Inhibitory effect of ethanol extract of Ocimum sanctum on osteopontin mediated metastasis of NCI-H460 non-small cell lung cancer cells

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.

Ethanolic extract of Ocimum sanctum leaves reduced invasion and matrix metalloproteinase activity of head and neck cancer cell lines

“…A previous study reported that 0.2 mg/ml EEOS demonstrated a significant cytotoxic effect on a lung cancer cell line (A549) and mouse Lewis lung carcinoma cells [19]. However, 0.2 mg/ml EEOS was non-toxic to NCI-H460 non-small cell lung cancer cells [18]. Our findings indicated that 0.05, 0.1, 0.2, and 0.4 mg/ml EEOS did not reduce HNSCC cell viability whereas, 0.8 mg/ml EEOS was toxic to the cell lines evaluated.…”

“…They found that EEOS inhibited cancer invasion and MMP-9 activity, but not that of MMP-2. Another group confirmed that EEOS reduced MMP-9 and urokinase plasminogen activator activity in non-small cell lung cancer cells (NCI-H460) by inhibiting the PI3K/Akt signalling pathway [18]. MMP-9 production in HNSCC cell is induced through various signalling pathways, including epidermal growth factor receptor (EGFR), mitogen-activated kinase (MAPK) and PI3K/Akt [26, 28].…”

“…The leaf extracts of O. sanctum have numerous medicinal effects such as anti-oxidant [12, 13], wound healing [14], anti-microbial [15] and anti-tumorigenic effects [11]. The ethanol extract of O. sanctum leaf (EEOS) has demonstrated anti-tumorigenic effects on several cancer types including gastric cancer [16], pancreatic cancer [17], non-small cell lung cancer [18], and lung cancers [19, 20]. EEOS exhibited a variety of therapeutic effects on tumor cells.…”

“…Moreover, EEOS inhibited cancer invasion and metastasis. EEOS reduced vascular endothelial growth factor production and MMP-9 activity in metastatic-induced NCI-H460 non-small cell lung cancer cells by inhibiting the phosphatidylinositol 3-kinase (PI3K)/Akt signaling pathway [18]. Similarly, when treated with EEOS, Lewis lung carcinoma cell MMP-9 activity was inhibited [20].…”

Interactions between osteopontin and vascular endothelial growth factor: Implications for cancer

“…They effectively attenuate the expression of OPN and CD44, as well as the OPN activated expression of CD44. Under treatment, the expression of urokinase plasminogen activator (uPA), its receptor uPAR, and the epidermal growth factor receptor are attenuated at the mRNA level and the production of VEGF and activity of MMP-9 are reduced [114]. The experimental anti-cancer drug tirapazamine is selectively activated by multiple reductases to form free radicals in hypoxic cells, thereby inducing DNA damage and cell death.…”