Effects of Annurca Flesh Apple Polyphenols in Human Thyroid Cancer Cell Lines

Abstract

Among natural macromolecules, the polyphenol extract from Annurca flesh (AFPE) apple could play a potential therapeutic role for a large spectrum of human cancer also by exerting antioxidant properties. Thyroid cancer is a common neoplasia in women, and it is in general responsive to treatments although patients may relapse and metastasize or therapy-related side effects could occur. In this study, we explored the effects of AFPE on papillary (TPC-1) and anaplastic (CAL62) thyroid cancer cell line proliferation and viability. We found that AFPE exposure induced a reduction of cell proliferation and cell viability in dose-dependent manner. The effect was associated with the reduction of phosphorylation of Rb protein. To study the mechanisms underlying the biological effects of AFPE treatment in thyroid cancer cells, we investigated the modulation of miRNA (miR) expression. We found that AFPE treatment increased the expression of the miR-141, miR-145, miR-200a-5p, miR-425, and miR-551b-5p. Additionally, since natural polyphenols could exert their beneficial effects through the antioxidant properties, we investigated this aspect, and we found that AFPE treatment reduced the production of reactive oxygen species (ROS) in CAL62 cells. Moreover, AFPE pretreatment protects against hydrogen peroxide-induced oxidative stress in thyroid cancer cell lines. Taken together, our findings suggest that AFPE, by acting at micromolar concentration in thyroid cancer cell lines, may be considered a promising adjuvant natural agent for thyroid cancer treatment approach.

Antineoplastic properties of polyphenols in TPC-1 human papillary thyroid carcinoma cell line: a systematic review

“…In contrast, only one study using pretreatment with AFPE showed a reduction in cell death induced by oxidative stress, underlying that polyphenols found specifically in this source may (alone or in combination) show a protective effect in this specific scenario (13). Finally, cell cycle and apoptosis regulators, inflammatory cytokines, inductors of cell growth, signaling pathways and altered mitochondrial membrane potentiality were also associated to the causing of cell death in TPC-1 lineage (6)(7)(8)(9)(10)(11)(12)(13)15). Therefore, it is possible to consider that many polyphenolic compounds have antineoplastic properties specifically in TPC-1 cell lineage.…”

“…(2022) applied it in a 24 hours exposition with a concentration of up to 250 μM/mL, resulting in a reduction of cell viability and promotion of cell cycle arrest analyzed by flow cytometry. Another concentration of AFPE (500 μM/mL) in a 20 hours exposition was evaluated by the DCFDA/H2DCFDA -Cellular ROS Assay Kit, obtaining a reduction of cell death induced by oxidative stress, contrasting with the findings described so far (13).…”

“…The induction of apoptosis, evaluated by flow cytometry (7,12), TUNEL (terminal deoxynucleotidyl transferase dUTP nick end labeling) assay (11), DNA extraction and fragmentation assay (8), and analysis through morphological examination in a fluorescence microscope (6) found DNA damage (8,15) and accumulation of ROS (6,15). In contrast, only one study using pretreatment with AFPE showed a reduction in cell death induced by oxidative stress, underlying that polyphenols found specifically in this source may (alone or in combination) show a protective effect in this specific scenario (13). Finally, cell cycle and apoptosis regulators, inflammatory cytokines, inductors of cell growth, signaling pathways and altered mitochondrial membrane potentiality were also associated to the causing of cell death in TPC-1 lineage (6)(7)(8)(9)(10)(11)(12)(13)15).…”

“…Western blottingEnhancement of Caspase3 and bax expression, and reduction of cyclin D1, c-Myc, survivin, and bcl-2 expression; suppression of PI3K/AKT pathway activation growth factor -VEGF and epidermal growth factor receptor -EGFR) and signaling pathways (mitogen activated protein kinase -MAPK/RAS-RAF-MEK extracellular signal-regulated kinases -ERK cascade and janus kinase -JAK/signal transducer and activator of transcription -STAT-3 pathway)(5)(6)(7)(8)(9)(10)(11)(12)(13)(14)(15).Accuracy of FNAB in thyroid nodule ≥4 cmArch Endocrinol Metab, 2023, v.67(6), 1-10, e000645.RT-qPCR Decrease bcl-2 expression and increase bax and caspases-3 expressionLiang and cols., 2020 Fisetin from 1 μM/mL to 30 μM/mL for 24 h MTT assay Promotion of a cytotoxic effect in a dose dependent mode15 μM/mL for 24 h Acronyms: MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide); bax (bcl2 associated X), bcl-xl (B-cell lymphoma); mcl-1 (induced myeloid leukemia cell differentiation); nrf2 (nuclear transcription factor -2); VEGF (vascular endothelial growth factor); TNFα (tumor necrosis factor α); ROS (Reactive Oxygen Species); PI3K/AKT (phosphatidylinositol-3-kinase/protein kinase B); AFPE (Annurca Flesh Apple); DCFDA/H2DCFDA -Cellular ROS (2',7'-dicholofluorescin diacetate and reactive oxygen species); EGCG (Epigallocatechin-3-gallate); EdU; (5-ethynyl-2'-deoxyuridine); MTS [3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(-4sulfophenyl)-2H-tetrazoluim, inner salt]; TUNEL (terminal deoxynucleotidyl transferase dUTP nick end labeling); PARP [poly(ADPribose) polymerase]; EGFR (epidermal growth factor receptor); RAS (rat sarcoma); RAF (rapidly accelerated fibrosarcoma); MEK (mitogen-activated protein kinase kinase); ERK (extracellular signalregulated kinase); PV (Prunella vulgaris); CCK-8 assay (cell counting kit-8); DNA (deoxyribonucleic acid); STAT3 (signal transducer and activator of transcription 3): JAK-1 (janus kinase-1); RT-qPCR (Reverse transcription-quantitative polymerase chain reaction).Accuracy of FNAB in thyroid nodule ≥4 cmArch Endocrinol Metab, 2023, v.67(6), 1-10, e000645.…”

Malus pumila Mill. cv Annurca apple extract might be therapeutically useful against oxidative stress and patterned hair loss

“…To date, this is the first study evaluating the potential ability of Annurca apple extract to counteract oxidative stress in HFDPCs. However, previous investigations have demonstrated the protective effect of Annurca against ROS on other cell lines, such as anaplastic thyroid cancer cells (CAL62) [ 36 ] and mammary epithelial cells (MCF10A) cells [ 37 ]. These results indicate that AT HAIR‐FUL AA ® can impede H 2 O 2 ‐treated HFDPC ROS accumulation and apoptosis and, therefore, may be used as a potential supplement for preventing hair loss induced by oxidative stress.…”