Versican/PG-M is an extracellular matrix proteoglycan, expression of which is elevated in avariety of human tumors. The significance of this change is unclear. Here we show that versicanG3-containing fragments are present at high levels in human astrocytoma. Expression of aversican G3 construct in U87 astrocytoma cells enhances colony growth in soft agarose gel andtumor growth and blood vessel formation in nude mice. The G3-containing medium enhancesendothelial cell adhesion, proliferation, and migration. G3-expressing cells and tumors formedby these cells express increased levels of fibronectin and vascular endothelial growth factor(VEGF). Furthermore, the G3 domain directly binds to fibronectin and forms a complex togetherwith VEGF. In the presence of these three molecules, endothelial cell adhesion, proliferation,and migration were found to be significantly enhanced. Removal of the complex containing thesemolecules reverses these processes. Taken together, these findings implicate G3 as a modifier oftumor growth and angiogenesis and suggest a new avenue for development of anticancer andanti-angiogenic therapies based on targeting versican G3 fragments.

The chondroitin sulfate proteoglycan versican is one of the major extracellular components in the developing and adultbrain. Here, we show that isoforms of versican play different roles in neuronal differentiation and neurite outgrowth.Expression of versican V1 isoform in PC12 cells induced complete differentiation, whereas expression of V2 induced anaborted differentiation accompanied by apoptosis. V1 promoted neurite outgrowth of hippocampal neurons, but V2 failedto do so. V1 transfection enhanced expression of epidermal growth factor receptor and integrins, and facilitated sustainedextracellular signal-regulated kinase/MAPK phosphorylation. Blockade of the epidermal growth factor receptor, _1integrin, or Src significantly inhibited neuronal differentiation. Finally, we demonstrated that versican V1 isoform alsopromoted differentiation of neural stem cells into neurons. Our results have implications for understanding how versicanregulates neuronal development, function, and repair.

Versican is a large extracellular chondroitin sulfate proteoglycan that belongs to the family of lecticans. Alternativesplicing of versican generates at least four isoforms named V0, V1, V2, and V3. We show here that ectopic expression ofversican V1 isoform induced mesenchymal-epithelial transition (MET) in NIH3T3 fibroblasts, and inhibition of endogenousversican expression abolished the MET in metanephric mesenchyme. MET in NIH3T3 cells was demonstrated bymorphological changes and dramatic alterations in both membrane and cytoskeleton architecture. Molecular analysisshowed that V1 promoted a "switch" in cadherin expression from N-to E-cadherin, resulting in epithelial specificadhesion junctions. V1 expression reduced vimentin levels and induced expression of occludin, an epithelial-specificmarker, resulting in polarization of V1-transfected cells. Furthermore, an MSP (methylation-specific PCR) assay showedthat N-cadherin expression was suppressed through methylation of its DNA promoter. Exogenous expression of Ncadherinin V1-transfected cells reversed V1’s effect on cell aggregation. Reduction of E-cadherin expression by Snailtransfection and siRNA targeting E-cadherin abolished V1-induced morphological alteration. Transfection of an siRNAconstruct targeting versican also reversed the changed morphology induced by V1 expression. Silencing of endogenousversican prevented MET of metanephric mesenchyme. Taken together, our results demonstrate the involvement ofversican in MET: expression of versican is sufficient to induce MET in NIH3T3 fibroblasts and reduction of versicanexpression decreased MET in metanephric mesenchyme.

Versican is a large chondroitin sulfate proteoglycan belonging to the lectican family. Alternative splicing of versicangenerates at least four isoforms named V0, V1, V2, and V3. We have shown that the versican V1 isoform not only enhancedcell proliferation, but also modulated cell cycle progression and protected the cells from apoptosis. Futhermore, the V1isoform was able to not only activate proto-oncogene EGFR expression and modulate its downstream signaling pathway,but also induce p27 degradation and enhance CDK2 kinase activity. As well, the V1 isoform down-regulated theexpression of the proapoptotic protein Bad. By contrast, the V2 isoform exhibited opposite biological activities byinhibiting cell proliferation and down-regulated the expression of EGFR and cyclin A. Furthermore, V2 did not contributeapoptotic resistance to the cells. In light of these results, we are reporting opposite functions for the two versican isoformswhose expression is differentially regulated. Our studies suggest that the roles of these two isoforms are associated withthe subdomains CS and CS, respectively. These results were confirmed by silencing the expression of versican V1 withsmall interfering RNA (siRNA), which abolished V1-enhanced cell proliferation and V1-induced reduction of apoptosis.

Spontaneous loss of the Epstein-Barr virus (EBV) genome in the BL cell line Akata led to loss of tumorigenicityin SCID mice, suggesting an important oncogenic activity of EBV in B cells. We previously showed thatintroduction of the BARF1 gene into the human B-cell line Louckes induced a malignant transformation innewborn rats (M. X. Wei, J. C. Moulin, G. Decaussin, F. Berger, and T. Ooka, Cancer Res. 54:1843-1848, 1994).Since 1 to 2% of Akata cells expressed lytic antigens and expressed the BARF1 gene, we investigated whetherintroduction of the BARF1 gene into EBV-negative Akata cells can induce malignant transformation. Here weshow that BARF1-transfected, EBV-negative Akata cells activated Bcl2 expression and induced tumor formationwhen they were injected into SCID mice. In addition, when EBV-positive Akata cells expressing a low levelof BARF1 protein were injected into SCID mice, the expression of BARF1, as well as several lytic proteins, suchas EA-D, ZEBRA, and a 135-kDa DNA binding protein, increased in tumor cells while no latent LMP1 and lategp220-320 expression was observed in tumor cells. These observations suggest that the BARF1 gene may beinvolved in the conferral of tumorigenicity by EBV.