Abstract
BackgroundOsteopontin (OPN) is one of important molecular targets in cancer progression, metastasis as a calcium-binding, extracellular-matrix-associated protein of the small integrin-binding ligand and, N-linked glycoprotein. In the present study, anti-metastatic mechanism of ethanol extracts of Ocimum sanctum (EEOS) was elucidated on OPN enhanced metastasis in NCI-H460 non- small cell lung cancer cells.MethodsCell viability was measured by MTT assay. Adhesion and invasion assays were carried out to see that EEOS inhibited cell adhesion and invasion in OPN treated and non-treated NCI-H 460 cells. RT-PCR was used to determine the mRNA levels of uPA, uPAR, and EGFR.ResultsEEOS significantly inhibited cell adhesion and invasion in OPN treated and non treated NCI-H460 cells, though EEOS did not show any toxicity up to 200 μg/ml. EEOS effectively attenuated the expression of OPN and CD44 and also OPN activated the expression of CD44 in NCI-H460 cells. In addition, EEOS effectively suppressed the expression of phosphatidylinositide 3-kinases (PI3K) and cyclooxygenase 2 (COX-2) and the phosphorylation of Akt at protein level in OPN treated NCI-H460 cells. Also, EEOS significantly attenuated the expression of urokinase plasminogen activator (uPA), its receptor (uPAR) and epidermal growth factor receptor (EGFR) at mRNA level and reduced vascular endothelial growth factor (VEGF) production and MMP-9 activity in OPN treated NCI-H460 cells. Furthermore, PI3K/Akt inhibitor LY294002 enhanced anti-metastatic potential of EEOS to attenuate the expression of uPA and MMP-9 in OPN treated NCI-H 460 cells.ConclusionOverall, our findings suggest that anti-metastatic mechanism of EEOS is mediated by inhibition of PI3K/Akt in OPN treated NCI-H460 non-small cell lung cancer cells.
Highlights
Osteopontin (OPN) is one of important molecular targets in cancer progression, metastasis as a calcium-binding, extracellular-matrix-associated protein of the small integrin-binding ligand and, N-linked glycoprotein
In the present study, anti-metastatic mechanism of ethanol extracts of Ocimum sanctum (EEOS) was elucidated via cell viability assay, cell adhesion and invasion assays, ELISA for matrix metalloproteinase (MMP)-9, RT-PCR for urokinase plasminogen activator (uPA), uPAR and epidermal growth factor receptor (EGFR), Western blotting for osteopontin (OPN), and CD44, ELISA for vascular endothelial growth factor (VEGF) and phosphatidylinositide 3-kinases (PI3K)/Akt inhibitor LY294002 study in OPN treated NCI-H460 non-small cell lung cancer cells
Western blotting revealed that EEOS effectively attenuated the expression of OPN and CD44 and OPN activated CD44 at nontoxic concentrations in NCI-H460 cells
Summary
Osteopontin (OPN) is one of important molecular targets in cancer progression, metastasis as a calcium-binding, extracellular-matrix-associated protein of the small integrin-binding ligand and, N-linked glycoprotein. Anti-metastatic mechanism of ethanol extracts of Ocimum sanctum (EEOS) was elucidated on OPN enhanced metastasis in NCI-H460 non- small cell lung cancer cells. Though our group previously reported ethanol extract (95%) of OS (EEOS) exerts anti-metastatic activity via inhibition of MMP-9 and enhancement of antioxidant enzymes [32], the underlying anti-metastatic mechanism of EEOS still remains unclear. In the present study, anti-metastatic mechanism of EEOS was elucidated via cell viability assay, cell adhesion and invasion assays, ELISA for MMP-9, RT-PCR for uPA, uPAR and EGFR, Western blotting for osteopontin (OPN), and CD44, ELISA for VEGF and PI3K/Akt inhibitor LY294002 study in OPN treated NCI-H460 non-small cell lung cancer cells
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