Limits...
Recombinant disintegrin (r-Cam-dis) from Crotalus adamanteus inhibits adhesion of human pancreatic cancer cell lines to laminin-1 and vitronectin.

Suntravat M, Barret HS, Jurica CA, Lucena SE, Perez JC, Sánchez EE - J Venom Res (2015)

Bottom Line: The r-Cam-dis showed potent binding to αvβ3 integrin, but was moderate to weak with αvβ5, αvβ6, α2β1, and α6β1.Interestingly, the inhibition of r-Cam-dis on pancreatic cancer cell lines adhesion to laminin-1 was more effective than that to vitronectin.Based on our binding results to integrin receptors and previous adhesion studies using function-blocking monoclonal antibodies, it is suggested that r-Cam-dis could be inhibiting adhesion of pancreatic cancer cell lines through integrins α2β1, α6β1, αvβ5, and αvβ6.

View Article: PubMed Central - PubMed

Affiliation: National Natural Toxins Research Center, Texas A&M University-Kingsville, MSC 224, 975 West Avenue B, Kingsville, TX 78363, USA.

ABSTRACT
Pancreatic cancer is a malignant cancer common worldwide having poor prognosis, even when diagnosed at its early stage. Cell adhesion plays a critical role in cancer invasion and metastasis. Integrins are major mediators of cell adhesion and play an important role in invasion and metastatic growth of human pancreatic cancer cells. Snake disintegrins are the most potent ligands of several integrins and have potential therapeutic applications for cancers. We have previously cloned and expressed a new recombinant RGD-disintegrin from Crotalus adamanteus (r-Cam-dis). This recently published r-Cam-dis has an extra nine amino acids derived from the vector (SPGARGSEF) at the N-terminus end and has strong anti-platelet activity. However, this r-Cam-dis contains the contamination of the cleavage of the N-terminal end of the pET-43.1a cloning vector. In this study, we have cloned r-Cam-dis in a different cloning vector (pGEX-4T-1) showing five different amino acids (GSPEF) at the N-terminal part. This new r-Cam-dis was expressed and tested for inhibition of platelet aggregation, specific binding activity with seven different integrins, and inhibition of adhesion of three different pancreatic cancer cell lines on laminin-1 and vitronectin. The r-Cam-dis showed potent binding to αvβ3 integrin, but was moderate to weak with αvβ5, αvβ6, α2β1, and α6β1. Interestingly, the inhibition of r-Cam-dis on pancreatic cancer cell lines adhesion to laminin-1 was more effective than that to vitronectin. Based on our binding results to integrin receptors and previous adhesion studies using function-blocking monoclonal antibodies, it is suggested that r-Cam-dis could be inhibiting adhesion of pancreatic cancer cell lines through integrins α2β1, α6β1, αvβ5, and αvβ6.

No MeSH data available.


Related in: MedlinePlus

Inhibition of platelet aggregation using whole blood by r-Cam-dis. A Chronolog aggregometer was used to measure ADP-induced platelet aggregation by impedance. A total of 10μl of r-Cam-dis at varying concentrations was added to whole blood and incubated 1min at 37°C prior to adding 10μM of ADP. The error bars represent the standard deviation from three independent experiments with n=3.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4440708&req=5

Figure 2: Inhibition of platelet aggregation using whole blood by r-Cam-dis. A Chronolog aggregometer was used to measure ADP-induced platelet aggregation by impedance. A total of 10μl of r-Cam-dis at varying concentrations was added to whole blood and incubated 1min at 37°C prior to adding 10μM of ADP. The error bars represent the standard deviation from three independent experiments with n=3.

Mentions: r-Cam-dis was initially tested for the inhibition of ADP-induced platelet aggregation activity. The r-Cam-dis inhibited ADP-induced platelet aggregation in a dose-dependent manner with the IC50 value of 8.88nM (Figure 2).


Recombinant disintegrin (r-Cam-dis) from Crotalus adamanteus inhibits adhesion of human pancreatic cancer cell lines to laminin-1 and vitronectin.

Suntravat M, Barret HS, Jurica CA, Lucena SE, Perez JC, Sánchez EE - J Venom Res (2015)

Inhibition of platelet aggregation using whole blood by r-Cam-dis. A Chronolog aggregometer was used to measure ADP-induced platelet aggregation by impedance. A total of 10μl of r-Cam-dis at varying concentrations was added to whole blood and incubated 1min at 37°C prior to adding 10μM of ADP. The error bars represent the standard deviation from three independent experiments with n=3.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4440708&req=5

Figure 2: Inhibition of platelet aggregation using whole blood by r-Cam-dis. A Chronolog aggregometer was used to measure ADP-induced platelet aggregation by impedance. A total of 10μl of r-Cam-dis at varying concentrations was added to whole blood and incubated 1min at 37°C prior to adding 10μM of ADP. The error bars represent the standard deviation from three independent experiments with n=3.
Mentions: r-Cam-dis was initially tested for the inhibition of ADP-induced platelet aggregation activity. The r-Cam-dis inhibited ADP-induced platelet aggregation in a dose-dependent manner with the IC50 value of 8.88nM (Figure 2).

Bottom Line: The r-Cam-dis showed potent binding to αvβ3 integrin, but was moderate to weak with αvβ5, αvβ6, α2β1, and α6β1.Interestingly, the inhibition of r-Cam-dis on pancreatic cancer cell lines adhesion to laminin-1 was more effective than that to vitronectin.Based on our binding results to integrin receptors and previous adhesion studies using function-blocking monoclonal antibodies, it is suggested that r-Cam-dis could be inhibiting adhesion of pancreatic cancer cell lines through integrins α2β1, α6β1, αvβ5, and αvβ6.

View Article: PubMed Central - PubMed

Affiliation: National Natural Toxins Research Center, Texas A&M University-Kingsville, MSC 224, 975 West Avenue B, Kingsville, TX 78363, USA.

ABSTRACT
Pancreatic cancer is a malignant cancer common worldwide having poor prognosis, even when diagnosed at its early stage. Cell adhesion plays a critical role in cancer invasion and metastasis. Integrins are major mediators of cell adhesion and play an important role in invasion and metastatic growth of human pancreatic cancer cells. Snake disintegrins are the most potent ligands of several integrins and have potential therapeutic applications for cancers. We have previously cloned and expressed a new recombinant RGD-disintegrin from Crotalus adamanteus (r-Cam-dis). This recently published r-Cam-dis has an extra nine amino acids derived from the vector (SPGARGSEF) at the N-terminus end and has strong anti-platelet activity. However, this r-Cam-dis contains the contamination of the cleavage of the N-terminal end of the pET-43.1a cloning vector. In this study, we have cloned r-Cam-dis in a different cloning vector (pGEX-4T-1) showing five different amino acids (GSPEF) at the N-terminal part. This new r-Cam-dis was expressed and tested for inhibition of platelet aggregation, specific binding activity with seven different integrins, and inhibition of adhesion of three different pancreatic cancer cell lines on laminin-1 and vitronectin. The r-Cam-dis showed potent binding to αvβ3 integrin, but was moderate to weak with αvβ5, αvβ6, α2β1, and α6β1. Interestingly, the inhibition of r-Cam-dis on pancreatic cancer cell lines adhesion to laminin-1 was more effective than that to vitronectin. Based on our binding results to integrin receptors and previous adhesion studies using function-blocking monoclonal antibodies, it is suggested that r-Cam-dis could be inhibiting adhesion of pancreatic cancer cell lines through integrins α2β1, α6β1, αvβ5, and αvβ6.

No MeSH data available.


Related in: MedlinePlus