局部麻醉剂通过丝裂原活化蛋白激酶途径诱导人甲状腺癌细胞凋亡

Local anesthetics induce apoptosis in human thyroid cancer cells via regulating mitogen-activated protein kinase pathway

目的探讨局部麻醉剂对人甲状腺癌细胞凋亡的影响及其作用机制。方法MTT法检测人甲状腺癌细胞经0 mM、1 mM、2 mM、4 mM、8 mM和16 mM的利多卡因及0 mM、0.2 mM、0.4 mM、0.8 mM、1.6 mM和3.2 mM的布比卡因分别干预24 h和48 h后的细胞活力流式细胞仪检测经4 mM和8 mM的利多卡因和0.8 mM和1.6 mM的布比卡因干预48 h后癌细胞的凋亡及线粒体膜电位;Western Blot检测凋亡相关因子及丝裂原活化蛋白激酶(MAPK)途径相关蛋白的表达;分别采用SP600125(JNK抑制剂)、PD98059(MEK抑制剂)、SB203580(p38 MAPK抑制剂)联合利多卡因或布比卡因对细胞进行处理,Western Blot检测凋亡相关蛋白的表达。结果利多卡因和布比卡因干预24 h和48 h均可呈剂量依赖式抑制人甲状腺癌细胞的生长。利多卡因和布比卡因干预48 h可降低癌细胞线粒体膜电位,增加Caspase-3和Bax的表达,降低Bcl-2的表达激活p38和JNK,抑制ERK的活性。MAPK信号通路抑制剂可降低利多卡因和布比卡因诱导Caspase-3和Bax的表达,增加Bcl-2的表达。结论局部麻醉剂利多卡因和布比卡因可诱导人甲状腺癌细胞生存抑制和凋亡,其机制与MAPK通路激活有关。

Objective To investigate the effect and the mechanism of local anesthetics on apoptosis of human thyroid cancer cells.Methods Human thyroid cancer cells were treated with 0,1 mM,2 mM,4 mM,8 mM and 16 mM of lidocaine and 0 mM,0.2 mM,0.4 mM,0.8 mM,1.6 mM and 3.2 mM of bupivacaine,respectively.After 24 h and 48 h of treatment,cell viability was analyzed by MTT assay.Flow cytometry was used to detect apoptosis and mitochondrial membrane potential of cancer cells after 48 h of treatment with 4 and 8 mM of lidocaine and 0.8 and1.6 mM of bupivacaine.Western Blot was used to detect the expressions of apoptosis-related factors and mitogen-activated protein kinase(MAPK) pathway-related proteins.The cells were treated with SP600125(JNK inhibitor),PD98059(MEK inhibitor),SB203580(p38 MAPK inhibitor) combined with lidocaine or bupivacaine,and the expression of apoptosis-related protein was detected by Western Blot.Results Both lidocaine and bupivacaine inhibited the growth of human thyroid cancer cells in a dose-dependent manner after 24 and 48 h of treatment.Lidocaine and bupivacaine reduced the mitochondrial membrane potential,increased the expressions of Caspase-3 and Bax,decreased the expression of Bel-2,activated p38 and JNK,and inhibited the activity of ERK.MAPK signaling pathway inhibitors reduced the expressions of Caspase-3 and Bax,and increased the expression of Bel-2 induced by lidocaine and bupivacaine.Conclusion Local anesthetics,such as lidocaine and bupivacaine,can induce survival inhibition and apoptosis in human thyroid cancer cells,and its mechanism is related to MAPK pathway activation.