Melittin Induced G1 Cell Cycle Arrest and Apoptosis in Chago-K1 Human Bronchogenic Carcinoma Cells and Inhibited the Differentiation of THP-1 Cells into Tumour- Associated Macrophages

Abstract

Background: Bronchogenic carcinoma (lung cancer) is one of the leading causes of death. Although many compounds isolated from natural products have been used to treat it, drug resistance is a serious problem, and alternative anti-cancer drugs are required. Here, melittin from Apis mellifera venom was used, and its effects on bronchogenic carcinoma cell proliferation and tumour-associated macrophage differentiation were evaluated. Methods: The half maximal inhibitory concentration (IC50) of melittin was measured by MTT. Cell death was observed by annexin V and propidium iodide (PI) co-staining followed by flow cytometry. Cell cycle arrest was revealed by PI staining and flow cytometry. To investigate the tumour microenvironment, differentiation of circulating monocytes (THP-1) into tumour-associated macrophages (TAMs) was assayed by sandwich-ELISA and interleukin (IL)-10 levels were determined. Cell proliferation and migration was observed by flat plate colony formation. Secretion of vascular endothelial growth factor (VEGF) was detected by ELISA. The change in expression levels of CatS, Bcl-2, and MADD was measured by quantitative RT-PCR. Results: Melittin was significantly more cytotoxic (p < 0.01) to human bronchogenic carcinoma cells (ChaGo-K1) than to the control human lung fibroblasts (Wi-38) cells. At 2.5 µM, melittin caused ChaGo-K1 cells to undergo apoptosis and cell cycle arrest at the G1 phase. The IL-10 levels showed that melittin significantly inhibited the differentiation of THP-1 cells into TAMs (p < 0.05) and reduced the number of colonies formed in the treated ChaGo-K1 cells compared to the untreated cells. However, melittin did not affect angiogenesis in ChaGo-K1 cells. Unlike MADD, Bcl-2 was up-regulated significantly (p < 0.05) in melittin-treated ChaGo-K1 cells. Conclusion: Melittin can be used as an alternative agent for lung cancer treatment because of its cytotoxicity against ChaGo-K1 cells and the inhibition of differentiation of THP-1 cells into TAMs.

Cytotoxic and Pro-Apoptotic Effects of a Sub-Toxic Concentration of Fluvastatin on OVCAR3 Ovarian Cancer Cells After its Optimized Formulation to Melittin Nano-Conjugates

“…As clearly depicted, FLV-MEL nano-conjugates treatment strongly inhibited the proliferation of OVCAR3 cells, measured by the reduction of G1→S transition as well as the inhibition of the transition from G2 to M phase. These effects could depend on the ability of MEL to suppress the proliferation of cancer cells ( Tipgomut et al, 2018 ; Ceremuga et al, 2020 ; Yao et al, 2020 ) which acts synergistically with FLV ( Hillyard et al, 2002 ; Garwood et al, 2010 ; Hwang et al, 2017 ) even when used at a sub-toxic concentration.…”

Use of Selected Carbon Nanoparticles as Melittin Carriers for MCF-7 and MDA-MB-231 Human Breast Cancer Cells

“…Also many studies show that MEL have a non-selective cytolytic effect, but it was proved that MEL selectively degrades cells' Ras oncogene expression which show overexpressed in 20-50 percent of breast cancers [32][33][34]. Furthermore, studies on normal and lung cancer cells have shown MEL caused lower cytotoxicity to the control human lung fibroblasts cells than to human lung cancer cells [35]. This study suggests that MEL in complexes could have a selective cytotoxic effect on cancer cells, but tests on healthy cells are needed.…”

Myomerger promotes fusion pore by elastic coupling between proximal membrane leaflets and hemifusion diaphragm

“…Fluorescent lipid Vybrant DiI, membrane-permeant Green CMFDA cell tracker and orange CMRA Cell Tracker were purchased from ThermoFisher Scientific (#V22885; #C7025 and #C34551, respectively). Penetratin and melittin were purchased from AnaSpec (catalogue # AS-64885) and Sigma-Aldrich (catalogue # M2272), respectively, and used in concentrations 0.25 and 0.5 μM for melittin; and 5 and 10 μM for penetratin, based on earlier studies 68 and 69 , respectively. Melittin and penetratin in concentrations exceeding 1 μM and 10 μM, respectively, caused apparent cytotoxicity.…”