TY - JOUR
T1 - Structure based design and synthesis of peptide inhibitor of human lox-12
T2 - In vitro and in vivo analysis of a novel therapeutic agent for breast cancer
AU - Singh, Abhay kumar
AU - Singh, Ratnakar
AU - Naz, Farhat
AU - Chauhan, Shyam Singh
AU - Dinda, Amit
AU - Shukla, Abhay Anand
AU - Gill, Kamaldeep
AU - Kapoor, Vaishali
AU - Dey, Sharmistha
N1 - Funding Information:
Authors acknowledge Council of Scientific and Industrial Research and Indian council of medical research, Government of India.
PY - 2012/2/23
Y1 - 2012/2/23
N2 - Human breast cancer cell proliferation involves a complex interaction between growth factors, steroid hormones and peptide hormones. The interaction of growth factors, such as epidermal growth factor (EGF), with their receptors on breast cancer cells can lead to the hydrolysis of phospholipids and release of fatty acid such as arachidonic acid, which can be further metabolized by cyclooxygenase (COX) and lipoxygenase (LOX) pathways to produce prostaglandins. The high concentration of prostaglandins has been associated with chronic inflammatory diseases and several types of human cancers. This is due to the over expression COX, LOX and other inflammatory enzymes. Ten peptides were designed and synthesized by solid phase peptide synthesis and analyzed in vitro for enzyme inhibition. Out of these peptides, YWCS had shown significant inhibitory effects. The dissociation constant (K D) was determined by surface plasmon resonance (SPR) analysis and was found to be 3.39×10 -8 M and 8.6×10 -8 M for YWCS and baicalein (positive control), respectively. The kinetic constant Ki was 72.45×10 -7 M as determined by kinetic assay. The peptide significantly reduced the cell viability of estrogen positive MCF-7 and estrogen negative MDA-MB-231 cell line with the half maximal concentration (IC 50) of 75 μM and 400 μM, respectively. The peptide also induced 49.8% and 20.8% apoptosis in breast cancer cells MCF-7 and MDA-MB-231, respectively. The YWCS was also found to be least hemolytic at a concentration of 358 μM. In vivo studies had shown that the peptide significantly inhibits tumor growth in mice (p<0.017). This peptide can be used as a lead compound and complement for ongoing efforts to develop differentiation therapies for breast cancer.
AB - Human breast cancer cell proliferation involves a complex interaction between growth factors, steroid hormones and peptide hormones. The interaction of growth factors, such as epidermal growth factor (EGF), with their receptors on breast cancer cells can lead to the hydrolysis of phospholipids and release of fatty acid such as arachidonic acid, which can be further metabolized by cyclooxygenase (COX) and lipoxygenase (LOX) pathways to produce prostaglandins. The high concentration of prostaglandins has been associated with chronic inflammatory diseases and several types of human cancers. This is due to the over expression COX, LOX and other inflammatory enzymes. Ten peptides were designed and synthesized by solid phase peptide synthesis and analyzed in vitro for enzyme inhibition. Out of these peptides, YWCS had shown significant inhibitory effects. The dissociation constant (K D) was determined by surface plasmon resonance (SPR) analysis and was found to be 3.39×10 -8 M and 8.6×10 -8 M for YWCS and baicalein (positive control), respectively. The kinetic constant Ki was 72.45×10 -7 M as determined by kinetic assay. The peptide significantly reduced the cell viability of estrogen positive MCF-7 and estrogen negative MDA-MB-231 cell line with the half maximal concentration (IC 50) of 75 μM and 400 μM, respectively. The peptide also induced 49.8% and 20.8% apoptosis in breast cancer cells MCF-7 and MDA-MB-231, respectively. The YWCS was also found to be least hemolytic at a concentration of 358 μM. In vivo studies had shown that the peptide significantly inhibits tumor growth in mice (p<0.017). This peptide can be used as a lead compound and complement for ongoing efforts to develop differentiation therapies for breast cancer.
UR - http://www.scopus.com/inward/record.url?scp=84863126444&partnerID=8YFLogxK
U2 - 10.1371/journal.pone.0032521
DO - 10.1371/journal.pone.0032521
M3 - Article
C2 - 22384268
AN - SCOPUS:84863126444
SN - 1932-6203
VL - 7
JO - PloS one
JF - PloS one
IS - 2
M1 - e32521
ER -