Formin-like 3 regulates RhoC/FAK pathway and actin assembly to promote cell invasion in colorectal carcinoma  

作　　者：Yuan-Feng Zeng Yi-Sheng Xiao Yong Liu Xiao-Jiang Luo Li-Dan Wen Qian Liu Min Chen 

机构地区：[1]Department of Pathology, Jiangxi Provincial People’s Hospital [2]Teaching and Researching Section of Morphology,College of Basic Medicine, Jiangxi University of Traditional Chinese Medicine [3]Department of General Surgery, Jiangxi Provincial People’s Hospital [4]Clinical Medical Sciences Institute, Jiangxi Provincial People’s Hospital

出　　处：《World Journal of Gastroenterology》2018年第34期3884-3897,共14页世界胃肠病学杂志（英文版）

基　　金：Supported by the National Natural Science Foundation of China,No.81201972;the China Postdoctoral Science Foundation,No.2013M531555;the Postdoctoral Science Foundation of Jiangxi province,No.2013KY44

摘　　要：AIM To clarify the underlying mechanism of formin-like 3(FMNL3)in the promotion of colorectal carcinoma(CRC)cell invasion.METHODS The in vitro biological function analyses of FMNL3 were performed by gain-and loss-of function approaches.Changes in the F-actin cytoskeleton were detected by the technologies of phalloidin-TRITC labeling and confocal microscopy.The signaling pathway mediated by FMNL3 was explored by western blot,gelatin zymograph assay,co-immunoprecipitation(co-IP),immunofluorescence colocalization,and glutathione S-transferase(GST)pulldown assay.RESULTS The in vitro experimental results showed that FMNL3 significantly promoted the proliferation,invasion,and migration of CRC cells(P<0.05 and P<0.01).Moreover,FMNL3regulated the remodeling of actin-based protrusions such as filopodia and lamellipodia in a RhoC-dependent manner.The western blot and gelatin zymograph assay results indicated that FMNL3 was involved in the RhoC/focal adhesion kinase(FAK)pathway and acted as an effector of RhoC to activate the downstream signaling of p-FAK as well as p-MAPK and p-AKT.This resulted in the increased expression of matrix metalloproteinase 2(MMP2),matrix metalloproteinase 9(MMP9)and vascular endothelial growth factor(VEGF),and the subsequent promotion of CRC cell invasion.The results of TAE226,U0126 or Ly294002 treatment confirmed an essential role of FMNL3 in activation of the RhoC/FAK pathway and the subsequent promotion of CRC invasion.Co-IP,colocalization and GST pull-down assays showed the direct interaction of FMNL3 with RhoC in vivo and in vitro.CONCLUSION FMNL3 regulates the RhoC/FAK signaling pathway and RhoC-dependent remodeling of actin-based protrusions to promote CRC invasion.AIM To clarify the underlying mechanism of formin-like 3(FMNL3)in the promotion of colorectal carcinoma(CRC)cell invasion.METHODS The in vitro biological function analyses of FMNL3 were performed by gain-and loss-of function approaches.Changes in the F-actin cytoskeleton were detected by the technologies of phalloidin-TRITC labeling and confocal microscopy.The signaling pathway mediated by FMNL3 was explored by western blot,gelatin zymograph assay,co-immunoprecipitation(co-IP),immunofluorescence colocalization,and glutathione S-transferase(GST)pulldown assay.RESULTS The in vitro experimental results showed that FMNL3 significantly promoted the proliferation,invasion,and migration of CRC cells(P<0.05 and P<0.01).Moreover,FMNL3regulated the remodeling of actin-based protrusions such as filopodia and lamellipodia in a RhoC-dependent manner.The western blot and gelatin zymograph assay results indicated that FMNL3 was involved in the RhoC/focal adhesion kinase(FAK)pathway and acted as an effector of RhoC to activate the downstream signaling of p-FAK as well as p-MAPK and p-AKT.This resulted in the increased expression of matrix metalloproteinase 2(MMP2),matrix metalloproteinase 9(MMP9)and vascular endothelial growth factor(VEGF),and the subsequent promotion of CRC cell invasion.The results of TAE226,U0126 or Ly294002 treatment confirmed an essential role of FMNL3 in activation of the RhoC/FAK pathway and the subsequent promotion of CRC invasion.Co-IP,colocalization and GST pull-down assays showed the direct interaction of FMNL3 with RhoC in vivo and in vitro.CONCLUSION FMNL3 regulates the RhoC/FAK signaling pathway and RhoC-dependent remodeling of actin-based protrusions to promote CRC invasion.

关 键 词：Formin-like 3 Colorectal carcinoma Invasion RhoC/FAK PATHWAY Actin assembly 

分 类 号：R57[医药卫生—消化系统]