miR-107通过RAP2B基因调控RhoA/ROCK通路对喉癌细胞上皮间质转化的机制研究  

Mechanism of miR-107 regulating RhoA/ROCK pathway through RAP2B gene on epithelial mesenchymal transformationof laryngeal carcinoma cells

作　　者：魏兴 刘海[2] 鄢斌成 蒋宗玲 段礼府[1] 马仁强[3] WEI Xing;LIU Hai;YAN Bincheng;JIANG Zongling;DUAN Lifu;MA Renqiang(Department of Otolaryngology Head and Neck Surgery,the First People's Hospital of Zigong City,Zigong,Sichuan,643000,China;Department of Otolaryngology Head and Neck Surgery,North Sichuan Medical College,Nanchong,Sichuan,637000,China;Department of Otolaryngology Head and Neck Surgery,the First Affiliated Hospital of Sun Yat-sen University,Guangzhou,Guangdong,510000,China)

机构地区：[1]自贡市第一人民医院耳鼻咽喉头颈外科,四川1自贡643000 [2]川北医学院耳鼻咽喉头颈外科,四川南充637000 [3]中山大学附属第一医院耳鼻咽喉头颈外科,广东广州510000

出　　处：《中国耳鼻咽喉头颈外科》2023年第9期551-556,共6页Chinese Archives of Otolaryngology-Head and Neck Surgery

基　　金：广东省自然科学基金项目(1614050000693)。

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(Epithelialmesenchymal transition,EMT)及增殖、侵袭能力产生影响并探讨其作用机制。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。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 and RAP2B silence decreased the proliferation(F_(TU1)=14.652,Frus=13.248,P<0.01)and invasion(F_(TU1)=15.037,Frus=12.024,P<0.01)and migration ability(F_(TU1)=14.532,Frus=11.065,P<0.01)of TU1 and TU8 cells,and increased the Vimentin(trui=8.28,trus=8.16,P<0.01),N-cadherin(trui=7.63,trus=8.04,P<0.01)expression.Increasing the expression level of E-cadherin(trui=8.69,trus=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.

关 键 词：微RNAS 癌 鳞状细胞 细胞增殖 细胞迁移分析 上皮间质转化 RAP2B RhoA/ROCK 

分 类 号：R739.65[医药卫生—肿瘤]