Limits...
Antibody and lectin target podoplanin to inhibit oral squamous carcinoma cell migration and viability by distinct mechanisms.

Ochoa-Alvarez JA, Krishnan H, Pastorino JG, Nevel E, Kephart D, Lee JJ, Retzbach EP, Shen Y, Fatahzadeh M, Baredes S, Kalyoussef E, Honma M, Adelson ME, Kaneko MK, Kato Y, Young MA, Deluca-Rapone L, Shienbaum AJ, Yin K, Jensen LD, Goldberg GS - Oncotarget (2015)

Bottom Line: Both reagents inhibited the migration of PDPN expressing OSCC cells at nanomolar concentrations before inhibiting cell viability at micromolar concentrations.In addition, both reagents induced mitochondrial membrane permeability transition to kill OSCC cells that express PDPN by caspase independent nonapoptotic necrosis.Taken together, these data suggest that antibodies and lectins may be utilized to combat OSCC and other cancers that express PDPN.

View Article: PubMed Central - PubMed

Affiliation: Departments of Molecular Biology, Cell Biology, and Pathology, School of Osteopathic Medicine, Rowan University, Stratford, NJ, USA.

ABSTRACT
Podoplanin (PDPN) is a unique transmembrane receptor that promotes tumor cell motility. Indeed, PDPN may serve as a chemotherapeutic target for primary and metastatic cancer cells, particularly oral squamous cell carcinoma (OSCC) cells that cause most oral cancers. Here, we studied how a monoclonal antibody (NZ-1) and lectin (MASL) that target PDPN affect human OSCC cell motility and viability. Both reagents inhibited the migration of PDPN expressing OSCC cells at nanomolar concentrations before inhibiting cell viability at micromolar concentrations. In addition, both reagents induced mitochondrial membrane permeability transition to kill OSCC cells that express PDPN by caspase independent nonapoptotic necrosis. Furthermore, MASL displayed a surprisingly robust ability to target PDPN on OSCC cells within minutes of exposure, and significantly inhibited human OSCC dissemination in zebrafish embryos. Moreover, we report that human OSCC cells formed tumors that expressed PDPN in mice, and induced PDPN expression in infiltrating host murine cancer associated fibroblasts. Taken together, these data suggest that antibodies and lectins may be utilized to combat OSCC and other cancers that express PDPN.

No MeSH data available.


Related in: MedlinePlus

NZ-1 and MASL do not induce caspase or PARP cleavage in OSCC cells(a) PARP, Caspase 8, and GAPDH were examined by Western blotting of protein from OSCC cells treated for 24 hours with 0 nM (control) or 2310 nM NZ-1 or MASL as indicated. (b) Signal was quantitated by image densitometry and shown as the percent of cleaved PARP and Caspase 8 compared to total PARP and Caspase 8, respectively (mean+SEM, n=3).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4496201&req=5

Figure 6: NZ-1 and MASL do not induce caspase or PARP cleavage in OSCC cells(a) PARP, Caspase 8, and GAPDH were examined by Western blotting of protein from OSCC cells treated for 24 hours with 0 nM (control) or 2310 nM NZ-1 or MASL as indicated. (b) Signal was quantitated by image densitometry and shown as the percent of cleaved PARP and Caspase 8 compared to total PARP and Caspase 8, respectively (mean+SEM, n=3).

Mentions: Effects on morphology suggest that NZ-1 and MASL utilize similar mechanisms to inhibit cell growth, but that cells can react differently to these effects. For example, both compounds induced some membrane blebbing of HSC-2 cells, but not HSC-4 cells. Effects of NZ-1 and MASL on caspase 8 and PARP cleavage were evaluated to further elucidate mechanisms behind their cytotoxicity. As shown in Figure 6, NZ-1 and MASL treated cells did not display significant levels of caspase 8 cleavage in response to these compounds. While some PARP cleavage was seen in HSC-2 cells treated with MASL, this was not found in NZ-1 treated HSC-2 cells or in the other cell lines treated with either reagent (Figure 6). These data suggest that caspase activation was not essential for NZ-1 or MASL toxicity.


Antibody and lectin target podoplanin to inhibit oral squamous carcinoma cell migration and viability by distinct mechanisms.

Ochoa-Alvarez JA, Krishnan H, Pastorino JG, Nevel E, Kephart D, Lee JJ, Retzbach EP, Shen Y, Fatahzadeh M, Baredes S, Kalyoussef E, Honma M, Adelson ME, Kaneko MK, Kato Y, Young MA, Deluca-Rapone L, Shienbaum AJ, Yin K, Jensen LD, Goldberg GS - Oncotarget (2015)

NZ-1 and MASL do not induce caspase or PARP cleavage in OSCC cells(a) PARP, Caspase 8, and GAPDH were examined by Western blotting of protein from OSCC cells treated for 24 hours with 0 nM (control) or 2310 nM NZ-1 or MASL as indicated. (b) Signal was quantitated by image densitometry and shown as the percent of cleaved PARP and Caspase 8 compared to total PARP and Caspase 8, respectively (mean+SEM, n=3).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4496201&req=5

Figure 6: NZ-1 and MASL do not induce caspase or PARP cleavage in OSCC cells(a) PARP, Caspase 8, and GAPDH were examined by Western blotting of protein from OSCC cells treated for 24 hours with 0 nM (control) or 2310 nM NZ-1 or MASL as indicated. (b) Signal was quantitated by image densitometry and shown as the percent of cleaved PARP and Caspase 8 compared to total PARP and Caspase 8, respectively (mean+SEM, n=3).
Mentions: Effects on morphology suggest that NZ-1 and MASL utilize similar mechanisms to inhibit cell growth, but that cells can react differently to these effects. For example, both compounds induced some membrane blebbing of HSC-2 cells, but not HSC-4 cells. Effects of NZ-1 and MASL on caspase 8 and PARP cleavage were evaluated to further elucidate mechanisms behind their cytotoxicity. As shown in Figure 6, NZ-1 and MASL treated cells did not display significant levels of caspase 8 cleavage in response to these compounds. While some PARP cleavage was seen in HSC-2 cells treated with MASL, this was not found in NZ-1 treated HSC-2 cells or in the other cell lines treated with either reagent (Figure 6). These data suggest that caspase activation was not essential for NZ-1 or MASL toxicity.

Bottom Line: Both reagents inhibited the migration of PDPN expressing OSCC cells at nanomolar concentrations before inhibiting cell viability at micromolar concentrations.In addition, both reagents induced mitochondrial membrane permeability transition to kill OSCC cells that express PDPN by caspase independent nonapoptotic necrosis.Taken together, these data suggest that antibodies and lectins may be utilized to combat OSCC and other cancers that express PDPN.

View Article: PubMed Central - PubMed

Affiliation: Departments of Molecular Biology, Cell Biology, and Pathology, School of Osteopathic Medicine, Rowan University, Stratford, NJ, USA.

ABSTRACT
Podoplanin (PDPN) is a unique transmembrane receptor that promotes tumor cell motility. Indeed, PDPN may serve as a chemotherapeutic target for primary and metastatic cancer cells, particularly oral squamous cell carcinoma (OSCC) cells that cause most oral cancers. Here, we studied how a monoclonal antibody (NZ-1) and lectin (MASL) that target PDPN affect human OSCC cell motility and viability. Both reagents inhibited the migration of PDPN expressing OSCC cells at nanomolar concentrations before inhibiting cell viability at micromolar concentrations. In addition, both reagents induced mitochondrial membrane permeability transition to kill OSCC cells that express PDPN by caspase independent nonapoptotic necrosis. Furthermore, MASL displayed a surprisingly robust ability to target PDPN on OSCC cells within minutes of exposure, and significantly inhibited human OSCC dissemination in zebrafish embryos. Moreover, we report that human OSCC cells formed tumors that expressed PDPN in mice, and induced PDPN expression in infiltrating host murine cancer associated fibroblasts. Taken together, these data suggest that antibodies and lectins may be utilized to combat OSCC and other cancers that express PDPN.

No MeSH data available.


Related in: MedlinePlus