Enhancement of Academic Research at Southern University Baton Rouge

描述(申请人提供)：生物活性、非类固醇、雄激素和雌激素制剂主要存在于豆制品、豆类和全谷物中，以及药茶和根茎中。Kola Acumate，也被牙买加的Ettu人称为Bizzy坚果，是一种据报道影响各种生物过程的“万能”草药， 其中许多是由激素直接或间接调节的。现有的民族植物学信息表明，科拉针叶可能含有具有雌激素和雄激素特性的生物活性化学物质。坊间报道表明，Bizzy坚果可能在许多医疗目的上有用。鉴于其中一些生物活性的激素依赖性，Bizzy坚果中存在的非类固醇雄激素可能是其药用价值的原因。一种药物作为前列腺癌和其他类型癌症的化学预防药物，其成功的关键取决于该药物诱导肿瘤细胞死亡的能力。 一种组织依赖和受体依赖的方式。以AR依赖的方式诱导前列腺细胞凋亡的自然产生的药物将标志着新一代前列腺特异性化学预防药物的诞生。本研究项目的目的是验证Bizzy乙醚提取物(BIZ-2)以可预测的方式调节前列腺功能的假设。因此，这一修订申请的总体目标保持不变，即证明Kola Acumate(Bizzy Nut)提取物含有能够作为前列腺癌化学预防药物的生物活性化合物。这一应用的最初重点是分离和鉴定Bizzy Nut的Biz-2Fr.3组分中的生物活性化合物，并使用三种前列腺癌细胞模型在体外证明其抗癌/化学预防的潜力。为此，制定了两个具体目标。目的1：分离和鉴定Bizzy坚果提取物中与其在前列腺细胞中的抗癌活性相关的活性化合物。本研究的目的是：(1)通过制备高效液相色谱分离和表征BIZ-2Fr.3的生物活性成分；(2)结合UV-Vis、核磁共振光谱、LC-MS鉴定BIZ-2Fr.3中的化合物结构；(3)证明BIZ-2Fr.3中的生物活性物质具有抗癌作用。目的2：鉴定Biz-2Fr.3诱导前列腺细胞凋亡的信号转导途径，探讨Biz-2Fr.3诱导前列腺细胞凋亡的分子机制。本研究的目的是：(1)通过细胞周期分布和细胞周期蛋白表达谱的改变来确定Biz-2Fr.3抗肿瘤活性的机制。(2)以AR+LNCaP、AR-DU145细胞和RWP-1a正常转化细胞系为研究对象，研究线粒体在BIZ-2Fr.3诱导正常前列腺细胞和前列腺癌细胞凋亡中的作用。我们已经为开展拟议的研究做好了充分的准备，因为我们在实验系统方面的经验(见初步结果)和化学、生物和生物化学方面的独特专家科学家团队，以及我们来自路易斯安那州立大学化学和SUBR化学的合作者和导师，他们参与了该项目。我们已经开发并组装了一套工具、关键试剂和细胞系，以确保这一应用的成功。此外，我们的初步数据有力地支持了建议的研究，这些数据验证了指导建议研究的假设。最后，汇集的专业知识、充足的设施、强有力的合作和机构支持将使我们能够在有利于研究成功的研究环境中进行这项研究。

DESCRIPTION (provided by applicant): Biologically active, non-steroidal, androgenic and estrogenic agents are found primarily in soy products, legumes, and whole grains, as well as in medicinal teas and roots. Kola acuminate, also known as Bizzy nut to the Ettu people of Jamaica, is a "cure-all" herbal medicine that reportedly affects a variety of biological processes, many of which are directly or indirectly modulated by hormones. Available ethnobotanical information suggests that Kola acuminate may contain bioactive chemicals that possess estrogenic and androgenic properties. Anecdotal reports suggest that Bizzy nut may be useful for a number of medical purposes. Given the hormonal dependency of some of these biological activities, it is possible that non-steroidal androgen present in the Bizzy nut is responsible for he medicinal value attributed to it. The key to an agent's success, as a chemo preventive agent in the prostate and other types of cancer, relies on the agent's ability to induce tumor cell death in a tissue-dependent and receptor-dependent manner. Naturally occurring agents that induce apoptosis in prostate cells in an AR-dependent manner would signal a new generation of prostate-specific, chemo preventive agents. The goal of this research project is to test the hypothesis that ether extract of Bizzy (Biz-2) modulates prostate function in a predictable manner. Thus, the overall objective of this revised application remains the same, namely demonstrating that the Kola acuminate (Bizzy Nut) extract contains bioactive compounds capable of functioning as chemo preventive agents against prostate cancer. The initial focus of this application is to isolated and identify the bioactive compounds found in the Biz-2Fr.3 fraction of Bizzy Nut and demonstrate, In vitro, its anti- cancer/chemo preventive potential using three prostate cancer cell models. To this end, two Specific Aims were developed. Aim 1: is to isolate and characterize the bioactive compounds in Bizzy nut extract associated with its anti-cancer properties in prostate cells. The goals of this Aim are to (1) Isolate and characterize the bioactive components of Biz-two by preparative HPLC; (2) elucidate the structure of the compounds in Biz-2Fr.3 using a combination of UV-Vis, NMR spectroscopy, LC-MS; and (3) Demonstrate the anti-cancer capability of the bioactive substances in Biz-2Fr.3. Aim 2: is to Identify the Biz-2Fr.3-induced apoptotic signaling pathways and determine the molecular mechanisms by which Biz-2FR.3 induces apoptosis in prostate cells. The goals of this Aim are to; (1) determine the mechanism of Biz-2Fr.3 anti-tumor activity using change in cell cycle distribution and protein expression profiling of cyclines. (2) Determine the involvement of mitochondria in Biz-2Fr.3 induction of apoptosis in normal and cancerous prostate cells using three prostate cell lines, the AR+ LNCaP, the AR- DU145 cells and RWP-1a normal, transformed cell line. We are well prepared to undertake the proposed research because of our experience with the experimental systems (see preliminary results) and the unique team of expert scientists in chemistry, biology and biochemistry as well as our collaborators and mentors from the LSU Chemistry and SUBR chemistry who are involved with the project. We have developed and assembled a set of tools, critical reagents and cell lines that will ensure success of this application. In addition, the proposed study is strongly supported by our preliminary data that validate the hypotheses that guide the proposed research. Finally, the combination of the assembled expertise, adequate facilities, strong collaborations and institutional support will allow us to conduct this study in a research environment that is conducive to its success.