Effective Targeting of Raf-1 and Its Associated Autophagy by Novel Extracted Peptide for Treating Breast Cancer Cells

Abstract

Breast cancer is one of the most common causes of death in women worldwide and has harmful influence on their psychological state during therapy. Multikinase inhibitors have become effective drugs for treating a variety of cancer diseases such as breast cancer. A purified short peptide (H-P) was isolated from the natural honey and tested for its potential regulatory role in breast cancer cells compared with the effectiveness of the anticancer drug, Sorafenib (SOR), using MCF-7, EFM-19, and MCF-10A cell lines. Furthermore, we investigated the direct connection between Raf-1 activation and cellular autophagy as potential targets of SOR and H-P extract using RNA interference. Interestingly, the treatment with H-P showed competitive regulation of phosphorylated Raf-1, MEK1/2, and matched autophagy-related LC3B without any detectable toxic effects in the non-tumorigenic epithelial cells. Unlike SOR, the regulation of Raf-1 protein and autophagic machinery by the novel H-P extract showed neglected levels of the released proinflammatory cytokine. This regulation of cytokine secretion by H-P resulted in decreasing the expression level of the transcription factor nuclear factor kappa-light-chain-enhancer of activated B cells (NF-kB) in treated cells. Moreover, the transfection of MCF-7 cells with small interference RNA (siRNA) antagonist Raf-1 expression markedly reduced the expression of LC3B, while it increased the expression of NF-kB1 and NF-kB2, indicating the potential cross-link between Raf-1, autophagy, and NF-kB effector. Collectively, these findings suggest that H-P-mediated Raf-1, MEK1/2, LC3B, and NF-kB provide a novel and efficacious multikinase inhibitor for treating breast cancer without detectable cytotoxic effects.

Potential Anticancer Activity of the Furanocoumarin Derivative Xanthotoxin Isolated from Ammi majus L. Fruits: In Vitro and In Silico Studies

“…It has been reported recently that autophagy induces pro-survival signals that confer protection to cancer. Molecules inhibiting autophagy are important for effective targeting of cancer cells [22]. To assess the effect of xanthotoxin on autophagy, HepG2 cells were cultured with 6.9 µg/mL concentration of xanthotoxin for 48 h, and acridine orange flow cytometry flux was measured to determine autophagy.…”

Anticancer Properties of N,N-dibenzylasparagine as an Asparagine (Asp) analog, Using Colon Cancer Caco-2 Cell Line

“…Finally, 100 µl SDS-HCl solution was added to each well in the plate which then was incubated for 4 hrs at 37ºC. Cell viability was then assessed based on the amount of converted water-soluble MTT to an insoluble formazan which is then solubilized and determined by optical density at 570 nm (El-Fadl et al, 2021).…”

Anti-cancer effect of some Egyptian natural plants extraction against prostate cancer cells

“…The content of garlic, cinnamon and liquorice liquids was quantitatively measured by high performance liquid chromatography (HPLC) (El-Fadl et al, 2021). The used HPLC (Agilent 1100) is composed of a quaternary pump and a UV/V as a detector.…”

“…The PCR reaction system conatined 10 µl SYBR green, 0.25 µl RNase inhibitor (25 U/µl), 0.2 μM of each primer, 2 μL of synthesized cDNA, and nuclease-free water up to a final volume of 25 μL. The PCR was used at conditions of 94°C for 5 min, 35 cycles (94°C for 30 sec, 60°C for 15 sec, 72°C for 30 sec) (Khalil et al, 2019;El-Fadl et al, 2021).…”