Spatane diterpinoid from the brown algae, Stoechospermum marginatum induces apoptosis via ROS induced mitochondrial mediated caspase dependent pathway in murine B16F10 melanoma cells
Spatane diterpinoid from the brown algae, Stoechospermum marginatum induces apoptosis via ROS induced mitochondrial mediated caspase dependent pathway in murine B16F10 melanoma cells
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DOI：
10.1002/mc.22463
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发表时间：
2016-12
影响因子：
4.6
通讯作者：
L. Velatooru;Chinnababu Baggu;Venkateswara Rao Janapala
中科院分区：
文献类型：
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作者：
L. Velatooru;Chinnababu Baggu;Venkateswara Rao Janapala
已知从褐藻Stoechospermum marginatum中分离的Spatane diterpinoids对人类癌细胞系和小鼠黑色素瘤细胞具有细胞毒性作用;到目前为止，diterpinoids的潜在凋亡机制仍然不清楚。因此，在本研究中，主要在B16 F10黑色素瘤细胞中研究了spatane diterpinoid，5（R），19-diacetoxy-15，18（R和S），dihydro spata-13，16（E）-diene（DDSD）的凋亡机制，因为它们比THP 1，U937，COLO 205和HL 60细胞更容易受到DDSD的影响。用DDSD处理B6 F10细胞导致形态学改变、核浓缩和DNA片段化，从而以浓度依赖性方式抑制细胞生长。数据表明，DDSD诱导ROS的产生，从而导致Bax/Bcl-2比值的改变，破坏线粒体内跨膜电位（Δ Km），导致细胞色素c重新分布到细胞质，并激活caspase介导的凋亡途径。流式细胞术分析清楚地表明DDSD诱导磷脂酰丝氨酸外化并介导细胞周期中的“S期”停滞。DDSD通过下调PI 3 K/AKT信号通路诱导细胞凋亡。在携带B16 F10黑色素瘤的C57 BL/6小鼠中评价DDSD的抗肿瘤活性。它以剂量依赖性方式有效抑制肿瘤生长（体积和重量），但没有明显的毒性作用。分别通过显微镜和TUNEL法评估治疗小鼠中肿瘤组织的形态学和凋亡状态。我们的研究表明DDSD治疗恶性黑色素瘤的治疗潜力和抗癌药物的新来源。© 2016 Wiley Periodicals，Inc.
Spatane diterpinoids isolated from the brown marine algae Stoechospermum marginatum were known to have cytotoxic effects in human cancerous cell lines and murine melanoma cells; the underling apoptotic mechanism of diterpinoids still remains unclear so far. Thus, in the present study, the apoptotic mechanism of a spatane diterpinoid, 5(R), 19‐diacetoxy‐15,18(R and S), dihydro spata‐13, 16(E)‐diene (DDSD) was investigated mainly in B16F10 melanoma cells because they were most susceptible to DDSD than THP1, U937, COLO205, and HL60 cells. The treatment of B6F10 cells with DDSD resulted in morphological alterations, nuclear condensation, and DNA fragmentation, which leads to cell growth inhibition in a concentration‐dependent manner. Data indicate that DDSD induced the generation of ROS, consequentially caused alteration in Bax/Bcl‐2 ratio that disrupted the inner mitochondrial transmembrane potential (ΔΨm) resulting in cytochrome c redistribution to the cytoplasm and activation of caspase‐mediated apoptotic pathway. Flow cytometric analysis clearly indicated that the DDSD inducing phosphatidylserine externalization and mediated “S‐phase” arrest in cell cycle. In addition, results also found that DDSD induced apoptosis through deregulating PI3K/AKT signaling pathway. The anti‐tumor activity of DDSD was evaluated in C57BL/6 mice bearing B16F10 melanoma. It effectively inhibited tumor growth (volume and weight) in a dose dependent manner, yet without apparent toxic effects. Morphology and apoptotic status of tumor tissues in the treated mice were assessed by microscopy and TUNEL assay, respectively. Our study shows a therapeutic potential of DDSD for the treatment of malignant melanoma and a new source of anticancer drugs. © 2016 Wiley Periodicals, Inc.