Scorpion Venom Analgesic Peptide, BmK AGAP Inhibits Stemness, and Epithelial-Mesenchymal Transition by Down-Regulating PTX3 in Breast Cancer

Abstract

A scorpion peptide reported to exhibit both analgesic and antitumor activity in animal models may present as an alternative therapeutic agent for breast cancer. We aimed to investigate the effect of Buthus martensii Karsch antitumor-analgesic peptide (BmK AGAP) on breast cancer cell stemness and epithelial-mesenchymal transition (EMT). We treated MCF-7 and MDA-MB-231 cells with different concentrations of rBmK AGAP and observed that rBmK AGAP inhibited cancer cell stemness, epithelial-mesenchymal transition (EMT), migration, and invasion. Analysis by qPCR, ELISA, western blot, immunofluorescence staining, sphere formation, colony assay, transwell migration, and invasion assays demonstrated rBmK AGAP treatment decreased the expressions of Oct4, Sox2, N-cadherin, Snail, and increased the expression of E-cadherin. rBmK AGAP inhibited breast cancer cell stemness, EMT, migration, and invasion by down-regulating PTX3 through NF-κB and Wnt/β-catenin signaling Pathway in vitro and in vivo. Xenograft tumor model confirmed inhibition of tumor growth, stem-like features, and EMT by rBmK AGAP. Thus, rBmK AGAP is a potential therapeutic agent against breast cancer and related pain.

Levobupivacaine inhibits proliferation and promotes apoptosis of breast cancer cells by suppressing the PI3K/Akt/mTOR signalling pathway

“…Levobupivacaine at a concentration of 0, 1, 2 or 3 mM was used to treat MCF-7 and MDA-MB 231 cells plated in 96-well plates (1 × 10 4 cells/ well) and then incubated for 12, 24, or 48 h respectively in an incubator at the atmospheric condition of 37 °C with 5% CO 2 . The rest of the procedures used for the CCK-8 assay were the same as described elsewhere [18].…”

Levobupivacaine inhibits proliferation and promotes apoptosis of breast cancer cells by suppressing the PI3K/Akt/mTOR signalling pathway

“…Levobupivacaine at a concentration of 0, 1, 2 or 3mM was used to treat MCF-7 and MDA-MB 231 cells plated in 96-well plates (1×10⁴ cells/well) and then incubated for 12, 24, or 48 h respectively in an incubator at the atmospheric condition of 37 °C with 5 % CO . The rest of the procedures used for the CCK-8 assay were the same as described elsewhere [19].…”

“…The procedures used for the qPCR were the same as previously described [19]. The primers sequences were:…”

“…At the log phase of treated MCF-7 and MDA-MB 231 cells growth, we harvested the cells and then washed twice with ice-cold PBS. The rest of the procedures used for the western blot were the same as described elsewhere [19].…”

“…The MCF-7 and MDA-MBA-231 cells (5 × 104) that were pre-treated with different dose of Levobupivacaine (0, 1, 2 mM) for 24 h and resuspended in culture medium with the same concentrations of levobupivacaine were seeded onto the coated membrane in the upper chamber of the transwell (24well millicell cell culture insert, 12 mm diameter, 8 μm pores; Merck KGaA, #P18P01250, China). The procedures used for the Transwell assay were the same as previously describe [19] Values were expressed as the mean ± SD. Statistical analysis was performed with GraphPad Prism version 5.01(GraphPad Software, La Jolla, CA, US).…”