Glypican-1 Level Is Elevated in Extracellular Vesicles Released from MC38 Colon Adenocarcinoma Cells Overexpressing Snail

Abstract

The transcription factor Snail triggers epithelial-to-mesenchymal transition (EMT), endowing cancer cells with invasive properties during tumor progression. Extracellular vesicles (EVs) released from cancer cells at various stages of cancer progression are known to influence the tumor pre-metastatic niche and metastatic potential. The aim of this study was to analyze the effect of Snail on murine colon adenocarcinoma cells (MC38 line) and on the characteristics of their EVs. Stable clones of Snail-overexpressing MC38 cells were investigated in vitro versus Mock cells. Increased expression of matrix metalloproteinase MMP-14 and augmented activity of MMP-9 and -14 were observed in Snail-MC38 cells. There was no change in the transcriptomic profile of proteoglycans in Snail-MC38 cells; however, the protein level of Glypican-1 (GPC1) was enhanced in EVs released from those cells. Our finding that GPC1 protein level was enhanced in EVs released from MC38 cells that overexpressed Snail and were in an early EMT stage might explain the specificity of the GPC1 biomarker in colon cancer diagnosis. Further, our data suggest that Snail, by changing the level of GPC1 on EVs released by colon cancer cells, may affect the generation of a distant premetastatic niche and metastatic organotropism in colon adenocarcinoma.

Snail Overexpression Alters the microRNA Content of Extracellular Vesicles Released from HT29 Colorectal Cancer Cells and Activates Pro-Inflammatory State In Vivo

“…We reasoned that cancer cells at various EMT stages will release extracellular vesicles of different content and thus differently affect recipient cells. Changes in the protein levels in EVs released from cells stimulated by Snail were already observed [ 22 ]. As a rule, the cargo loaded into EVs reflects the status of cancer cells [ 23 ], but potential preferential packaging into EVs has also been suggested [ 11 ].…”

GPC1 promotes the growth and migration of colorectal cancer cells through regulating the TGF-β1/SMAD2 signaling pathway

“…Several studies demonstrated that GPC1 is involved in the formation and development of different types of tumors [ 20 , 21 ]. Experimental studies revealed that the expression of GPC1 was significantly increased in the extracellular vesicles released by the mouse MC38 CRC cell line [ 22 ]. In addition, TMT-MS methods to study crEV in patients with CRC found that GPC1 can be used as a biomarker for the detection of early CRC [ 23 ].…”

Advances in Biological Function and Clinical Application of Small Extracellular Vesicle Membrane Proteins

“…Overexpression of GPC1 activated epithelial-mesenchymal transition, which further enhanced CRC cell invasion and migration. Another study showed that GPC1 expression of sEVs released from mouse CRC cells was elevated and that high expression of GPC1 affected the generation of a distant premetastatic niche and metastatic organotropism in CRC (74). Furthermore, Frampton et al (75) discovered that silencing of GPC1 of pancreatic cancer cell-derived sEVs led to reduced tumor angiogenesis and an attenuation of mitogenic responses, which in turn slowed the growth of these tumor cells.…”

“…Overexpression of GPC1 activated epithelial-mesenchymal transition, which further enhanced CRC cell invasion and migration. Another study showed that GPC1 expression of sEVs released from mouse CRC cells was elevated and that high expression of GPC1 affected the generation of a distant premetastatic niche and metastatic organotropism in CRC ( 74 ). Furthermore, Frampton et al.…”