miR-214在舌鳞癌组织中的表达及其对癌细胞顺铂敏感性的影响

摘要

目的：观察 miR-214在舌鳞癌组织中的表达情况及其对舌鳞癌细胞系顺铂敏感性的影响，并探讨其可能机制。方法采用实时荧光定量 PCR 检测50例份舌鳞癌组织、20例份癌旁组织、10例份正常舌黏膜组织及顺铂耐药舌鳞癌细胞系 TCA8113／DDP、亲本舌鳞癌细胞系 TCA8113中的 miR-214。将 miR-214模拟物及其阴性对照转染 TCA8113，将 miR-214阻遏物及其阴性对照转染 TCA8113／DDP，采用实时荧光定量 PCR 检测细胞中的 miR-214，CCK-8实验检测顺铂对细胞的半数抑制浓度（IC50），Western blot 检测细胞中的β-catenin、MDR1蛋白。结果舌鳞癌组织中 miR-214的相对表达量高于癌旁组织、正常舌黏膜组织（P 均＜0．01）。TCA8113／DDP 中 miR-214相对表达量高于 TCA8113细胞（P ＜0．01）。转染 miR-214模拟物 TCA8113细胞中的 miR-214、β-catenin 蛋白、MDR1蛋白表达水平及顺铂对其的 IC50均高于阴性对照细胞（P 均＜0．05）。转染 miR-214阻遏物 TCA8113／DDP 细胞中的 miR-214、β-catenin 蛋白、MDR1蛋白表达水平及顺铂对其的 IC50均低于阴性对照细胞（P 均＜0．05）。结论miR-214在舌鳞癌组织中高表达，下调（上调）miR-214的表达能够提高（降低）人舌鳞癌细胞对顺铂药物的敏感性，其机制可能与对 Wnt／β-catenin 信号通路的调控有关。%Objective To explore the expression of miR-214 in tongue squamous-cell carcinoma (TSCC)and its effects on cisplatin resistance in tongue squamous-cell carcinoma cells,and to explore the underlying mechanism.Methods The expression level of miR-214 was detected by real-time PCR in 50 samples of TSCC tissues,20 samples of para-carci-noma tissues,10 samples of normal tongue mucosa tissues,TCA8113 /DDP cells and TCA8113 cells.The TCA8113 cells were transfected with miR-214 mimics (miR-214 mimics group)or mimics-NC (mimics-NC group),and the TCA8113 /DDP were transfected with miR-214 inhibitor (miR-214 inhibitor group)or inhibitor-NC (inhibitor-NC group).The ex-pression level of miR-214 was detected by real-time PCR,the half maximal inhibitory concentration (IC50 )value of DDP by CCK8 and the expression level of β-catenin and multidrug resistance (MDR)1 protein by Western blotting.Results The expression of miR-214 in the TSCC tissues was significantly higher than that in para-carcinoma tissues and normal tongue mucosa tissues (all P <0.05).The expression level of miR-214 in the TCA8113 /DDP cells was significantly higher than that in TCA8113 cells (P <0.05).After TCA8113 cells were transfected with miR-214 mimics,the IC50 value and expression levels of miR-214,β-catenin and MDR1 in the miR-214 mimics group were significantly higher than those in the mimics-NC group (all P <0.05).After TCA8113 /DDP cells were transfected with miR-214 inhibitor,the IC50 value and expression levels of miR-214,β-catenin and MDR1 in the miR-214 inhibitor group were significantly lower than those in the inhibitor-NC group (all P <0.05).Conclusion The expression of miR-214 is highly expressed in the tongue squa-mous-cell carcinoma,the down-regulation(up-regulation)of miR-214 can effectively increase (decrease)the sensitivity of tongue squamous-cell carcinoma cells to cisplatin,and this effect of miR-214 may be partly due to its regulation of Wnt/β-catenin signal pathway.