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

Abstract

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 co-localization, and glutathione S-transferase (GST) pull-down 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, FMNL3 regulated 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, co-localization 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.

• Authors: Yuan-Feng Zeng, Yi-Sheng Xiao, Yong Liu, Xiao-Jiang Luo, Li-Dan Wen, Qian Liu, Min Chen
• Journal: World journal of gastroenterology (World J Gastroenterol) Vol. 24 Issue 34 Pg. 3884-3897 (Sep 14 2018) ISSN: 2219-2840 [Electronic] United States
• PMID: 30228782 (Publication Type: Journal Article)

Chemical References

• Actins
• Enzyme Inhibitors
• FMNL3 protein, human
• Formins
• Proteins
• RNA, Small Interfering
• Focal Adhesion Kinase 2
• PTK2B protein, human
• RHOC protein, human
• rhoC GTP-Binding Protein

Topics

• Actins (metabolism)
• Cell Adhesion (drug effects)
• Cell Line, Tumor
• Cell Movement (drug effects)
• Colorectal Neoplasms (pathology)
• Enzyme Inhibitors (pharmacology)
• Focal Adhesion Kinase 2 (genetics, metabolism)
• Formins
• HEK293 Cells
• Humans
• Neoplasm Invasiveness (pathology)
• Proteins (antagonists & inhibitors, genetics, metabolism)
• RNA, Small Interfering (metabolism)
• Signal Transduction (drug effects)
• rhoC GTP-Binding Protein (metabolism)