MCF-7 Meme Kanseri Hücrelerinde Luteolin ile Birlikte Kuersetinin Apoptozun İndüksiyonu Yoluyla Sinerjistik Antikanser Aktivitesi

Year 2025, Volume: 12 Issue: 1, 136 - 144

Mehmet Kadir Erdoğan

Abstract

Kanser, artan insidansı ve mortalitesi ile büyük bir halk sağlığı sorunudur. Kanserin önlenmesi ve tedavisinde fitokimyasalların çarpıcı rolünün yadsınamazlığı yapılan çalışmalarla ortaya koyulmaktadır. Bu çalışmada kuersetin (QUR) ve luteolin (LTN) kombinasyonunun MCF-7 insan meme kanseri hücre hattı üzerindeki etkileri in vitro olarak incelendi. Sitotoksik aktivitenin belirlenmesinde WST-1 hücre sitotoksisitesi analizi kullanıldı. Koloni sağkalımı ve hücre göçü üzerindeki etkiler ise sırasıyla klonojenik analiz ve yara iyileşmesi analizi ile belirlendi. Apoptozun belirlenmesi için, AO/EB ikili boyama ve Kaspaz-3 ELISA analizi ile p53, Bcl-2, ve Bax ekspresyon düzeylerini ortaya koyan genomik analizler gerçekleştirildi. Analizler sonucunda özellikle QUR+LTN tedavisinin hücrelerde büyük bir sitotoksik aktivite sergilediği görüldü. Kombinasyon tedavisinin koloni sağkalımını baskıladığı ve hücre göçünü de önemli ölçüde inhibe ettiği belirlendi. Kantitatif analiz sonuçları QUR+LTN tedavisinin Kaspaz-3, p53, ve Bax'ı upregüle ederek, ve Bcl-2'yi ise downregüle ederek hücresel apoptozu tetiklediğini göstermektedir. Bu bulguların ileriki dönem in vitro ve in vivo analizlerle desteklenmesi, kombine fitokimyasal tedavi yaklaşımlarının kanser üzerindeki umut verici etkinliğinin ortaya konulmasına önemli katkılar sunabilir.

Keywords

Kanser, apoptoz, kuersetin, luteolin, MCF-7

Synergistic Anticancer Activity of Quercetin Combined with Luteolin in MCF-7 Breast Cancer Cells via Induction of Apoptosis

Abstract

Cancer is a major public health problem with its increasing incidence and mortality. The striking role of phytochemicals in the prevention and treatment of cancer is undeniable in studies. In this study, the effects of the combination of quercetin (QUR) and luteolin (LTN) on the MCF-7 human breast cancer cell line were investigated in vitro. WST-1 cell cytotoxicity assay was used to determine cytotoxic activity. The effects on colony survival and cell migration were determined by clonogenic assay and wound scratch assay, respectively. For the determination of apoptosis, AO/EB dual staining, and Caspase-3 ELISA analysis, and genomic analyses revealing p53, Bcl-2, and Bax expression levels were performed. As a result of the analyses, it was seen that especially QUR plus LTN treatment exhibited a great cytotoxic activity in cells. It was determined that the combination treatment suppressed colony survival and significantly inhibited cell migration. Quantitative analysis results show that QUR+LTN treatment triggers cellular apoptosis by upregulating Caspase-3, p53, and Bax, and downregulating Bcl-2. Supporting these findings with further in vitro and in vivo analyses may contribute significantly to revealing the promising efficacy of combined phytochemical treatment approaches on cancer.

Keywords

Cancer, apoptosis, quercetin, luteolin, MCF-7

References

• Albain, K. S., Nag, S. M., Calderillo-Ruiz, G., Jordaan, J. P., Llombart, A. C., Pluzanska, A., O'Shaughnessy, J. 2008. Gemcitabine plus paclitaxel versus paclitaxel monotherapy in patients with metastatic breast cancer and prior anthracycline treatment. Journal of Clinical Oncology, 26(24), 3950-3957.
• Bray, F., Laversanne, M., Sung, H., Ferlay, J., Siegel, R. L., Soerjomataram, I., Jemal, A. 2024. Global cancer statistics 2022: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries. CA: a cancer journal for clinicians, 74(3), 229-263.
• Chou, T. C. 2006. Theoretical basis, experimental design, and computerized simulation of synergism and antagonism in drug combination studies. Pharmacological reviews, 58(3), 621-681.
• Choudhari, A. S., Mandave, P. C., Deshpande, M., Ranjekar, P., Prakash, O. 2020. Phytochemicals in cancer treatment: From preclinical studies to clinical practice. Frontiers in pharmacology, 10, 1614.
• Erdogan, M. K., Agca, C. A., Askın, H. 2022. Quercetin and luteolin improve the anticancer effects of 5-fluorouracil in human colorectal adenocarcinoma in vitro model: A mechanistic insight. Nutrition and cancer, 74(2), 660-676.
• Fan, J. J., Hsu, W. H., Lee, K. H., Chen, K. C., Lin, C. W., Lee, Y. L. A., Cheng, C. H. 2019. Dietary flavonoids luteolin and quercetin inhibit migration and invasion of squamous carcinoma through reduction of Src/Stat3/S100A7 signaling. Antioxidants, 8(11), 557.
• Fantini, M., Benvenuto, M., Masuelli, L., Frajese, G. V., Tresoldi, I., Modesti, A., Bei, R. 2015. In vitro and in vivo antitumoral effects of combinations of polyphenols, or polyphenols and anticancer drugs: Perspectives on cancer treatment. International journal of molecular sciences, 16(5), 9236-9282.
• Ganai, S. A., Sheikh, F. A., Baba, Z. A., Mir, M. A., Mantoo, M. A., Yatoo, M. A. 2021. Anticancer activity of the plant flavonoid luteolin against preclinical models of various cancers and insights on different signalling mechanisms modulated. Phytotherapy Research, 35(7), 3509-3532.
• George, B. P., Chandran, R., Abrahamse, H. 2021. Role of phytochemicals in cancer chemoprevention: Insights. Antioxidants, 10(9), 1455
• Hanahan, D. 2022. Hallmarks of cancer: new dimensions. Cancer discovery, 12(1), 31-46.
• Kashyap, D., Mittal, S., Sak, K., Singhal, P., Tuli, H. S. 2016. Molecular mechanisms of action of quercetin in cancer: recent advances. Tumor Biology, 37, 12927-12939.
• Khdair, A., Chen, D., Patil, Y., Ma, L., Dou, Q. P., Shekhar, M. P., Panyam, J. 2010. Nanoparticle-mediated combination chemotherapy and photodynamic therapy overcomes tumor drug resistance. Journal of Controlled Release, 141(2), 137-144.
• Lamson, D. W., & Brignall, M. S. 2000. Antioxidants and cancer, part 3: quercetin. Alternative medicine review: a journal of clinical therapeutic, 5(3), 196-208.
• Lin, T. H., Hsu, W. H., Tsai, P. H., Huang, Y. T., Lin, C. W., Chen, K. C., Cheng, C. H. 2017. Dietary flavonoids, luteolin and quercetin, inhibit invasion of cervical cancer by reduction of UBE2S through epithelial–mesenchymal transition signaling. Food & Function, 8(4), 1558-1568.
• Livak, K. J., & Schmittgen, T. D. 2001. Analysis of relative gene expression data using real-time quantitative PCR and the 2− ΔΔCT method. Methods, 25(4), 402-408.
• Manson, M. M. 2003. Cancer prevention–the potential for diet to modulate molecular signalling. Trends in molecular medicine, 9(1), 11-18.
• Miean, K. H., & Mohamed, S. 2001. Flavonoid (myricetin, quercetin, kaempferol, luteolin, and apigenin) content of edible tropical plants. Journal of agricultural and food chemistry, 49(6), 3106-3112.
• Mokhtari, R. B., Homayouni, T. S., Baluch, N., Morgatskaya, E., Kumar, S., Das, B., Yeger, H. 2017. Combination therapy in combating cancer. Oncotarget, 8(23), 38022.
• Morales-Durán, N., León-Buitimea, A., Morones-Ramírez, J. R. 2024. Unraveling resistance mechanisms in combination therapy: A comprehensive review of recent advances and future directions. Heliyon.
• Nabavi, S. F., Braidy, N., Gortzi, O., Sobarzo-Sanchez, E., Daglia, M., Skalicka-Woźniak, K., Nabavi, S. M. 2015. Luteolin as an anti-inflammatory and neuroprotective agent: A brief review. Brain research bulletin, 119, 1-11.
• Nikanjam, M., Liu, S., Yang, J., Kurzrock, R. 2017. Dosing three‐drug combinations that include targeted anti‐cancer agents: analysis of 37,763 patients. The oncologist, 22(5), 576-584.
• Rizeq, B., Gupta, I., Ilesanmi, J., AlSafran, M., Rahman, M. M., Ouhtit, A. 2020. The power of phytochemicals combination in cancer chemoprevention. Journal of Cancer, 11(15), 4521.
• Sauter, E. R. 2020. Cancer prevention and treatment using combination therapy with natural compounds. Expert review of clinical pharmacology, 13(3), 265-285.
• Talib, W. H., Awajan, D., Hamed, R. A., Azzam, A. O., Mahmod, A. I., Al-Yasari, I. H. 2022. Combination anticancer therapies using selected phytochemicals. Molecules, 27(17), 5452.
• Yap, T. A., Omlin, A., De Bono, J. S. 2013. Development of therapeutic combinations targeting major cancer signaling pathways. Journal of clinical oncology, 31(12), 1592-1605.