Meme kanserinde B13-klorokin uygulamasıyla in vitro tedavi araştırması

Abstract

Kanser hastalığı hala halk sağlığı için ciddi bir tehdit olmaya devam edmektedir. Meme kanseri tedavisi için yeni tedavi yaklaşımlarına ve yan etkisi düşük ajanlara ihtiyaç duyulmaktadır. Bunlara dayanarak, burada bir seramidaz inhibitörü (B13) ve bir otofaji inhibitörü (klorokin) içeren bir kombinasyonun insan meme kanseri hücre hattı üzerindeki sitotoksisitesinin araştırılması amaçlanmıştır. Antiproliferatif aktivite, Sulforhodamine B ve ATP canlılık deneyleri ile test edilmiştir. MCF-7 hücrelerinin ince yapısal ve morfolojik değişiklikleri ve apoptotik belirtileri için TEM ve konfokal mikroskopi teknikleri kullanılmıştır. Sonuçlar, kombinasyonun yüksek sitotoksik ve antiproliferatif aktivitelerinin yanı sıra apoptozu gösteren ince yapısal ve morfolojik değişikliklere neden olduğunu göstermiştir. B13+Klorokin kombinasyonunun MCF-7 hücrelerinde ölümü tetiklediği, antiproliferatif ve sitotoksik etkilere neden olduğu saptanmıştır. Sonuç olarak, yeni kombinasyonun antikanser bir ajan olarak ileri araştırmalar için iyi bir aday olduğu ortaya konulmuştur.

Keywords

• MCF-7
• kombine terapi
• sitotoksisite
• B13
• klorokin

Project Number

This study was supported by Anadolu University Scientific Research Project Unit with project number: 1901S002.

In vitro therapy investigation for breast cancer by B13-chloroquine application

Abstract

Cancer disease still remains to be strong treat for public health. New treatment approaches and agents with low side effects are needed for the treatment of breast cancer. Based on these, herein was aimed to investigate the cytotoxicity of a combination comprising a ceramidase inhibitor (B13) and an autophagy inhibitor (chloroquine) on human breast cancer cell line. The antiproliferative activity was tested by Sulforhodamine B and ATP viability assays. For ultrastructural and morphological changes and apoptotic signs of MCF-7 cells were used TEM and confocal microscopy techniques. Results showed the high cytotoxic and antiproliferative activities of the combination along with the ultrastructural and morphological changes indicating apoptosis. B13+Chloroquine combinations found to be effective on inducing cell death on MCF-7 cells and antiproliferative and cytotoxic effects on cells. Consequently, the new combination is suggested as good candidate for further investigations to be an anti-cancer agent.

Keywords

• MCF-7
• combination therapy
• cytotoxicity
• B13
• chloroquine

Supporting Institution

Anadolu University

Project Number

This study was supported by Anadolu University Scientific Research Project Unit with project number: 1901S002.

Thanks

This study was supported by Anadolu University Scientific Research Project Unit with project number: 1901S002.

References

1. References
2. [1] Mughal, T. A., Aslam, F., Yousaf, Z., Numrah, N., & Ping, C. L. (2020). In vitro cytotoxic activity of Zaleya pentandra L. extracts against the breast cancer adenocarcinoma cell line MCF-7. Journal of Pakistan Medical Association, 70, 35-41.
3. [2] Shen, S., Huang, L. S, Xiao, X. L., Zhu, X. F., Xiong, D. D., Cao, X. M., Wei, K. L., Chen, G., & Feng, Z. B. (2017). miR-204 regulates the biological behavior of breast cancer MCF-7 cells by directly targeting FOXA1. Oncology Reports, 38, 368-376.
4. [3] Nedaei, S. H., Akbari, H., & Yaghoubi, H. (2019). Cytotoxicity effects of green synthesized silver nanoparticles by using the extract of Tuber spp. on breast cancer (MCF-7) cells. Journal of Babol University Medical Sciences, 21, 242-48.
5. [4] Jardim, D. L., De Melo Gagliato, D., Nikanjam, M., Barkauskas, D. A., & Kurzrock, R. (2020). Efficacy and safety of anticancer drug combinations: a meta-analysis of randomized trials with a focus on immunotherapeutics and gene-targeted compounds. Oncoimmunology, 9, 1710052.
6. [5] Verbaanderd, C., Maes, H., Schaaf, M.B., Sukhatme, V. P., Pantziarka, P., Sukhatme, V., Agostinis, P., & Bouche, G. (2017). Repurposing drugs in oncology (redo)-chloroquine and hydroxychloroquine as anti-cancer agents. E. Cancer Medical Science. 23 (11) 781.
7. [6] Manic, G., Obrist, F., Kroemer, G., Vitale, I., & Galluzzi, L., (2014). Chloroquine and hydroxychloroquine for cancer therapy. Molecular and Cellular Oncology, 15, e29911.
8. [7] Al-Bari, M.A. (2015). Chloroquine analogues in drug discovery: new directions of uses, mechanisms of actions and toxic manifestations from malaria to multifarious diseases. Journal of Antimicrobial Chemotherapy, 70(6), 1608-1621.

9. [8] Selzner, M., Bielawska, A., Morse, M.A., Rüdiger, H. A., Sindram, D., Hannun, Y. A., & Clavien P. A., (2001). Induction of apoptotic cell death and prevention of tumor growth by ceramide analogues metastatic human colon cancer. Cancer Research, 61, 1233–1240.
10. [9] Raisova, M., Goltz, G., Bektas, M., Bielawska, A., Riebeling, C., Hossini, A. M., Eberle, J., Hannun, Y. A. Orfanos, C. E., & Geilen, C. C., (2002). Bcl-2 overexpression prevents apoptosis induced by ceramidase inhibitors in malignant melanoma and HaCaT keratinocytes. FEBS Letters, 516, 47–52.
11. [10] Samsel, L., Zaidel, G., Drumgoole, H. M., Jelovac, D., Drachenberg, C., Rhee, J., Brodie, A., Bielawska, A., & Smyth, M., (2004). The ceramide analog, B13, induces apoptosis in prostate cancer cell lines and inhibits tumor growth in prostate cancer xenografts. Prostate, 58, 382–393.
12. [11] Vejselova Sezer, C. (2021). Cytotoxic impacts of escin via inducing apoptosis and morphological changes on human prostate cancer cells. Biyolojik Çeşitlilik ve Koruma, 14 (1) , 82-87.
13. [12] Al-Attar, T. & Madihally, S. V. (2020). Recent advances in the combination delivery of drug for leukemia and other cancers. Expert Opinion in Drug Delivery, 17, 213-223.
14. [13] Fares, J., Kanojia, D., Rashidi, A., Ulasov, I., & Lesniak, M. S., (2020). Landscape of combination therapy trials in breast cancer brain metastasis. International Journal of Cancer, 147, 1939–1952.
15. [14] Schonewolf, C. A., Mehta, M., Schiff, D., Wu, H., Haffty, B. G., Karantza, V., & Jabbour S. K., (2014). Autophagy inhibition by chloroquine sensitizes HT-29 colorectal cancer cells to concurrent chemoradiation. World Journal of Gastrointestinal Oncology, 15, 74-82.
16. [15] Danial, N. N. & Korsmeyer, S. J. (2004). Cell death: critical control points. Cell, 116, 205-219.
17. [16] Kang, M. H. & Reynolds, C. P. (2009). Bcl-2 inhibitors: targeting mitochondrial apoptotic pathways in cancer therapy. Clinical Cancer Research,15(4), 1126-32.
18. [17] Liu, F., Shang, Y., & Chen, S. (2014). Chloroquine potentiates the anti-cancer effect of lidamycin on non-small cell lung cancer cells in vitro. Acta Pharmacologica Sinica, 35, 645–652.
19. [18] Solomon, V. R. & Lee, H. (2009). Chloroquine and its analogs: a new promise of an old drug for effective and safe cancer therapies. European Journal of Pharmacology, 625, 220–33.
20. [19] Fan, C., Wang, W., Zhao, B., Zhang, S., & Miao, J. (2006). Chloroquine inhibits cell growth and induces cell death in A549 lung cancer cells. Journal of Bioorganic and Medicinal Chemistry, 14(9), 3218-3222.
21. [20] Jiang, P. D., Zhao, Y. L., Deng, X. Q., Mao, Y. Q., Shi, W., Tang, Q. Q., Li, Z. G., Zheng, Y. Z., Yang, S. Y., & Wei, Y. W., (2010). Antitumor and antimetastatic activities of chloroquine diphosphate in a murine model of breast cancer. Biomedical Pharmacotherapy, 64, 609–614.