Arzu ERGEN., ELİF SİNEM İPLİK, BARIŞ ERTUĞRUL, Merve Nur ATAS, Goksu KASARCI, BEDİA ÇAKMAKOĞLU

Examination of the apoptotic effects of betulinic acid on renal cancer cell lines

Objective: Renal cancers are highly resistant to standard hormonal therapy, radiotherapy, and chemotherapy, and the survival rates are extremely low. Betulinic acid is a pentasilic triterpenoid saponin of lupine type obtained from various natural plants, especially from the shell of Betula plant. Betulinic acid was shown both in in vivo, and in vitro to have the ability to induce apoptotic pathways causing no toxicity for normal cells, and also has immunomodulatory effects. The aim of the present project is to investigate the anticancer effects of betulinic acid on CAKI-2 (ATCC® HTB-47™; clear cell renal carcinoma), ACHN (ATCC® CRL-1611™; renal cell adenocarcinoma) and MRC-5 (ATCC® CCL-171™: normal lung fibroblast) cell lines. Materials and Methods: The dose, and time-dependent cell viability was determined using the WST-1 test first in cell lines, and then apoptotic activity was determined with Annexin-V, apoptosis related nucleosomal enrichment factor levels, and Caspase 3 / BCA activity. Results: Betulinic acid reduced the CAKI-2, and ACHN cell viability in dose, and time-dependent manner inducing the apoptotic pathway. Conclusion: Researchers in the present study concluded in accordance with the results of Annexin-V, apoptosis-associated nucleosomal enrichment factor levels and Caspase 3 / BCA activity that betulinic acid triggered the apoptosis in both renal cancer cell lines, especially by the Caspase 3 activity.

Keywords:

Renal cancer, Betulinic acid, Apoptosis, ACHN, CAKI-2,

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Phillips DR, Rasbery JM, Bartel B, Matsuda SP. Biosynthetic diversity in plant triterpene cyclization. Curr Opin Plant Biol 2006; 9:305-14. doi: 10.1016/j.pbi.2006.03.004
Cmoch P, Pakulski Z, Swaczynova J, Strnad M. Synthesis of lupane-type saponins bearing mannosyl and 3,6-branched trimannosyl residues and their evaluation as anticancer agents. Carbohydr Res 2008; 343: 995-1003. doi: 10.1016/j. carres.2008.02.011.
Zhang X, Hu J, Chen Y. Betulinic acid and the pharmacological effect of tumor supressions. Mol Med Rep 2016; 14:4489 – 95. doi: 10.3892/mmr.2016.5792.
Fulda S, Scaffidi C, Susin SA, et al. Activation of mitochondria and release of mitochondrial apoptogenic factors by betulinic acid. J Biol Chem 1998; 273:33942-48. doi: 10.1074/ jbc.273.51.33942
Fulda S, Galluzzi L, Kroemer G. Targeting mitochondria for cancer therapy. Nat Rev Drug Discov 2010; 9:447-64. doi: 10.1038/nrd3137.
Fulda S. Targeting apoptosis for anticancer therapy. Semin Cancer Biol 2015; 31:84-88. doi: 10.1016/j. semcancer.2014.05.002.
Hsu TI, Wang MC, Chen SY, et al. Betulinic acid decreases specificity protein 1 (Sp1) level via increasing the sumoylation of Sp1 to inhibit lung cancer growth. Mol Pharmacol 2012; 82:1115-28. doi: 10.1124/mol.112.078485.
Struh CM, Jager S, Schempp CM, Scheffler A, Martin SF. A novel triterpene extract from mistletoe induces rapid apoptosis in murine B16.F10 melanoma cells. Phytother Res 2012; 26:1507-12. doi: 10.1002/ptr.4604.
Soica C, Danciu C, Savoiu-Balint G, et al. Betulinic acid in complex with a gamma-cyclodextrin derivative decreases proliferation and in vivo tumor development of nonmetastatic and metastatic B164A5 cells. Int J Mol Sci 2014; 15:8235-55. doi: 10.3390/ijms15058235.
Wang P, Li Q, Li K, et al. Betulinic acid exerts immunoregulation and anti-tumor effect on cervical carcinoma (U14) tumorbearing mice. Pharmazie 2012; 67:733-9. doi.org/10.1691/ ph.2012.1822
Kowalczyk T, Sitarek P, Skała E, et al. Induction of apoptosis by in vitro and in vivo plant extracts derived from Menyanthes trifoliata L. in human cancer cells. Cytotechnology 2019; 71:165-80. doi: 10.1007/s10616.018.0274-9.
Zhao Z, Wang J, Tang J, et al. JNK and Akt-mediated Puma expression in the apoptosis of cisplatin-resistant ovarian cancer cells. Biochem J. 2012; 444: 291-301. doi: 10.1042/ BJ20111855.
Li L, Du Y, Kong X, et al. Lamin B1 is a novel therapeutic target of betulinic acid in pancreatic cancer. Clin Cancer Res 2013; 1:4651-61. doi: 10.1158/1078-0432.CCR-12-3630.
Saeed MEM, Mahmoud N, Sugimoto Y, Efferth T, Abdel- Aziz H. Betulinic Acid Exerts Cytotoxic Activity Against Multidrug-Resistant Tumor Cells via Targeting Autocrine Motility Factor Receptor (AMFR). Front Pharmacol. 2018; 9:481. doi: 10.3389/fphar.2018.00481.
Zeng A, Hua H, Liu L, Zhao J. Betulinic acid induces apoptosis and inhibits metastasis of human colorectal cancer cells in vitro and in vivo. Bioorg Med Chem 2019;27:2546-52. doi: 10.1016/j.bmc.2019.03.033.
Yang C, Li Y, Fu L, Jiang T, Meng F. Betulinic acid induces apoptosis and inhibits metastasis of human renal carcinoma cells in vitro and in vivo. J Cell Biochem 2018;119:8611-22. doi: 10.1002/jcb.27116.
Wang R, Yang M, Li G, et al. Paclitaxel-betulinic acid hybrid nanosuspensions for enhanced anti-breast cancer activity. Colloids Surf B Biointerfaces 2019;174:270-9. doi: 10.1016/j. colsurfb.2018.11.029.
Gao Y, Ma Q, Ma YB, et al. Betulinic acid induces apoptosis and ultrastructural changes in MDA-MB-231 breast cancer cells. Ultrastruct Pathol 2018;42:49-54. doi: 10.1080/01913.123.2017.1383548.