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

Abstract

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.

Betulinic acid may modulate autophagy in renal cell carcinoma cells

“…According to the results of our previous study (22), betulinic acid dramatically decreased cell viability in ACHN and CAKI-2 cells. However, betulinic acid did not have a signifi cant effect on cell viability of healthy cells.…”

“…In our study conducted in our laboratory in previous years, we determined the cytotoxic effect of betulinic acid on renal cell carcinoma cells by WST-1 assay (22). Cells were seeded in 96-well plate at 1x10 4 cells per well and incubated for 24 h. Following the end of the incubation period, cells were treated with a progressive dosage (1-50 μM) of betulinic acid and incubated for 24 h, 48 h and 72 h. After each incubation period, 10 μl WST-1 solution was added in each well.…”

The inhibitory effect of betulinic acid on epithelial–mesenchymal transition pathway in renal cell carcinoma

“…The expressions of these markers are majorly controlled by one of the SNAIL proteins; SNAIL-1, which binds to E-box DNA sequences through carboxy terminal zing-nger domains of the genes [22]. On the other hand, Syndecan-2 (SDC-2) is a cell surface heparan sulfate proteoglycan, which has also been determined to have a role in differentiation of tumor cells during EMT and induce invasive phenotype in various studies, apart from its role in tissue and organ differentiation and embryonic angiogenesis [23,24]. Although there are many studies in the literature on the expression of SDC-2 in different cancer types, the relationship between SDC-2 and renal cancer has not yet been demonstrated.…”