Phoyunnanin E inhibits migration of non-small cell lung cancer cells via suppression of epithelial-to-mesenchymal transition and integrin αv and integrin β3

Abstract

BackgroundThe conversion of the epithelial phenotype of cancer cells into cells with a mesenchymal phenotype-so-called epithelial–mesenchymal transition (EMT)-has been shown to enhance the capacity of the cells to disseminate throughout the body. EMT is therefore becoming a potential target for anti-cancer drug discovery. Here, we showed that phoyunnanin E, a compound isolated from Dendrobium venustum, possesses anti-migration activity and addressed its mechanism of action.MethodsThe cytotoxic and proliferative effects of phoyunnanin E on human non-small cell lung cancer-derived H460, H292, and A549 cells and human keratinocyte HaCaT cells were investigated by MTT assay. The effect of phoyunnanin E on EMT was evaluated by determining the colony formation and EMT markers. The migration and invasion of H460, H292, A549 and HaCaT cells was evaluated by wound healing assay and transwell invasion assay, respectively. EMT markers, integrins and migration-associated proteins were examined by western blot analysis.ResultsPhoyunnanin E at the concentrations of 5 and 10 μM, which are non-toxic to H460, H292, A549 and HaCaT cells showed good potential to inhibit the migratory activity of three types of human lung cancer cells. The anti-migration effect of phoyunnanin E was shown to relate to the suppressed EMT phenotypes, including growth in anchorage-independent condition, cell motility, and EMT-specific protein markers (N-cadherin, vimentin, slug, and snail). In addition to EMT suppression, we found that phoyunnanin E treatment with 5 and 10 μM could decrease the cellular level of integrin αv and integrin β3, these integrins are frequently up-regulated in highly metastatic tumor cells. We further characterized the regulatory proteins in cell migration and found that the cells treated with phoyunnanin E exhibited a significantly lower level of phosphorylated focal adhesion kinase (p-FAK) and phosphorylated ATP-dependent tyrosine kinase (p-AKT), and their downstream effectors (including Ras-related C3 botulinum (Rac-GTP); Cell division cycle 42 (Cdc42); and Ras homolog gene family, member A (Rho-GTP)) in comparison to those of the non-treated control.ConclusionsWe have determined for the first time that phoyunnanin E could inhibit the motility of lung cancer cells via the suppression of EMT and metastasis-related integrins. This new information could support further development of this compound for anti-metastasis approaches.

Natural Plants Compounds as Modulators of Epithelial-to-Mesenchymal Transition

“…Recent publications suggest that Phoyunnanin E decreased the E-cadherin to N-cadherin switch and reduced upregulation of mesenchymal markers such as vimentin and snail, as well as slug expression. Phoyunnanin-E has also been shown to inhibit migration and growth and promote EMT suppression, reduce migratory-associated integrins αv and β3, and suppress FAK/AKT cascade, which subsequently suppressed downstream migratory proteins in lung cancer cells (Petpiroon et al, 2017).…”

Antimetastatic Potential of Rhodomyrtone on Human Chondrosarcoma SW1353 Cells

“…Moreover, we further evaluated the downstream effectors of integrin including focal adhesion kinase (FAK), AKT, and p-AKT and small Rho GTPases family. There are many reports indicating that integrins initiate transmembrane signaling through activation of FAK and activate the downstream effectors such as AKT and Rho GTPases families for cancer cell migration [ 8 , 17 – 19 ]. Rho GTPases family including Ras, RhoA, Rac1, and Cdc42 are well known to regulate cytoskeleton remodeling, cell migration, malignant transformation, cell polarity, invasion, and metastasis [ 20 ].…”

“…Rho GTPases family including Ras, RhoA, Rac1, and Cdc42 are well known to regulate cytoskeleton remodeling, cell migration, malignant transformation, cell polarity, invasion, and metastasis [ 20 ]. Ras and Rac-1 regulate lamellipodia formation, Cdc42 stimulates plasma membrane protrusion filopodia, and RhoA promotes stress fiber formation [ 19 , 55 , 62 , 63 ]. Western blot analysis showed that rhodomyrtone significantly suppressed FAK expression and the downstream effectors including total AKT, phosphorylated AKT, Ras, RhoA, Rac1, and Cdc42 in SW1353 cell (Figures 6(c) and 6(f) ).…”

“…Overexpression of α 2 β 1, αvβ 1, αvβ 3, β 1, and β 1 integrin in human chondrosarcoma is associated with tumor progression, metastasis, and poor prognosis [ 5 , 8 , 15 , 16 ]. Integrins initiate transmembrane signaling through activation of the downstream effectors such as FAK, PI3K/AKT, and Rho GTPases family for cancer cell migration [ 8 , 17 – 19 ]. Small Rho GTPases including RhoA, Rac1, and Cdc42 are well known to regulate cell migration [ 20 ], and overexpression of these molecules in human cancer correlates with cancer progression [ 21 ].…”

Ovalitenone Inhibits the Migration of Lung Cancer Cells via the Suppression of AKT/mTOR and Epithelial-to-Mesenchymal Transition

“…For instance, gigantol, a bibenzyl compound extracted from Dendrobium draconis , has been shown to suppress EMT in non-small cell lung cancer H460 cells [ 22 ]. Petpiroon et al reported that phoyunnanin E inhibited the motility of lung cancer cells via the suppression of EMT and cancer metastasis-related integrins, such as integrin αv and integrin β3 [ 23 ]. Our previous results showed that ephemeranthol A, a natural compound isolated from D. infundibulum , also exhibited inhibitory effects on migration and EMT via suppression of the FAK-AKT signaling pathway [ 24 ].…”