目的 探讨微小RNA（miR）-3651过表达通过介导核因子（NF）-κB信号通路抑制人舌癌细胞CAL27生长与侵袭的机制。 方法 将对数生长期CAL27细胞分为miR-3651 mimic组、mimic-NC组和对照组。采用qRT-PCR检测转染后细胞miR-3651水平，CCK-8检测细胞活力，Transwell检测细胞侵袭能力，划痕实验检测细胞迁移能力，光学显微镜下观察细胞上皮间质转化（epithelial-mesenchymal transition，EMT）的形成，Western blot检测E钙粘蛋白、N钙粘蛋白、波形蛋白、NF-κB p65和p-NF-κB p65蛋白表达。 结果 mimic-NC组与对照组细胞miR-3651水平、细胞活力、侵袭细胞数、细胞迁移率和细胞形态均无明显差异（P>0.05），E钙粘蛋白、N钙粘蛋白、波形蛋白和p-NF-κB p65/NF-κB p65蛋白表达均无明显差异（P>0.05）；与mimic-NC组相比，miR-3651 mimic组细胞miR-3651水平升高（P<0.05），培养的细胞第2、3、4 天的活力降低（P<0.05），侵袭细胞数和细胞迁移率降低（P<0.05），细胞E钙粘蛋白表达升高（P<0.05），N钙粘蛋白、波形蛋白和p-NF-κB p65/NF-κB p65蛋白表达下降（P<0.05），显微镜下观察到EMT形成减少。 结论 过表达miR-3651可以抑制CAL27细胞的生长和侵袭，其作用机制可能与抑制NF-κB信号通路磷酸化有关。
Abstract
Objective To explore the mechanism of microRNA (miR)-3651 overexpression on inhibiting the growth and invasion of human tongue cancer CAL27 cells by mediating nuclear factor (NF)-κB signaling pathway. Methods CAL27 cells in logarithmic growth phase were divided into a miR-3651 mimic group, a mimic-NC group and a control group. The level of miR-3651 in each group after transfection was detected by fluorescent quantitative real-time polymerase chain reaction (qRT-PCR). The cell vitality in each group was detected by cell counting kit-8 (CCK-8). The invasion ability of cells in each group was detected by Transwell. The migration ability of cells in each group was detected by scratch assay. The formation of epithelial-mesenchymal transition (EMT) in each group was observed under light microscope. The expression of E-cadherin, N-cadherin, Vimentin, NF-κB p65 and p-NF-κB p65 was detected by Western Blot. Results There was no statistically significant difference in miR-3651 level, activity of cells vitality, number of invasion cells, cells migration rate and cells morphology between mimic-NC group and control group (P>0.05), and there was no significant difference in the expression of E-cadherin, N-cadherin, Vimentin and p-NF-κB p65/NF-κB p65 proteins (P>0.05). Compared with the mimic-NC group, in the miR-3651 mimic group, miR-3651 level was significantly increased (P<0.05), cells vitality were significantly decreased at 2d, 3d, 4d after cell culture (P<0.05), number of invasion cells and cells migration rate were significantly decreased (P<0.05), expression of E-cadherin protein was significantly increased (P<0.05), expression of N-cadherin, Vimentin and p-NF-κB p65/NF-κB p65 proteins were significantly decreased (P<0.05), and EMT formation under microscope was decreased. Conclusions miR-3651 overexpression can inhibit the growth and invasion of CAL27 cells, and its action mechanism may be related to the inhibiting the phosphorylation of NF-κB signaling pathway.
关键词
微小RNA-3651； /
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核因子κB； /
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舌癌； /
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侵袭
Key words
microRNA-3651; /
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Nuclear factor κB; /
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Tongue cancer; /
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Invasion
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