽癌CNE-1细胞分为空白对照组、阴性对照组和过表达miR-634组。采用MTT测定细胞增殖,采用流式细胞仪检测细胞凋亡,采用Western blotting测定鼻咽癌细胞细胞外调节蛋白激酶(ERK)、磷酸化细胞外调节蛋白激酶(p-ERK)、丝裂原活化蛋白激酶(MEK)、磷酸化丝裂原活化蛋白激酶(p-MEK)蛋白水平。 结果 鼻咽癌组织中miR-634水平低于癌旁组织(P<0.05)。鼻咽癌细胞中miR-634水平低于正常鼻咽部上皮细胞(P<0.05)。与空白对照组和阴性对照组比较,过表达miR-634组鼻咽癌细胞中miR-634水平和细胞凋亡率升高(P<0.05);p-ERK、p-MEK蛋白水平降低(P<0.05)。转染3 d和5 d,3组细胞OD值比较差异有统计学意义(P<0.05),与空白对照组和阴性对照组比较,过表达miR-634组细胞OD值降低(P<0.05)。 结论 鼻咽癌组织中miR-634水平降低,过表达miR-634可抑制鼻咽癌增殖、促进细胞凋亡,其机制可能与MEK/ERK信号通路有关。Abstract: Objective To investigate the level of miR-634 in nasopharyngeal carcinoma and its effects on the cell growth and MEK/ERK signaling pathway in nasopharyngeal carcinoma, and discuss the possible mechanism. Methods Total 60 specimens of nasopharyngeal carcinoma and 60 specimens of nasopharyngeal non-cancerous tissue were collected. The level of miR-634 in nasopharyngeal carcinoma tissues and nasopharyngeal carcinoma cells were determined by reverse transcription-polymerase chain reaction (RT-PCR). Human nasopharyngeal carcinoma CNE-1 cells were divided into blank control group, negative control group and overexpressed miR-634 group. The cell proliferation was measured by MTT. The cell apoptosis was detected by flow cytometry. The levels of extracellular regulated protein kinase (ERK), phosphorylated extracellular regulated protein kinase (p-ERK), mitogen-activated protein kinase (MEK) and phosphatiated mitogen-activated protein kinase (p-MEK) proteins in nasopharyngeal carcinoma cells were determined by Western blotting. Results The level of miR-634 in nasopharyngeal carcinoma tissues was lower than that in the adjacent tissues (P<0.05). The level of miR-634 in nasopharyngeal carcinoma cells was lower than that in normal nasopharyngeal epithelial cells (P<0.05). Compared with the blank control group and the negative control group, the level of miR-634 and the apoptosis rate in the overexpressed miR-634 group were increased (P<0.05); the levels of p-ERK, p-MEK mRNA and protein were decreased (P<0.05). After transfection for 3 d and 5 d, There was significant difference in OD values among the three groups (P<0.05), compared with the blank control group and the negative control group, the OD value of the overexpress miR-634 group was decreased (P<0.05). Conclusion The level of miR-634 is decreased in nasopharyngeal carcinoma, and overexpression of miR-634 can inhibit the proliferation of nasopharyngeal carcinoma and promote apoptosis. The mechanism may be related to MEK/ERK signaling pathway.
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