Proanthocyanidins,Novel bioactive components for prevention of melanoma invasion

原花青素，预防黑色素瘤侵袭的新型生物活性成分

• 批准号: 8442533
• 负责人: SANTOSH KUMAR KATIYAR
• 金额: $ 19.13万
• 依托单位: UNIVERSITY OF ALABAMA AT BIRMINGHAM
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-01-02 至 2014-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8442533
• 关键词: Affect; Animal Model; BRAF gene; Biological Assay; Cause of Death; Cell Culture Techniques; Cell Line; Cell Survival; Cells; Cessation of life; Chemopreventive Agent; Child; Clinical; Consumption; Data; Development; Diagnosis; Diet; Dietary Component; Disease; Dose; Effectiveness; Exhibits; Future; Goals; Growth; Human; In Vitro; Incidence; Individual; Knowledge; Light; Malignant - descriptor; Malignant Neoplasms; Mediating; Melanoma Cell; Metastatic Melanoma; Modeling; Molecular; Mus; Mutate; Mutation; Neoplasm Metastasis; Nuclear; Nude Mice; Oncogenes; Outcome Study; Patients; Pharmaceutical Preparations; Prevention; Primary Neoplasm; Proanthocyanidins; Reporting; Research Design; Resistance; Safety; Sampling; Signal Pathway; Signal Transduction; Skin; Skin Cancer; Testing; Toxic effect; Trypan Blue; base; carcinogenesis; cell motility; grape seed; high risk; in vivo; in vivo Model; innovation; interest; killings; melanocyte; melanoma; migration; mortality; mouse model; mutant; neoplastic cell; novel; public health relevance; skin disorder; tumor growth

DESCRIPTION (provided by applicant): Melanoma is the leading cause of death from skin disease due, in large part, to its propensity to metastasize. Since metastasis of melanoma is the leading cause of death due to skin diseases, an approach that decreases its invasiveness or metastatic ability may facilitate the development of an effective strategy for its treatment and/or prevention. Some drugs has demonstrated potential in treatment of melanoma but their use as chemopreventive agents is questionable due to long-term safety, resistance and toxicity concerns. Dietary bioactive components that have low toxicity are of interest as chemopreventives, including the proanthocyanidins, which do not exhibit gross toxicity in mice. Proanthocyanidins, which are highly enriched in grape seeds, have anti-skin carcinogenic effects in mouse models, but their effects on metastasis have not been explored. We have found that treatment of some metastatic human melanoma cells (A375, Hs294t) with grape seed proanthocyanidins (GSPs) inhibited their migration, as determined using a Boyden chamber assay, suggesting the possibility that GSPs may be effective in inhibiting metastasis. As mutations in b-catenin are one of the most common alterations associated with melanoma development and metastasis and mutated b- catenin is associated with aggressive tumor growth, we examined the effects of the b-catenin status and found that the GSPs-mediated inhibition of melanoma cell invasion was associated with inactivation of b-catenin and that melanoma cells that have activated b-catenin (Mel 1241) showed significantly greater invasion than those melanoma cells that have inactivated b-catenin (Mel 1011). Based on our preliminary data, we have formulated the innovative hypothesis that the inhibition of migration or invasiveness of melanoma cells by GSPs is mediated, at least in part, through inactivation of b-catenin and that is critical for their chemopreventive effects on melanoma metastasis. Our long-term goal is to fully test this hypothesis. The objectives of this exploratory R21 application are o verify and extend our preliminary results using in vitro cell culture and in vivo athymic nude mouse model in order to generate the data necessary for the design of studies that will rigorously test the central hypothesis in mouse model and establish the molecular basis for the observed data. We propose two complementary Specific Aims: (1) To determine whether GSPs inhibit cell invasion of human melanoma cell lines with different mutations, and whether inhibition of cell invasion by GSPs requires inactivation of b-catenin and its signaling. Normal human epidermal melanocytes will serve as a control, and (2) To determine the effect of dietary GSPs on the invasive potential of human melanoma cells in vivo in athymic nude mice, and to ascertain whether the inhibitory effect of GSPs on cell metastasis is associated with inactivation of b-catenin in this model. RELEVANCE: These studies will generate knowledge concerning the effects of GSPs on melanoma invasiveness/metastasis in vivo and the mechanism of action that is necessary for future studies of their potential long-term use for suppression of metastatic melanoma.

描述（由申请人提供）：黑色素瘤是皮肤病死亡的主要原因，很大程度上是由于其转移倾向。由于黑色素瘤的转移是皮肤病导致死亡的主要原因，因此降低其侵袭性或转移能力的方法可能有助于制定有效的治疗策略和/或 预防。一些药物已被证明具有治疗黑色素瘤的潜力，但由于长期安全性、耐药性和毒性问题，它们作为化学预防剂的使用值得怀疑。低毒性膳食生物活性成分作为化学预防剂受到关注，其中包括原花青素，它在小鼠中不表现出明显的毒性。葡萄籽中高度丰富的原花青素在小鼠模型中具有抗皮肤致癌作用，但其对转移的影响尚未被探索。我们发现，用博伊登室试验测定，用葡萄籽原花青素 (GSP) 处理一些转移性人类黑色素瘤细胞（A375、Hs294t）可抑制其迁移，这表明 GSP 可能有效抑制转移。由于β-连环蛋白突变是与黑色素瘤发展和转移相关的最常见的改变之一，并且突变的β-连环蛋白与侵袭性肿瘤生长相关，因此我们检查了b-连环蛋白状态的影响，发现GSP介导的黑色素瘤细胞侵袭抑制与b-连环蛋白失活相关，并且激活了b-连环蛋白（Mel 1241）的黑色素瘤细胞表现出显着更大的侵袭性 比那些使 b-连环蛋白失活的黑色素瘤细胞 (Mel 1011) 更有效。根据我们的初步数据，我们提出了创新假设，即 GSP 对黑色素瘤细胞迁移或侵袭的抑制至少部分是通过 β-连环蛋白失活介导的，这对于它们对黑色素瘤转移的化学预防作用至关重要。我们的长期目标是充分检验这一假设。这一探索性 R21 应用的目的是使用体外细胞培养和体内无胸腺裸鼠模型验证和扩展我们的初步结果，以便生成设计研究所需的数据，这些数据将严格测试小鼠模型的中心假设并为观察到的数据建立分子基础。我们提出了两个互补的具体目标：（1）确定GSP是否抑制具有不同突变的人黑色素瘤细胞系的细胞侵袭，以及GSP抑制细胞侵袭是否需要b-连环蛋白及其信号传导失活。以正常人表皮黑素细胞作为对照，(2)在无胸腺裸鼠体内测定膳食GSPs对人黑色素瘤细胞体内侵袭能力的影响，并确定GSPs对细胞转移的抑制作用是否与该模型中b-catenin的失活有关。相关性：这些研究将产生有关 GSP 对体内黑色素瘤侵袭/转移的影响以及作用机制的知识，这对于未来长期研究其抑制转移性黑色素瘤的潜在作用是必要的。