Combination of cisplatin and bromelain exerts synergistic cytotoxic effects against breast cancer cell line MDA-MB-231 in vitro

Abstract

BackgroundBromelain, which is a cysteine endopeptidase commonly found in pineapple stems, has been investigated as a potential anti-cancer agent for the treatment of breast cancer. However, information pertaining to the effects of combining bromelain with existing chemotherapeutic drugs remains scarce. This study aimed to investigate the possible synergistic cytotoxic effects of using bromelain in combination with cisplatin on MDA-MB-231 human breast cancer cells.MethodMDA-MB-231 cells were treated with different concentrations (0.24–9.5 µM) of bromelain or cisplatin alone, as well as four different combinations of these two agents to assess their individual and combination effects after 24 and 48 h. Cell viability was analyzed using an MTT assay. The induction of apoptosis was assessed using cell cycle analysis and an Annexin V-FITC assay. The role of the mitochondrial membrane potential in the apoptotic process was assessed using a JC-1 staining assay. Apoptotic protein levels were assessed by western blot analysis and proteome profiling using an antibody array kit.ResultsSingle-agent treatment with cisplatin or bromelain led to dose- and time-dependent decreases in the viability of the MDA-MB-231 cells at 24 and 48 h. Furthermore, most of the combinations evaluated in this study displayed synergistic effects against MDA-MB-231 cells at 48 h, with combination 1 (bromelain 2 µM + cisplatin 1.5 µM) exhibiting the greatest synergistic effect (P = 0.000). The results of subsequent assays indicated that combination 1 treatment induced apoptosis via mitochondria-mediated pathway. Combination 1 also resulted in significant decreases in the levels of several apoptotic proteins such as Bcl-x and HSP70, compared with bromelain (P = 0.002 and 0.000, respectively) or cisplatin (P = 0.000 and 0.001, respectively) single treatment. Notably, MDA-MB-231 cells treated with combination 1 showed increased levels of the pro-apoptotic proteins Bax compared with those treated with bromelain (P = 0.000) or cisplatin single treatment (P = 0.043).ConclusionBromelain in combination with cisplatin synergistically enhanced the induction of apoptosis in MDA-MB-231 cells.

Arachidonic Acid Attenuates Cell Proliferation, Migration and Viability by a Mechanism Independent on Calcium Entry

“…Additionally, changes in the fluorescence of JC-1 were analyzed in order to determine whether AA might evoke mitochondrial potential depolarization. Thus, MDA-MB-231 cells were grown for 48 h in the presence of AA (8 µM) and, at the indicated times (0, 24, and 48 h), cells were subsequently incubated for an additional 30 min with 2 µM of JC-1 [51]. Finally, JC-1 was excited at a 488 nm wavelength and cell images were acquired at the emission wavelengths of 530 nm (JC-1 monomers, "JC-1 green") or 580 nm (JC-1 aggregates, "JC-1 red") using an inverted fluorescence microscopy and 40× WD objective.…”

Anti-cancer effects of Rhizoma Curcumae against doxorubicin-resistant breast cancer cells

“…Cell viability was performed using MTT assay as described previously [ 33 ]. Briefly, exponentially growing cells were seeded in 96-well plates at a density of 2 × 10 4 /well and allowed to attach overnight.…”

Bromelain and Cancer

“…Bu kombinasyon tedavisi, bromelain ya da sisplatinin tekli tedavisi ile karşılaştırıldığında, proapoptotik protein Bax düzeylerinde artışa neden olduğu rapor edilmiştir. 29 Pillai ve ark. tarafından yürütülen bir çalışmada, malign peritoneal mezotelyoma hücreleri laboratuvar koşullarında 72 saat boyunca bromelain ve sitotoksik ilaçlara (sisplatin veya 5-FU) maruz bırakıldığında, bromelain ilavesi ile sisplatinin sitotoksisitesinde önemli ölçüde artış olur iken; 5-FU sitotoksisitesinde anlamlı bir artış olmamıştır.…”