Primary research

Stimulation of glioma cell motility by expression, proteolysis, and release of the L1 neural cell recognition molecule

Muhua Yang¹ , Shalini Adla¹ , Murali K Temburni^1,3 , Vivek P Patel¹ , Errin L Lagow¹ , Owen A Brady¹ , Jing Tian¹ , Magdy I Boulos² and Deni S Galileo¹

¹  Department of Biological Sciences, University of Delaware, Newark, DE 19716 USA

²  Helen F. Graham Cancer Center, Christiana Care Health System, Newark, DE 19713 USA

³  Department of Biology, Georgian Court University, Lakewood, NJ 08701 USA

author email corresponding author email

Cancer Cell International 2009, 9:27doi:10.1186/1475-2867-9-27

Published:	29 October 2009

Abstract

Background

Malignant glioma cells are particularly motile and can travel diffusely through the brain parenchyma, apparently without following anatomical structures to guide their migration. The neural adhesion/recognition protein L1 (L1CAM; CD171) has been implicated in contributing to stimulation of motility and metastasis of several non-neural cancer types. We explored the expression and function of L1 protein as a stimulator of glioma cell motility using human high-grade glioma surgical specimens and established rat and human glioma cell lines.

Results

L1 protein expression was found in 17 out of 18 human high-grade glioma surgical specimens by western blotting. L1 mRNA was found to be present in human U-87/LacZ and rat C6 and 9L glioma cell lines. The glioma cell lines were negative for surface full length L1 by flow cytometry and high resolution immunocytochemistry of live cells. However, fixed and permeablized cells exhibited positive staining as numerous intracellular puncta. Western blots of cell line extracts revealed L1 proteolysis into a large soluble ectodomain (~180 kDa) and a smaller transmembrane proteolytic fragment (~32 kDa). Exosomal vesicles released by the glioma cell lines were purified and contained both full-length L1 and the proteolyzed transmembrane fragment. Glioma cell lines expressed L1-binding αvβ5 integrin cell surface receptors. Quantitative time-lapse analyses showed that motility was reduced significantly in glioma cell lines by 1) infection with an antisense-L1 retroviral vector and 2) L1 ectodomain-binding antibodies.

Conclusion

Our novel results support a model of autocrine/paracrine stimulation of cell motility in glioma cells by a cleaved L1 ectodomain and/or released exosomal vesicles containing L1. This mechanism could explain the diffuse migratory behavior of high-grade glioma cancer cells within the brain.