Background: Epithelial to mesenchymal transition (EMT) is a major determinant of cancer
metastasis and is closely linked with TGF-β1. Intracellular proteins, including E. Cadherin, N.
Cadherin and Vimentin are directly related to EMT that affect cell migration and adhesion; on the
other hand, non muscle myosin (NM) has a central role in cytokinesis, migration and adhesion.
Objective: We aimed to explore the association of EMT and metastasis with TGF-β1 through regulation
of non-muscle myosin II-A (NMII-A) and its interaction with Hexosamine Biosynthesis
Method: Protein expression changes were assessed by western blotting and immunofluorescent
staining while transcription level changes were assessed by qRT-PCR. EMT was assessed by phenotypic
analysis, wound healing, proliferation and transwell migration assay in vitro while in vivo
studies were conducted in BALB/c nude mice for lung orthotopic and tail vein metastasis models.
Results: We demonstrated that regulation of JNK/ P38/PI3K by TGF-β1 led to down expression of
NMII-A which promoted EMT and lung cancer metastasis. This down expression of NMII-A conversely
upregulated the expression of Core 2 N-acetyl Glucosaminyl Transferase mucin type
(C2GnT-M) and further facilitated up-regulation and down-regulation of N-acetylglucosaminyltransferase
(GnT) -V and -III respectively; moreover, NMII-A K.D cells showed 3 times
more tendency to migrate towards brain in vivo.
Conclusion: The study reports a novel pathway through which NMII-A negatively regulates EMT
and metastasis via up regulation of C2GnT-M, GnT-V and down expression of GnT-III. These findings
of lung cancer may further be required to study other cancer types.