The effect of multiflora honey from Southeastern Anatolia on pro-inflammatory and pro-apoptotic pathways in MDA-MB-231 cell line
Yıl 2025,
Cilt: 50 Sayı: 3, 635 - 644, 30.09.2025
Dilan Erbaş
,
Aslı Özkızılcık Koçyiğit
,
Fikriye Fulya Kavak
,
Melek Pehlivan
,
Tülay Kılıçaslan Ayna
Öz
Purpose: Honey is increasingly recognized for its medicinal and nutritional benefits, including its anti-bacterial, anti-inflammatory, antioxidant, anti-ulcer, and anti-tumor properties. This study aimed to evaluate the effects of Anatolian honey (Southeast Anatolia) on the expression of pro-apoptotic genes and chemotactic molecules in breast cancer cells.
Materials and Methods: The IC50 concentration of Anatolian honey, determined to be 2.7% (w/v) at 48 hours, was used for treatment. Gene expression analysis was performed using the SYBR Green quantitative reverse transcription polymerase chain reaction (qRT-PCR) technique.
Results: The expression levels of the pro-apoptotic genes Bax and Caspase-3 increased by 6.05-fold and 2.01-fold, respectively, while the chemotactic molecule CCL2 exhibited a 6.45-fold upregulation. In contrast, CCL5 and CX3CR1 showed minimal changes in expression, with fold changes of 1.04 and 1.11, respectively. Anatolian honey promoted apoptosis by increasing Bax and Cas3 expression. Additionally, the elevated CCL2 expression suggests a potential role in recruiting monocytes and facilitating their differentiation into macrophages within the tumor microenvironment.
Conclusion: Anatolian honey exhibits pro-apoptotic and immunomodulatory effects in breast cancer cells by upregulating Bax, Cas3, and CCL2 expression. These findings highlight its potential as a complementary therapeutic agent.
Etik Beyan
Ethics Committee Approval is not necessary in this project, because this is an in vitro study including cell culture and it does not include any other human or animal materials.
Destekleyen Kurum
TUBITAK
Proje Numarası
1919B012101193
Kaynakça
-
Farcas CG, Dehelean C, Pinzaru IA, Mioc M, Socoliuc V, Moaca EA et al. Thermosensitive betulinic acid-loaded magnetoliposomes: a promising antitumor potential for highly aggressive human breast adenocarcinoma cells under hyperthermic conditions. Int J Nanomedicine. 2020;15:8175.
-
Arumugam A, Din Ibrahim M, Kntayya SB, Ain NM, Iori R, Galletti S et al. Induction of apoptosis by gluconasturtiin-isothiocyanate (GNST-ITC) in human hepatocarcinoma HepG2 cells and human breast adenocarcinoma MCF-7 cells. Molecules. 2020;25:1240.
-
Sanchez M, Barrere V, Treilleux I, Chopin N, Melodelima D. Development of a noninvasive HIFU treatment for breast adenocarcinomas using a toroidal transducer based on preliminary attenuation measurements. Ultrason Sonochem. 2021;115:106459.
-
Eteraf-Oskouei T, Najafi M. Uses of natural honey in cancer: an updated review. Adv Pharm Bull. 2022;12:248–61.
-
Qadirifard MS, Fathabadi A, Hajishah H, Gholami K, Abbasi M, Sami N et al. Anti-breast cancer potential of honey: a narrative review. OncoReview. 2022;12:5–15.
-
Yaacob NS, Nengsih A, Norazmi MN. Tualang honey promotes apoptotic cell death induced by tamoxifen in breast cancer cell lines. eCAM. 2013; 2013:989841.
-
Aryappalli P, Al-Qubaisi SS, Attoub S, George JA, Arafat K, Ramadi KB et al. The IL-6/STAT3 signaling pathway is an early target of Manuka honey-induced suppression of human breast cancer cells. Front Oncol. 2017;7:167.
-
Qiu J, Xu L, Zeng X, Wu H, Liang F, Lv Q et al. CCL5 mediates breast cancer metastasis and prognosis through CCR5/Treg cells. Front Oncol. 2022;12:972383.
-
Derossi DR, Amarante MK, Guembarovski RL, de Oliveira CEC, Suzuki KM, Watanabe MAE et al. CCL5 protein level: influence on breast cancer staging and lymph nodes commitment. Mol Biol Rep. 2019;46:6165–170.
-
Yoshimura T. The chemokine MCP-1 (CCL2) in the host interaction with cancer: a foe or ally? Cell Mol Immunol. 2018;15:335–45.
-
Rivas-Fuentes S, Salgado-Aguayo A, Arratia-Quijada J, Gorocica-Rosete P. Regulation and biological functions of the CX3CL1-CX3CR1 axis and its relevance in solid cancer: a mini-review. J Cancer. 2021;12:571.
-
Ponder KG, Boise LH. The prodomain of caspase-3 regulates its own removal and caspase activation. Cell Death Discov. 2019;5:1–10.
-
Cosentino K, Hertlein V, Jenner A, Dellmann T, Gojkovic M, Peña-Blanco A et al. The interplay between BAX and BAK tunes apoptotic pore growth to control mitochondrial-DNA-mediated inflammation. Mol Cell. 2022;82:933-49.
-
Rasulova K, Kavak DE, Kavak FF, Kandemir SI, Kızıldag S. The effect of multi oral honey on cytotoxicity and the mitochondrial apoptotic pathway in ovarian cancer cell. Med J Nutrition Metab. 2025;18:69-80.
-
Azimian H, Dayyani M, Bahreyni Toossi MT, Mahmoudi M. Bax/Bcl-2 expression ratio in prediction of response to breast cancer radiotherapy. Iran J Basic Med Sci. 2018;21:325.
-
Celesti G, Di Caro G, Bianchi P, Grizzi F, Marchesi F, Basso G et al. Early expression of the fractalkine Receptor CX3CR1 in pancreatic carcinogenesis. Br J Cancer. 2013;109:2424-33.
-
Yun Z, Chinnathambi A, Alharbi SA, Jin Z. Biosynthesis of gold nanoparticles using vetex negundo and evaluation of pro-apoptotic effect on human gastric cancer cell lines. J Photochem Photobiol B. 2020;203:111749.
-
Summer M, Ali S, Fiaz U, Tahir HM, Ijaz M, Mumtaz S et al. Therapeutic and immunomodulatory role of probiotics in breast cancer: a mechanistic review. Arch Microbiol. 2023;205:296.
-
Abel SDA, Dadhwal S, Gamble AB, Baird SK. Honey reduces the metastatic characteristics of prostate cancer cell lines by promoting a cross of adhesion. PeerJ. 2018;3:6:e5115.
-
Cianciosi D, Forbes-Hernández TY, Ansary J, Gil E, Amici A, Bompadre S et al. Phenolic compounds from mediterranean foods as nutraceutical tools for the prevention of cancer: the effect of honey polyphenols on colorectal cancer stem-like cells from spheroids. Food Chem. 2020;325:126881.
-
Ghramh HA, Alrumman SA, Ahmad I, Kalam A, Elbehairi SEI, Alfaify AM et al. Chemical characterization of honey and its effect (alone as well as with synthesized silver nanoparticles) on microbial pathogens’ and human cancer cell lines’ growth. Nutrients. 2023;15:684.
-
Martiniakova M, Kovacova V, Mondockova V, Zemanova N, Babikova M, Biro R et al. Honey: a promising therapeutic supplement for the prevention and management of osteoporosis and breast cancer. Antioxidants. 2023;12:567.
-
Erejuwa OO, Sulaiman SA, Ab Wahab MS. Effects of honey and its mechanisms of action on the development and progression of cancer. Molecules. 2014;19:2497.
-
Ahmed S, Othman NH. Honey as a potential natural anticancer agent: a review of its mechanisms. eCAM. 2013;2013:829070.
-
Aryappalli P, Shabbiri K, Masad RJ, Al-Marri RH, Haneefa SM, Mohamed YA et al. Inhibition of tyrosine-phosphorylated STAT3 in human breast and lung cancer cells by Manuka honey is mediated by selective antagonism of the IL-6 receptor. Int J Mol Sci. 2019;20:4340.
-
Seyhan MF, Yılmaz E, Timirci-Kahraman Ö, Saygılı N, Kısakesen Hİ, Eronat AP et al. Anatolian honey is not only sweet but can also protect from breast cancer: elixir for women from artemis to Present. IUBMB Life. 2017;69:677–88.
-
Nengsih A, Nik F, Yaacob S, Fauzi AN, Yaacob NS. Cell cycle and apoptosis pathway modulation by Tualang Honey in ER-dependent and -independent breast cancer cell lines. BMC Complement Altern Med. 2016;55:366–74.
-
Jiang M, Qi L, Li L, Li Y. The caspase-3/GSDME signal pathway as a switch between apoptosis and pyroptosis in Cancer. Cell Death Discov. 2020;6:1–11.
-
Chien SY, Wu YC, Chung JG, Yang JS, Lu HF, Tsou MF et al. Quercetin-induced apoptosis acts through mitochondrial- and caspase-3-dependent pathways in human breast cancer MDA-MB-231 Cells. Hum Exp Toxicol 2009;28:493–503.
-
Selina Darling-Reed P, Martin Luther King Jr Boulevard S, Bauer D, Mazzio E, Soliman KF et al. Diallyl disulfide inhibits TNFα-induced CCL2 release by MDA-MB-231 cells. Anticancer Res. 2014;34:2763-70.
-
An G, Wu F, Huang S, Feng L, Bai J, Gu S et al. Effects of CCL5 on the biological behavior of breast cancer and the mechanisms of its interaction with tumor associated macrophages. Oncol Rep. 2019;42:2499–2511.
-
Palacios-Arreola MI, Nava-Castro KE, Castro JI, García-Zepeda E, Carrero JC, Morales-Montor J. The role of chemokines in breast cancer pathology and its possible use as therapeutic targets. J Immunol Res. 2014;2014:849720
Güneydoğu Anadolu bölgesinde üretilen çiçek balının MDA-MB-231 hücre hattındaki pro-inflamatuar ve pro-apoptotik yolaklar üzerindeki etkisi
Yıl 2025,
Cilt: 50 Sayı: 3, 635 - 644, 30.09.2025
Dilan Erbaş
,
Aslı Özkızılcık Koçyiğit
,
Fikriye Fulya Kavak
,
Melek Pehlivan
,
Tülay Kılıçaslan Ayna
Öz
Amaç: Bal, anti-bakteriyel, anti-enflamatuvar, antioksidan, anti-ülser ve anti-tümör özellikleri gibi tıbbi ve besinsel faydalarıyla giderek daha fazla ilgi görmektedir. Bu çalışma, Anadolu balının meme kanserinde pro-apoptotik genler ve kemotaktik moleküller üzerindeki etkisini değerlendirmeyi amaçlamıştır.
Gereç ve Yöntem: Anadolu balının IC50 konsantrasyonu, 48. saatte %2.7 (ağırlık/hacim) olarak belirlenmiştir. Gen ekspresyon analizleri, SYBR Green yöntemi ile kantitatif gerçek zamanlı polimeraz zincir reaksiyonu (qRT-PCR) kullanılarak gerçekleştirilmiştir.
Bulgular: Pro-apoptotik genler olan Bax ve Kaspaz 3 (Cas3) ile kemotaktik molekül CCL2'nin ekspresyon seviyelerinde anlamlı bir artış gözlenmiştir. Buna karşın, CCL5 ve CX3CR1ekspresyon seviyelerinde herhangi bir değişiklik tespit edilmemiştir. Anadolu balı, Bax ve Cas3 gen ekspresyonunu artırarak apoptozu teşvik etmiştir. Ayrıca, CCL2 ekspresyonundaki artış, monositlerin tümör mikroçevresine toplanmasını ve makrofajlara farklılaşmasını kolaylaştırabileceğini göstermektedir.
Sonuç: Anadolu balı, meme kanserinde Bax, Cas3 ve CCL2 ekspresyonunu artırarak pro-apoptotik ve immün düzenleyici etkiler göstermektedir. Bu bulgular, Anadolu balının tamamlayıcı bir tedavi ajanı olarak potansiyelini ortaya koymaktadır.
Proje Numarası
1919B012101193
Kaynakça
-
Farcas CG, Dehelean C, Pinzaru IA, Mioc M, Socoliuc V, Moaca EA et al. Thermosensitive betulinic acid-loaded magnetoliposomes: a promising antitumor potential for highly aggressive human breast adenocarcinoma cells under hyperthermic conditions. Int J Nanomedicine. 2020;15:8175.
-
Arumugam A, Din Ibrahim M, Kntayya SB, Ain NM, Iori R, Galletti S et al. Induction of apoptosis by gluconasturtiin-isothiocyanate (GNST-ITC) in human hepatocarcinoma HepG2 cells and human breast adenocarcinoma MCF-7 cells. Molecules. 2020;25:1240.
-
Sanchez M, Barrere V, Treilleux I, Chopin N, Melodelima D. Development of a noninvasive HIFU treatment for breast adenocarcinomas using a toroidal transducer based on preliminary attenuation measurements. Ultrason Sonochem. 2021;115:106459.
-
Eteraf-Oskouei T, Najafi M. Uses of natural honey in cancer: an updated review. Adv Pharm Bull. 2022;12:248–61.
-
Qadirifard MS, Fathabadi A, Hajishah H, Gholami K, Abbasi M, Sami N et al. Anti-breast cancer potential of honey: a narrative review. OncoReview. 2022;12:5–15.
-
Yaacob NS, Nengsih A, Norazmi MN. Tualang honey promotes apoptotic cell death induced by tamoxifen in breast cancer cell lines. eCAM. 2013; 2013:989841.
-
Aryappalli P, Al-Qubaisi SS, Attoub S, George JA, Arafat K, Ramadi KB et al. The IL-6/STAT3 signaling pathway is an early target of Manuka honey-induced suppression of human breast cancer cells. Front Oncol. 2017;7:167.
-
Qiu J, Xu L, Zeng X, Wu H, Liang F, Lv Q et al. CCL5 mediates breast cancer metastasis and prognosis through CCR5/Treg cells. Front Oncol. 2022;12:972383.
-
Derossi DR, Amarante MK, Guembarovski RL, de Oliveira CEC, Suzuki KM, Watanabe MAE et al. CCL5 protein level: influence on breast cancer staging and lymph nodes commitment. Mol Biol Rep. 2019;46:6165–170.
-
Yoshimura T. The chemokine MCP-1 (CCL2) in the host interaction with cancer: a foe or ally? Cell Mol Immunol. 2018;15:335–45.
-
Rivas-Fuentes S, Salgado-Aguayo A, Arratia-Quijada J, Gorocica-Rosete P. Regulation and biological functions of the CX3CL1-CX3CR1 axis and its relevance in solid cancer: a mini-review. J Cancer. 2021;12:571.
-
Ponder KG, Boise LH. The prodomain of caspase-3 regulates its own removal and caspase activation. Cell Death Discov. 2019;5:1–10.
-
Cosentino K, Hertlein V, Jenner A, Dellmann T, Gojkovic M, Peña-Blanco A et al. The interplay between BAX and BAK tunes apoptotic pore growth to control mitochondrial-DNA-mediated inflammation. Mol Cell. 2022;82:933-49.
-
Rasulova K, Kavak DE, Kavak FF, Kandemir SI, Kızıldag S. The effect of multi oral honey on cytotoxicity and the mitochondrial apoptotic pathway in ovarian cancer cell. Med J Nutrition Metab. 2025;18:69-80.
-
Azimian H, Dayyani M, Bahreyni Toossi MT, Mahmoudi M. Bax/Bcl-2 expression ratio in prediction of response to breast cancer radiotherapy. Iran J Basic Med Sci. 2018;21:325.
-
Celesti G, Di Caro G, Bianchi P, Grizzi F, Marchesi F, Basso G et al. Early expression of the fractalkine Receptor CX3CR1 in pancreatic carcinogenesis. Br J Cancer. 2013;109:2424-33.
-
Yun Z, Chinnathambi A, Alharbi SA, Jin Z. Biosynthesis of gold nanoparticles using vetex negundo and evaluation of pro-apoptotic effect on human gastric cancer cell lines. J Photochem Photobiol B. 2020;203:111749.
-
Summer M, Ali S, Fiaz U, Tahir HM, Ijaz M, Mumtaz S et al. Therapeutic and immunomodulatory role of probiotics in breast cancer: a mechanistic review. Arch Microbiol. 2023;205:296.
-
Abel SDA, Dadhwal S, Gamble AB, Baird SK. Honey reduces the metastatic characteristics of prostate cancer cell lines by promoting a cross of adhesion. PeerJ. 2018;3:6:e5115.
-
Cianciosi D, Forbes-Hernández TY, Ansary J, Gil E, Amici A, Bompadre S et al. Phenolic compounds from mediterranean foods as nutraceutical tools for the prevention of cancer: the effect of honey polyphenols on colorectal cancer stem-like cells from spheroids. Food Chem. 2020;325:126881.
-
Ghramh HA, Alrumman SA, Ahmad I, Kalam A, Elbehairi SEI, Alfaify AM et al. Chemical characterization of honey and its effect (alone as well as with synthesized silver nanoparticles) on microbial pathogens’ and human cancer cell lines’ growth. Nutrients. 2023;15:684.
-
Martiniakova M, Kovacova V, Mondockova V, Zemanova N, Babikova M, Biro R et al. Honey: a promising therapeutic supplement for the prevention and management of osteoporosis and breast cancer. Antioxidants. 2023;12:567.
-
Erejuwa OO, Sulaiman SA, Ab Wahab MS. Effects of honey and its mechanisms of action on the development and progression of cancer. Molecules. 2014;19:2497.
-
Ahmed S, Othman NH. Honey as a potential natural anticancer agent: a review of its mechanisms. eCAM. 2013;2013:829070.
-
Aryappalli P, Shabbiri K, Masad RJ, Al-Marri RH, Haneefa SM, Mohamed YA et al. Inhibition of tyrosine-phosphorylated STAT3 in human breast and lung cancer cells by Manuka honey is mediated by selective antagonism of the IL-6 receptor. Int J Mol Sci. 2019;20:4340.
-
Seyhan MF, Yılmaz E, Timirci-Kahraman Ö, Saygılı N, Kısakesen Hİ, Eronat AP et al. Anatolian honey is not only sweet but can also protect from breast cancer: elixir for women from artemis to Present. IUBMB Life. 2017;69:677–88.
-
Nengsih A, Nik F, Yaacob S, Fauzi AN, Yaacob NS. Cell cycle and apoptosis pathway modulation by Tualang Honey in ER-dependent and -independent breast cancer cell lines. BMC Complement Altern Med. 2016;55:366–74.
-
Jiang M, Qi L, Li L, Li Y. The caspase-3/GSDME signal pathway as a switch between apoptosis and pyroptosis in Cancer. Cell Death Discov. 2020;6:1–11.
-
Chien SY, Wu YC, Chung JG, Yang JS, Lu HF, Tsou MF et al. Quercetin-induced apoptosis acts through mitochondrial- and caspase-3-dependent pathways in human breast cancer MDA-MB-231 Cells. Hum Exp Toxicol 2009;28:493–503.
-
Selina Darling-Reed P, Martin Luther King Jr Boulevard S, Bauer D, Mazzio E, Soliman KF et al. Diallyl disulfide inhibits TNFα-induced CCL2 release by MDA-MB-231 cells. Anticancer Res. 2014;34:2763-70.
-
An G, Wu F, Huang S, Feng L, Bai J, Gu S et al. Effects of CCL5 on the biological behavior of breast cancer and the mechanisms of its interaction with tumor associated macrophages. Oncol Rep. 2019;42:2499–2511.
-
Palacios-Arreola MI, Nava-Castro KE, Castro JI, García-Zepeda E, Carrero JC, Morales-Montor J. The role of chemokines in breast cancer pathology and its possible use as therapeutic targets. J Immunol Res. 2014;2014:849720