【摘要】 目的 探索miRNA-107通过RAP2B基因是否可以调控RhoA/ROCK通路对喉癌细胞上皮间质转化（Epithelial-mesenchymal transition,EMT）及增殖、侵袭能力产生影响探讨其作用机制。方法 将miR-107 mimic、si-RAP2B、miR-107 inhibitor、si-RAP2B+miR-107 inhibitor以及相对应的对照组转染至TU1和TU8细胞;CCK-8实验测定TU1和TU8细胞的增殖能力,Transwell试验测定TU1和TU8的侵袭能力,划痕实验测定TU1和TU8的迁移能力;Western blot分析RAP2B、RhoA、ROCK和EMT相关蛋白E-cadherin、Vimentin、N-cadherin的表达情况。结果miR-107过表达和沉默RAP2B均可降低TU1和TU8细胞增殖(F_TU1=14.652,F_TU8=13.248,P<0.01)、细胞侵袭(F_TU1=15.037,F_TU8=12.024,P<0.01)、细胞迁移能力(F_TU1=14.532,F_TU8=11.065,P<0.01)及Vimentin (t_TU1=8.28,t_TU8=8.16,P<0.01)、N-cadherin (t_TU1=7.63,t_TU8=8.04,P<0.01)表达,提高E-cadherin(t_TU1=8.69,t_TU8=7.63,P<0.01)的表达水平,抑制miR-107可逆转沉默RAP2B对TU1和TU8细胞的影响。结论miR-107在TU1和TU8组织低表达,其可能通过RAP2B基因调控RhoA/ROCK通路促进TU1和TU8细胞的侵袭、迁移及EMT。更多还原

【Abstract】 OBJECTIVE To explore whether miRNA-107 can regulate the RhoA/ROCK pathway through RAP2B gene on the epithelial mesenchymal transition,proliferation and invasion ability of laryngeal cancer cells,and to explore its mechanism.METHODS miR-107 mimic,si-RAP2B,miR-107 inhibitor,si-RAP2B+miR-107 inhibitor and corresponding controls were transfected into TU1 and TU8 cells.The proliferation ability of TU1 and TU8 cells was determined by CCK-8 assay,the invasion ability of TU1 and TU8 was determined by transwell assay,and the migration ability of TU1 and TU8 was determined by scratch assay.The expressions of RAP2B,RhoA,ROCK and EMT-related proteins E-cadherin,Vimentin and N-cadherin were analyzed by Western blot.RESULTS Both miR-107 overexpression RAP2B silence decreased the proliferation(F_TU1=14.652,F_TU8=13.248,P<0.01) and invasion(F_TU1=15.03 7,F_TU8=12.024,P<0.01) and migration ability(F_TU1=14.532,F_TU8=11.065,P<0.01) of TU1 and TU8 cells,and increased the Vimentin(t_TU1=8.28,t_TU8=8.16,P<0.01),N-cadherin(t_TU1=7.63,t_TU8=8.04,P<0.01) expres sion.Increasing the expression level of E-cadherin(t_TU1=8.69,t_TU8=7.63,P<0.01)and inhibiting miR-107 could reverse the effect of silenced RAP2B on TU1 and TU8 cells.CONCLUSION miR-107 is low expressed in TU1 and TU8 tissues respectively,which may promote the invasion,migration and epithelial mesenchymal transformation of TU1 and TU8 cells by regulating RhoA/ROCK pathway through RAP2B gene.更多还原