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

Expression of miR-214 in tongue squamous-cell carcinoma and its effects on cisplatin resistance

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 ICso 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 incrca3e （decrease） the sensitivity of tongue squamous-cell carcinoma cells to eisplatin, and this effect of miR-214 may be partly due to its regulation of Wnt/β-catenin signal pathway.