Laboratório de Bioquímica Celular, Universidade Federal de São João del Rei, Campus Centro-Oeste, Divinópolis, MG, Brazil; Laboratório de Membranas e ATPases, Universidade Federal de São João del Rei, Campus Centro-Oeste, Divinópolis, MG, Brazil.
Laboratório de Bioquímica Celular, Universidade Federal de São João del Rei, Campus Centro-Oeste, Divinópolis, MG, Brazil.
Steroids. 2019 Dec;152:108490. doi: 10.1016/j.steroids.2019.108490. Epub 2019 Sep 6.
Bufadienolide compounds have been used for growth inhibition and apoptosis induction in tumor cells. Those families of cardiotonic steroids can bind the Na,K-ATPase, causing its inhibition. The use of bufadienolides is widely described in the literature as an anticancer function. The aim of this study was to evaluate the effects of bufadienolides and alkaloid isolated from venom samples from R. marina on tumor cells. We performed cytotoxicity assay in MDA-MB-231 and TOV-21G cells and evaluated the activity of Caspases (3 and 9), Na, K-ATPase, PMCA and SERCA. Four compounds were extrated from the venom of R. marina. The compound 1 showed higher cytotoxicity in MDA-MB-231cells. Compound 1 also showed activation of Caspase 3 and 9. This compound caused an inhibition of the activity and expression of Na, K-ATPase, and also showed activation of both caspase-9 and caspase-3 in MDA-MB-231 cells. We also observed that Compound 1 had a direct effect on some ATPases, such as Na, K-ATPase, PMCA and SERCA. Compound 1 was able to inhibit the activity of the purified Na, K-ATPase enzyme from the concentration of 5 µM. It also caused inhibition of PMCA at all concentrations tested (1 nM-30 µM). However, the compound 1 led to an increase of the activity of purified SERCA between the concentrations of 7.5-30 µM. Thus, we present a Na, K-ATPase and PMCA inhibitor, which may lead to the activation of caspases 3 and 9, causing the cells to enter into apoptosis. Our study suggests that compound 1 may be an interesting molecule as an anticancer agent.
蟾毒配基化合物已被用于抑制肿瘤细胞生长和诱导细胞凋亡。这类强心甾体可以与 Na，K-ATP 酶结合，导致其抑制。蟾毒配基的使用在文献中被广泛描述为抗癌功能。本研究旨在评估从 R. marina 毒液样本中分离的蟾毒配基和生物碱对肿瘤细胞的影响。我们在 MDA-MB-231 和 TOV-21G 细胞中进行了细胞毒性测定，并评估了 Caspase（3 和 9）、Na，K-ATP 酶、PMCA 和 SERCA 的活性。从 R. marina 毒液中提取了 4 种化合物。化合物 1 在 MDA-MB-231 细胞中表现出更高的细胞毒性。化合物 1 还显示 Caspase 3 和 9 的激活。该化合物导致 Na，K-ATP 酶的活性和表达受到抑制，同时在 MDA-MB-231 细胞中还激活了 caspase-9 和 caspase-3。我们还观察到化合物 1 对一些 ATP 酶（如 Na，K-ATP 酶、PMCA 和 SERCA）具有直接作用。化合物 1 能够从 5 µM 浓度抑制纯化的 Na，K-ATP 酶的活性。它还在所有测试浓度（1 nM-30 µM）下抑制 PMCA。然而，化合物 1 在 7.5-30 µM 浓度之间导致纯化 SERCA 的活性增加。因此，我们提出了一种 Na，K-ATP 酶和 PMCA 抑制剂，它可能导致 caspase 3 和 9 的激活，导致细胞进入凋亡。我们的研究表明，化合物 1 可能是一种有前途的抗癌药物。
