非肌肉肌球蛋白ⅡA通过c-jun氨基末端激酶信号途径调控胃癌细胞的迁移与侵袭

Nonmuscle myosin Ⅱ a regulates migration and invasion of gastric cancer cells via the c-jun N-terminal kinase/c-jun signaling pathway

目的 使用c-jun氨基末端激酶（JNK）/c-Jun信号通路的激活剂茴香霉素,观察对胃癌细胞迁移与侵袭的影响,探讨非肌肉肌球蛋白ⅡA（NMⅡA）是通过JNK/c-Jun信号通路米调控胃癌细胞的迁移与侵袭.方法 使用噻唑蓝（MTT）法分别测定0、5、10、20、40、80μg/L茴香霉素浓度对胃癌细胞的生长影响;使用RNA干扰（RNAi）干扰NMⅡA的表达,在胃癌细胞中加入JNK/c-Jun信号通路的激活剂茴香霉素,利用Western blot检测NMⅡA及JNK/c-Jun信号通路的相关蛋白表达水平.利用细胞划痕和Transwell实验来测定胃癌细胞的迁移和侵袭.结果 与对照组比较（0μg/L）,茴香霉素的浓度大于40 μg/L时,茴香霉素对胃癌细胞SGC-7901有明显抑制作用（P＜0.01）;Western blot结果显示,干扰NMⅡA的表达后,加入茴香霉素,可以增强JNK和c-Jun的磷酸化水平;划痕实验显示SGC-7901 Con对照组的组细胞划痕宽度由（600±25）μm降为（35 ±5）μm;而SGC-7901 RNAi干扰组的组细胞划痕宽度由之前的（610 ±30） μm降为（325 ±55） μm;当用茴香霉素处理SGC7-901 RNAi干扰组的细胞时,迁移距离由之前的（615 ±25）μm降为（90±35） μm;与干扰组比较,对照组和茴香霉素处理的实验组的迁移距离显著降低（P＜0.01）.Transwell实验结果茴香霉素处理的实验组,与干扰组比较,细胞迁移数目显著增多（P＜0.01）. 结论 在对胃癌细胞中NMⅡA通过JNK/c-Jun信号通路来调控胃癌细胞的迁移与侵袭.

Objective To observe the migration and invasion of gastric cancer cells treated with c-Jun N-terminal kinase （JNK）/c-Jun signaling pathways activator anisomycin,and to confirm the regulatory effects of nonmuscle myosin Ⅱ A （NM Ⅱ A） on migration and invasion of gastric cancer cells via the JNK/c-Jun signaling pathway.Methods Methyl thiazol tetrazolium （MTT） assay was used to detect the effect of different concentrations of anisomycin on the growth of gastric cancer cells.Western blotting was used to detect NM Ⅱ A and related protein expression level of JNK/c-Jun signaling pathways in gastric cancer cells treated with anisomycin.The effects of anisomycin on the invasion and migration rate of gastric cancer cell lines in vitro were measured using wound-healing and transwell assays.Results When the concentrations of anisomycin were more than 40 μg/L,anisomycin had obvious inhibitory effects on gastric cancer cells （P ＜ 0.01）.In gastric cancer cells with lower NM Ⅱ A expression,anisomycin could enhance phosphorylation level of JNK and c-Jun.Wound-healing results showed that the mnigration distance in SGC-7901 control group was decreased from （600 ±25） μm to （35 ±5） μm,and that in RNA interference（RNAi） group decreased from （610 ± 30） μm to （325 ± 55） μm.However,after treatment with anisomycin,the migration distance in RNAi group was decreased from （615 ± 25） μm to （90± 35） μm,and that in control group and RNAi group treated with anisomycin was significantly different from that in RNAi group without any treatment （P ＜ 0.01）.Transwell results revealed that the number of migrating cells was significantly increased in the experimental group treated with anisomycin as compared with RNAi group without any treatment （P ＜ 0.01）.Conclusion NM Ⅱ A regulates migration and invasion of gastric cancer cells via the JNK/c-Jun signaling pathway.