In Vitro Anticancer Potential of Jasione montana and Its Main Components against Human Amelanotic Melanoma Cells

Abstract

Jasione montana L. (Campanulaceae) is used in traditional Belarusian herbal medicine for sleep disorders in children, but the chemical composition and biological activity have not been investigated. In this study, the activities of J. montana extracts, their fractions and main compounds were evaluated in amelanotic melanoma C32 (CRL-1585) cells and normal fibroblasts (PCS-201-012). The extracts and fractions were analyzed using liquid chromatography–photodiode array detection–electrospray ionization–mass spectrometry (LC–PDA–ESI–MS/TOF) to characterize 25 compounds. Further, three major and known constituents, luteolin (22) and its derivatives such as 7-O-glucoside (12) and 7-O-sambubioside (9) were isolated and identified. The cytotoxic activities against fibroblasts and the amelanotic melanoma cell line were determined using the fixable viability stain (FVS) assay. The influence of diethyl ether (Et2O) fraction (JM4) and 22 on apoptosis induction was investigated using an annexin V binding assay. The obtained results showed significant cytotoxicity of JM4 and 22 with IC50 values of 119.7 ± 3.2 and 95.1 ± 7.2 μg/mL, respectively. The proapoptotic potential after 22 treatment in the C32 human amelanotic melanoma cell line was comparable to that of vinblastine sulfate (VLB), detecting 29.2 ± 3.0% apoptotic cells. Moreover, 22 displayed less necrotic potential against melanoma cells than VLB. In addition, the influences of JM4 and 22 on the dysfunction of the mitochondrial membrane potential (MMP), cell cycle and activity of caspases 3, 8, 9, and 10 were established. The effects of JM4 on MMP change (74.5 ± 3.0% of the cells showed a reduced MMP) corresponded to the results obtained from the annexin V binding assay and activation of caspase-9. JM4 and 22 displayed a significant impact on caspase-9 (40.9 ± 2.4% of the cells contained active caspase-9 after JM4 treatment and 16.6 ± 0.8% after incubation with 22) and the intrinsic (mitochondrial) apoptotic pathway. Moreover, studies have shown that JM4 and 22 affect the activation of external apoptosis pathways by inducing the caspase-8 and caspase-10 cascades. Thus, activation of caspase-3 and DNA damage via external and internal apoptotic pathways were observed after treatment with JM4 and 22. The obtained results suggest that J. montana extracts could be developed as new topical preparations with potential anticancer properties due to their promising cytotoxic and proapoptotic potential.

Wound Healing Properties of Jasione montana Extracts and Their Main Secondary Metabolites

“…Aboveground parts of J. montana were collected and identified as reported previously by Juszczak and coauthors. The plant material was pretreated and then processed to obtain extracts JM1 – JM3 and fractions JM4 – JM6 following a previously described procedure ( Juszczak et al, 2021 ).…”

“…The antioxidant activity of the JM1 – JM6 extracts and their main compounds ( 9 , 12 , and 22 ) against 2,2-diphenyl-1-picrylhydrazyl (DPPH) radicals was estimated according to the method previously described ( Ciganović et al, 2019 ) with some modifications. All extracts ( JM1 – JM6 ) and main components of the tested extracts ( 9 , 12 , and 22 ) were isolated and identified as previously described ( Juszczak et al, 2021 ). A total of 130 μL of sample solution at various concentrations was mixed with 70 μL of DPPH (0.2 mg/mL) in a 96-well plate.…”

“…To date, 5,7,3',4'-tetrahydroxyflavone (luteolin) ( 22 ), luteolin 7- O - β -D-glucoside (cynaroside) ( 12 ) and luteolin 7- O - β -D-xylosyl-(1-2)- β -D-glucoside (luteolin 7- O -sambubioside) ( 9 ) have been isolated. Moreover, the anticancer effect of J. montana has been documented in an in vitro model of skin melanoma ( Juszczak et al, 2021 ). As, from the pharmacological point of view, stimulation of proliferation, migration and differentiation of dermal fibroblasts, as well as anti-inflammatory or antioxidant effects, are critical aspects of the wound healing process, the aim of this study was to evaluate the effects of J. montana extracts and their main metabolites, such as luteolin, luteolin 7- O -glucoside and luteolin 7- O -sambubioside, on processes involved in wound healing.…”

Plumbagin Elicits Cell-Specific Cytotoxic Effects and Metabolic Responses in Melanoma Cells

“…Several natural phytochemical compounds with high anticancer potential are promising drugs in cancer therapy, including the treatment of melanoma [ 5 , 6 , 7 ]. Plumbagin (PLB), a naphthoquinone derivative mainly extracted from the plant, exhibits anticancer potential in different cancers, including inhibition of invasion and metastasis, induction of autophagy and apoptosis, and alteration of the cell cycle [ 8 , 9 , 10 , 11 ].…”

Flavonoids as tyrosinase inhibitors in in silico and in vitro models: basic framework of SAR using a statistical modelling approach

“…Compounds 5 (CAS number: 69097-99-0), 11 (CAS number: 480-40-0), 12 (CAS number: 520-36-5), 14 (CAS number: 3681-93-4), 20 (CAS number: 4261-42-1), 21 (CAS number: 491-71-4), 26 (CAS number: 520-18-3), 35 (CAS number: 6151-25-3), 39 (CAS number: 90-19-7), 40 (CAS number: 480-19-3), 42 (CAS number: 529-44-2) and 44 (CAS number: 446-72-0) were obtained from Carl Roth (Karlsruhe, Germany), 16 (CAS number: 480-44-4), 34 (CAS number: 489-32-7), and 43 (CAS number: 552-66-9) were obtained from Cayman Chemical (Ann Arbour, MI, USA). Flavonoids 1 , 2 , 28 , 33 12 , 13 13 , 15 , 31 , 38 14 , 17 15 , 18 , 19 16 , 7 , 8 , 9 17 , 22 , 23 , 24 , 25 18 , 27 , 32 19 , 29 , and 37 20 were isolated and identified in the Department of Pharmacognosy at the Medical University of Białystok. Compounds 3 (CAS number: 492-14-8), 4 (CAS number: 1036-49-3), and 10 (CAS number: 14813-19-5) were purchased from Biosynth Carbosynth (London, UK), 6 (CAS number: 520-26-3) and 41 (CAS number: 490-31-3) were purchased from Fluka Analysis (St. Louis, MO, USA), and 30 (CAS number: 22688-78-4) and 36 (CAS number: 22688-79-5) were obtained from Extrasynthese (Genay, France).…”