Cleavage of galectin‐3 by matrix metalloproteases induces angiogenesis in breast cancer

Abstract

Galectin-3 cleavage is related to progression of human breast and prostate cancer and is partly responsible for tumor growth, angiogenesis and apoptosis resistance in mouse models. A functional polymorphism in galectin-3 gene, determining its susceptibility to cleavage by matrix metalloproteinases (MMPs)-2/-9 is related to racial disparity in breast cancer incidence in Asian and Caucasian women. The purpose of our study is to evaluate (i) if cleavage of galectin-3 could be related to angiogenesis during the progression of human breast cancer, (ii) the role of cleaved galectin-3 in induction of angiogenesis and (iii) determination of the galectin-3 domain responsible for induction of angiogenic response. Galectin-3 null breast cancer cells BT-459 were transfected with either cleavable full-length galectin-3 or its fragmented peptides. Chemotaxis, chemoinvasion, heterotypic aggregation, epithelial-endothelial cell interactions and angiogenesis were compared to noncleavable galectin-3. BT-549-H64 cells harboring cleavable galectin-3 exhibited increased chemotaxis, invasion and interactions with endothelial cells resulting in angiogenesis and 3D morphogenesis compared to BT-549-P64 cells harboring noncleavable galectin-3. BT-549-H64 cells induced increased migration and phosphorylation of focal adhesion kinase in migrating endothelial cells. Endothelial cells cocultured with BT-549 cells transfected with galectin-3 peptides indicate that amino acids 1–62 and 33–250 stimulate migration and morphogenesis of endothelial cells. Immunohistochemical analysis of blood vessel density and galectin-3 cleavage in a breast cancer progression tissue array support the in vitro findings. We conclude that the cleavage of the N terminus of galectin-3 followed by its release in the tumor microenvironment in part leads to breast cancer angiogenesis and progression.

The Great Escape; the Hallmarks of Resistance to Antiangiogenic Therapy

“…This corroborates previous findings by us and others showing that galectin-1 promotes tumor angiogenesis and is a target for antiangiogenic cancer therapy (Rabinovich et al, 2006;Thijssen et al, 2006Thijssen et al, , 2010Ito et al, 2011;Croci et al, 2012). Moreover, other members of the galectin protein family are expressed by EC (Thijssen et al, 2008) and have been associated with angiogenesis, including galectin-3 (Nangia- Makker et al, 2000Makker et al, , 2010Markowska et al, 2010), galectin-8 (Delgado et al, 2011), and galectin-9 (Heusschen et al, 2014;Thijssen and Griffioen, 2014). Similarly, as described by Croci et al (2014), their angioregulatory activity appears to involve glycandependent homo-and heterotypic receptor clustering as well as increasing the surface retention of receptors (Hsieh et al, 2008;Markowska et al, 2011;D'Haene et al, 2013).…”

Ocimum gratissimumretards breast cancer growth and progression and is a natural inhibitor of matrix metalloproteases

“…39 The presence of cleaved protein is associated with a more progressive tumor phenotype. 29,40 Incubation of galectin-3 with MMP-2 or MMP-9 in the presence of OG or its fractions resulted in reduced cleavage of galectin-3 by OG and HB, but not by HL suggesting the Collection) was maintained in Dulbecco's minimal essential medium (Invitrogen Corporation) containing 10% heatinactivated fetal calf serum (FCS), essential and nonessential amino acids (Invitrogen), vitamins and antibiotics (Mediatech Cellgro) as described.…”

“…Chemoinvasion was studied using a BD BioCoat Matrigel invasion chamber (BD Biosciences) according to manufacturer's instructions in the presence of 150 μg/mL OG, HB or HL as described. 29 The migrated cells were photographed using the Zeiss Axiovert 35 microscope supporting Olympus DP72 imaging system and counted using the Cellsens software MMP-2 and MMP-9 inhibitory activities were found to be enriched in methanol extractable HB fraction, while the chemotaxis inhibitory acivity was seen in the methanol non-extractable HL fraction.…”

“…Earlier we showed that active MMP-2 and MMP-9 cleave 31 kDa galectin-3 between Ala62-Tyr63 with resultant formation of a 22 kDa band. [29][30][31] Galectin-3 cleavage by MMP-2 was inhibited in a dose dependent fashion by OG and HB and to a lesser extent by HL. HB was most effective showing an inhibition of ~85% at a concentration of 450 μg/mL.…”

The Two Faces of Galectin-3: Roles in Various Pathological Conditions

“…It has been demonstrated that Galectin-3 favours a broad range of cancer cell activities, such as malignant cell transformation and tumour growth (62)(63)(64), cell adhesion, migration and invasion (65-69), anoikis resistance (70), apoptosis inhibition (71), (72) and angiogenesis (73).…”