Investigation of the Effects of Piperlongumine and Doxorubicin Combined Treatment on Cell Death via PTEN in HeLa Cells

Yıl 2024, , 1 - 11, 30.04.2024

Gonca Güzelel , Hande Akalan , Türker Bilgen , Duygu Yaşar Şirin

https://doi.org/10.53433/yyufbed.1258515

Öz

Doxorubicin (Dox), which is used in treating many types of cancer including cervix cancer nevertheless, its effect alone is low especially in recurrent cases. Therefore, investigating of agents that can increase the impact of Dox continues. The aim of the present study is to answer the question: Can Piperlongumine (PL) a natural alkaloid cause an increase in the efficacy of Dox in the HeLa cell line? In this study, the effects of Dox and PL on cell viability by MTT and Acridine orange/propidium iodide staining, and expression levels of the PTEN (Phosphatase and tensin homolog 10) gene by Real-Time PCR and Western-Blot were evaluated in HeLa cells. It was determined that PL combined with Dox increased cell death and suppressed cell proliferation. The PTEN gene expression was decreased in all experimental groups, but the PTEN protein phosphorylation increased in cultures treated with PL and when Dox/PL was combined. The fact that PL application increases the activation of PTEN, which is a tumor suppressor. This indicates that it can be used to increase the effectiveness of Dox in the treatment.

Anahtar Kelimeler

Doxorubicin, Piperlongumine, PTEN

Destekleyen Kurum

Tekirdağ Namık Kemal University Scientific Research Projects Unit

Proje Numarası

NKUBAP.01.YL.20.267

HeLa Hücrelerinde Piperlongumin ve Doksorubisin Kombine Tedavisinin PTEN Yoluyla Hücre Ölümü Üzerine Etkilerinin Araştırılması

Öz

Doksorubisin (Dox) birçok kanser türünün yanı sıra serviks kanseri tedavisinde de kullanılmaktadır ancak özellikle tekrarlayan vakalarda tek başına etkisi düşüktür. Bu nedenle Dox etkisini artırabilecek ajanların araştırılması devam etmektedir. Bu çalışmada amaç: Doğal bir alkaloid olan Piperlongumine (PL), HeLa hücre hattında Dox etkinliğini artırabilir mi? sorusuna cevap vermektir. Çalışmamızda, HeLa hücrelerinde MTT ve Akridin oranj/propidyum iyodür boyama ile Dox ve PL'nin hücre canlılığı, Real-Time PCR ve Western-Blot ile PTEN (Fostataz and tensin homolog 10) gen/proteininin ekspresyonu üzerindeki etkileri değerlendirilmiştir. Dox ile kombine edilen PL'nin hücre ölümünü arttırmış ve proliferasyonunu baskılamıştır. Tüm deney gruplarında PTEN gen ekspresyonu azalmıştır ancak PL ve Dox/PL kombine uygulanan kültürlerde PTEN protein fosforilasyonu artmıştır. PL uygulamasının bir tümör baskılayıcı olan PTEN aktivasyonunu arttırması tedavide Dox etkinliğini arttırmak amacı ile kullanılabileceğini göstermektedir.

Anahtar Kelimeler

Doksorubisin, Piperlongumin, PTEN

Proje Numarası

NKUBAP.01.YL.20.267

Kaynakça

• Alves, A. C., Magarkar, A., Horta, M., Lima, J., Bunker, A., Nunes, C., & Reis, S. (2017). Influence of doxorubicin on model cell membrane properties: insights from in vitro and in silico studies. Scientific Reports, 7(1), 6343. doi:10.1038/s41598-017-06445-z
• Chen, Y. J., Kuo, C. C., Ting, L. L., Lu, L. S., Lu, Y. C., Cheng, A. J., …, & Chiou, J. F. (2018). Piperlongumine inhibits cancer stem cell properties and regulates multiple malignant phenotypes in oral cancer. Oncology Letters, 15(2), 1789-1798. doi:10.3892%2Fol.2017.7486
• Fruman, D. A., Chiu, H., Hopkins, B. D., Bagrodia, S., Cantley, L. C., & Abraham, R. T. (2017). The PI3K pathway in human disease. Cell, 170(4), 605-635. doi:10.1016%2Fj.cell.2017.07.029
• Gong, L. H., Chen, X. X., Wang, H., Jiang, Q. W., Pan, S. S., Qiu, J. G., …, & Yan, X. J. (2014). Piperlongumine induces apoptosis and synergizes with cisplatin or paclitaxel in human ovarian cancer cells. Oxidative Medicine and Cellular Longevity, 2014, 906804. doi:10.1155/2014/906804
• Guamán-Ortiz, L. M., Orellana, M. I. R., & Ratovitski, E. A. (2017). Natural compounds as modulators of non-apoptotic cell death in cancer cells. Current Genomics, 18(2), 132-155. doi:10.2174/1389202917666160803150639
• Karki, K., Hedrick, E., Kasiappan, R., Jin, U. H., & Safe, S. (2017). Piperlongumine induces reactive oxygen species (ROS)-dependent downregulation of specificity protein transcription factors. Cancer Prevention Research (Phila), 10(8), 467-477. doi:10.1158/1940-6207.CAPR-17-0053
• Kumar, S., & Agnihotri, N. (2019). Piperlongumine, a piper alkaloid targets Ras/PI3K/Akt/mTOR signaling axis to inhibit tumor cell growth and proliferation in DMH/DSS induced experimental colon cancer. Biomedicine & Pharmacotherapy, 109, 1462-1477. doi:10.1016/j.biopha.2018.10.182
• Momparler, R. L., Karon, M., Siegel, S. E., & Avila, F. (1976). Effect of adriamycin on DNA, RNA, and protein synthesis in cell-free systems and intact cells. Cancer Research, 36(8), 2891-2895.
• Moon, S. K., Kim, H. M., & Kim, C. H. (2004). PTEN induces G1 cell cycle arrest and inhibits MMP-9 expression via the regulation of NF-kappaB and AP-1 in vascular smooth muscle cells. Archives of Biochemistry and Biophysics, 421(2), 267-276. doi:10.1016/j.abb.2003.11.007
• Myers, M. P., & Tonks, N. K. (1997). PTEN: sometimes taking it off can be better than putting it on. The American Journal of Human Genetics, 61(6), 1234-1238. doi:10.1086/301659
• Nakayama, M., Inoue, T., Naito, M., Nakayama, K., & Ohara, N. (2015). Attenuation of the phosphatidylinositol 3-kinase/Akt signaling pathway by Porphyromonas gingivalis gingipains RgpA, RgpB, and Kgp. Journal of Biological Chemistry, 290(8), 5190-5202. doi:10.1074/jbc.M114.591610
• Niu, M., Xu, X., Shen, Y., Yao, Y., Qiao, J., Zhu, F., …, & Xu, K. (2015). Piperlongumine is a novel nuclear export inhibitor with potent anticancer activity. Chemico Biological Interactions, 237, 66-72. doi:10.1016/j.cbi.2015.05.016
• Piska, K., Gunia-Krzyżak, A., Koczurkiewicz, P., Wójcik-Pszczoła, K., & Pękala, E. (2018). Piperlongumine (piplartine) as a lead compound for anticancer agents - Synthesis and properties of analogues: A mini-review. European Journal of Medicinal Chemistry, 156, 13-20. doi:10.1016/j.ejmech.2018.06.057
• Raj, L., Ide, T., Gurkar, A. U., Foley, M., Schenone, M., Li, X., …, & Lee, S. W. (2011). Selective killing of cancer cells by a small molecule targeting the stress response to ROS. Nature, 475(7355), 231–234. (Retraction published Nature. 2018 Sep;561(7723):420) doi:10.1038/nature10167
• Seber, S., Sirin, D. Y., Yetisyigit, T., & Bilgen, T. (2020). Piperlongumine increases the apoptotic effect of doxorubicin and paclitaxel in a cervical cancer cell line. Nigerian Journal of Clinical Practice, 23(3), 386-391. doi:10.4103/njcp.njcp_80_19
• Stambolic, V., Suzuki, A., de la Pompa, J. L., Brothers, G. M., Mirtsos, C., Sasaki, T., …, & Mak, T. W. (1998). Negative regulation of PKB/Akt-dependent cell survival by the tumor suppressor PTEN. Cell, 95(1), 29-39. doi:10.1016/S0092-8674(00)81780-8
• Steel, R. G. D. & Torrie, J. H. (1960). Principles and Procedures of Statistics, With Special References to the Biological Sciences. New York, USA: McGRAW-HILL Book Company.
• Tamura, M., Gu, J., Matsumoto, K., Aota, S., Parsons, R., & Yamada, K. M. (1998). Inhibition of cell migration, spreading, and focal adhesions by tumor suppressor PTEN. Science, 280(5369), 1614-1617. doi:10.1126/science.280.5369.1614
• Takemura, G., & Fujiwara, H. (2007). Doxorubicin-induced cardiomyopathy from the cardiotoxic mechanisms to management. Progress in Cardiovascular Diseases, 49(5), 330-352. doi:10.1016/j.pcad.2006.10.002
• Wang, H., Jiang, H., Corbet, C., De Mey, S., Law, K., Gevaert, T., Feron, O., & De Ridder, M. (2019). Piperlongumine increases sensitivity of colorectal cancer cells to radiation: Involvement of ROS production via dual inhibition of glutathione and thioredoxin systems. Cancer Letters, 450, 42-52. doi:10.1016/j.canlet.2019.02.034
• Xia, Y., Xu, T., Zhao, M., Hua, L., Chen, Y., Wang, C., Tang, Y., & Zhu, B. (2018). Delivery of doxorubicin for human cervical carcinoma targeting therapy by folic acid-modified selenium nanoparticles. International Journal of Molecular Sciences, 19(11), 3582. doi:10.3390/ijms19113582
• Zhou, W., Fu, X. Q., Zhang, L. L., Zhang, J., Huang, X., Lu, H. U., …, & Yu, H. G. (2013). The AKT1/NF-kappaB/Notch1/PTEN axis has an important role in chemoresistance of gastric cancer cells. Cell Death & Disease, 4, e847. doi:10.1038/cddis.2013.375