The effects of sertraline and sorafenib on P-glycoprotein expression and rhodamine 123 accumulation in liver cancer cells

Yıl 2022, Cilt: 15 Sayı: 1, 56 - 65, 30.04.2022

Yaprak Dönmez Çakıl , İlayda Altun , Elif Tekin İşlerel , Zeynep Ozunal

https://doi.org/10.26559/mersinsbd.1001821

Öz

Aim: Hepatocellular carcinoma (HCC) is among the most common lethal cancer type with an increasing incidence all over the world. In the majority of cases, advanced stage diagnosis and drug resistance are the main issues that limit the survival rate. P-glycoprotein (P-gp), an ATP-dependent efflux pump, is associated with numerous drug resistance patterns in cancer. Rhodamine 123 (rh123) is a fluorescent dye used as a reference substrate for measurement of P-gp activity in various studies. Sorafenib is the first approved systemic therapy for HCC treatment. Novel combined approaches of sorafenib with cytotoxic drugs are extensively studied to increase its effectiveness and overcome drug resistance. Recently, the synergistic effects of sorafenib and antidepressant sertraline on HepG2 cell proliferation and death were shown. Method: In the present study, we investigated the possible effects of sorafenib and sertraline as single agents or in combination on P-gp expression and rh123 accumulation/efflux by qPCR and flow cytometry, respectively. Results: Sorafenib administration significantly decreased both P-gp gene expression and rh123 accumulation in cells, while sertraline increased P-gp mRNA levels, it did not cause a significant change in rh123 accumulation. Co-administration of the two drugs did not cause any alterations in P-gp gene expression and rh123 accumulation compared to the control group. Conclusion: The results suggested that the presence of other mechanisms rather than P-gp associated effects are responsible for the synergistic activity of sorafenib and sertraline.

Anahtar Kelimeler

Sertraline , sorafenib , synergy , p-glycoprotein , HepG2 , rhodamine123

Proje Numarası

1919B012001071

Sertralin ve sorafenibin karaciğer kanseri hücrelerinde P-glikoprotein gen ekspresyonuna ve rodamin 123 birikimine etkileri

Öz

Amaç: Hepatoselüler karsinom (HCC), tüm dünyada görülme sıklığı giderek artan, en yaygın ölümcül kanser türleri arasında yer almaktadır. Vakaların büyük çoğunluğunda ileri evrede tanı ve ilaç direnci hayatta kalma oranını sınırlayan temel sorunlardır. ATP bağımlı bir efluks (dışa atım) pompası olan P-glikoprotein (P-gp), kanserde çok sayıda ilaç direnci modeli ile ilişkilidir. Rodamin 123 (rh123) floresan bir boyadır ve bir referans P-gp substratı olarak birçok araştırmada P-gp aktivitesini incelemek amacıyla kullanılmaktadır. Sorafenib, HCC tedavisi için onaylanmış ilk sistemik tedavidir. Etkinliğini artırmak ve ilaç direncini azaltmak için farklı ilaçlarla beraber kullanımları araştırılmaktadır. Bu kapsamda, daha önce sorafenib ve antidepresan sertralinin HepG2 hücre proliferasyonu ve ölümü üzerinde sinerjistik etkileri gösterilmiştir. Yöntem: Bu çalışmada, HepG2 hücrelerinde, sorafenib ve sertralinin tek başına veya birlikte uygulanması sonrası, qPCR ve akış sitometrisi ile P-gp gen ekspresyonu ve rh123 birikimi/efluks araştırılmıştır. Bulgular: Sorafenib uygulaması hem P-gp gen ekspresyonu hem de hücrelerde rh123 birikimini anlamlı olarak azaltırken, sertralin tek başına kullanıldığında P-gp mRNA seviyelerini arttırmış, rh123 birikiminde ise anlamlı bir değişikliğe yol açmamıştır. İki ilacın birlikte uygulanması ise, kontrol grubuna kıyasla P-gp gen ekspresyonunda ve rh123 birikiminde herhangi bir değişikliğe neden olmamıştır. Sonuç: Sonuçlar, sorafenib ve sertralinin antiproliferatif sinerjistik etkilerinde P-gp ile ilişkili etkenlerden ziyade farklı mekanizmaların rol alabileceğini göstermiştir.

Anahtar Kelimeler

Sertraline , sorafenib , sinerjist etki , P-glikoprotein , HepG2 , rodamin 123

Destekleyen Kurum

TÜBİTAK 2209-A Üniversite Öğrencileri Araştırma Projeleri Destekleme Programı

Proje Numarası

1919B012001071

Teşekkür

İstatistik analizlerinde değerli katkılarından dolayı Dr. Pınar Bıçaksız’a teşekkür ederiz.

Kaynakça

• Llovet JM, Kelley RK, Villanueva A, et al. Hepatocellular carcinoma. Nat Rev Dis Primers. 2021;7(1):6. doi:10.1038/s41572-020-00240-3
• Tang W, Chen Z, Zhang W, et al. The mechanisms of sorafenib resistance in hepatocellular carcinoma: theoretical basis and therapeutic aspects. Signal Transduct Target Ther. 2020;5(1):87. doi:10.1038/s41392-020-0187-x
• Luo XY, Wu KM, He XX. Advances in drug development for hepatocellular carcinoma: clinical trials and potential therapeutic targets. J Exp Clin Cancer Res. 2021;40(1):172. doi:10.1186/s13046-021-01968-w
• Lang L. FDA approves sorafenib for patients with inoperable liver cancer. Gastroenterology. 2008;134(2):379. doi:10.1053/j.gastro.2007.12.037
• Cabral LKD, Tiribelli C, Sukowati CHC. Sorafenib Resistance in Hepatocellular Carcinoma: The Relevance of Genetic Heterogeneity. Cancers (Basel). 2020;12(6). doi:10.3390/cancers12061576
• Wang Z, Zhao Z, Wu T, Song L, Zhang Y. Sorafenib-irinotecan sequential therapy augmented the anti-tumor efficacy of monotherapy in hepatocellular carcinoma cells HepG2. Neoplasma. 2015;62(2):172-9. doi:10.4149/neo_2015_022
• Abou-Alfa GK, Johnson P, Knox JJ, et al. Doxorubicin plus sorafenib vs doxorubicin alone in patients with advanced hepatocellular carcinoma: a randomized trial. Jama. 2010;304(19):2154-60. doi:10.1001/jama.2010.1672
• Liu Y, Yue H, Xu S, et al. First-line gemcitabine and oxaliplatin (GEMOX) plus sorafenib, followed by sorafenib as maintenance therapy, for patients with advanced hepatocellular carcinoma: a preliminary study. Int J Clin Oncol. 2015;20(5):952-9. doi:10.1007/s10147-015-0796-5
• Seelig A. P-Glycoprotein: One Mechanism, Many Tasks and the Consequences for Pharmacotherapy of Cancers. Front Oncol. 2020;10:576559. doi:10.3389/fonc.2020.576559
• Zhou SF. Structure, function and regulation of P-glycoprotein and its clinical relevance in drug disposition. Xenobiotica. 2008;38(7-8):802-32. doi:10.1080/00498250701867889
• Jouan E, Le Vee M, Denizot C, Da Violante G, Fardel O. The mitochondrial fluorescent dye rhodamine 123 is a high-affinity substrate for organic cation transporters (OCTs) 1 and 2. Fundam Clin Pharmacol. 2014;28(1):65-77. doi:10.1111/j.1472-8206.2012.01071.x
• Jouan E, Le Vée M, Mayati A, Denizot C, Parmentier Y, Fardel O. Evaluation of P-Glycoprotein Inhibitory Potential Using a Rhodamine 123 Accumulation Assay. Pharmaceutics. 2016;8(2):12.
• Geeraerts SL, Kampen KR, Rinaldi G, et al. Repurposing the Antidepressant Sertraline as SHMT Inhibitor to Suppress Serine/Glycine Synthesis-Addicted Breast Tumor Growth. Mol Cancer Ther. 2021;20(1):50-63. doi:10.1158/1535-7163.Mct-20-0480
• Stapel B, Melzer C, von der Ohe J, et al. Effect of SSRI exposure on the proliferation rate and glucose uptake in breast and ovary cancer cell lines. Sci Rep. 2021;11(1):1250. doi:10.1038/s41598-020-80850-9
• Ozunal ZG, Donmez Cakil Y, Isan H, Saglam E, Aktas RG. Sertraline in combination with sorafenib: A promising pharmacotherapy to target both depressive disorders and hepatocellular cancer. BioFut. 2019;70(4):341-348. doi:10.1556/019.70.2019.39
• Dönmez Cakil Y, Khunweeraphong N, Parveen Z, et al. Pore-exposed tyrosine residues of P-glycoprotein are important hydrogen-bonding partners for drugs. Mol Pharmacol. 2014;85(3):420-8. doi:10.1124/mol.113.088526
• Hoffmann K, Franz C, Xiao Z, et al. Sorafenib modulates the gene expression of multi-drug resistance mediating ATP-binding cassette proteins in experimental hepatocellular carcinoma. Anticancer Res. 2010;30(11):4503-8.
• Ye CG, Yeung JH, Huang GL, et al. Increased glutathione and mitogen-activated protein kinase phosphorylation are involved in the induction of doxorubicin resistance in hepatocellular carcinoma cells. Hepatol Res. 2013;43(3):289-99. doi:10.1111/j.1872-034X.2012.01067.x
• Wei L, Huang N, Yang L, et al. [Sorafenib reverses multidrug resistance of hepatoma cells in vitro]. Nan Fang Yi Ke Da Xue Xue Bao. 2009;29(5):1016-9, 1023.
• Smith HR. Depression in cancer patients: Pathogenesis, implications and treatment (Review). Oncol Lett. 2015;9(4):1509-1514. doi:10.3892/ol.2015.2944
• Beretta GL, Cassinelli G, Pennati M, Zuco V, Gatti L. Overcoming ABC transporter-mediated multidrug resistance: The dual role of tyrosine kinase inhibitors as multitargeting agents. Eur J Med Chem. 2017;142:271-289. doi:10.1016/j.ejmech.2017.07.062
• Hu S, Chen Z, Franke R, et al. Interaction of the multikinase inhibitors sorafenib and sunitinib with solute carriers and ATP-binding cassette transporters. Clin Cancer Res. 2009;15(19):6062-9. doi:10.1158/1078-0432.Ccr-09-0048
• Lagas JS, van Waterschoot RAB, Sparidans RW, Wagenaar E, Beijnen JH, Schinkel AH. Breast Cancer Resistance Protein and P-glycoprotein Limit Sorafenib Brain Accumulation. Molecular Cancer Therapeutics. 2010;9(2):319-326. doi:10.1158/1535-7163.Mct-09-0663
• Huang WC, Hsieh YL, Hung CM, et al. BCRP/ABCG2 inhibition sensitizes hepatocellular carcinoma cells to sorafenib. PLoS One. 2013;8(12):e83627. doi:10.1371/journal.pone.0083627
• Tandia M, Mhiri A, Paule B, et al. Correlation between clinical response to sorafenib in hepatocellular carcinoma treatment and polymorphisms of P-glycoprotein (ABCB1) and of breast cancer resistance protein (ABCG2): monocentric study. Cancer Chemother Pharmacol. 2017;79(4):759-766. doi:10.1007/s00280-017-3268-y
• Wang X, Zhang X, Huang X, Li Y, Wu M, Liu J. The drug-drug interaction of sorafenib mediated by P-glycoprotein and CYP3A4. Xenobiotica. 2016;46(7):651-658. doi:10.3109/00498254.2015.1109160