Mechanistic Studies on Prostate Cancer Prevention by Gugulipid

古古脂预防前列腺癌的机制研究

基本信息

批准号：
8712195
负责人：
Shivendra Singh
金额：
$ 31万
依托单位：
UNIVERSITY OF PITTSBURGH AT PITTSBURGH
依托单位国家：
美国
项目类别：
财政年份：
2012
资助国家：
美国
起止时间：
2012-09-10 至 2015-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8712195
关键词：
Adenocarcinoma Adverse effects Age American Americas Androgens Apoptosis Autophagocytosis Ayurvedic Medicine Biological Assay Biological Factors Biological Markers Body Weight Body Weight decreased Cancer Cell Growth Cancer Etiology Cell Death Cell Line Cells Cessation of life Chemopreventive Agent Cholesterol Clinical Clinical Trials Commiphora mukul Complementary and alternative medicine DU145 Data Development Diagnosis Disease Progression Drug Formulations Epithelial Cells Future Generations Growth Health Benefit Human In Vitro Incidence Induction of Apoptosis Inhibition of Apoptosis Intervention Investigation JUN gene LNCaP Legal patent Life Malignant Neoplasms Malignant neoplasm of prostate Mediating Mitochondria Modeling Molecular Mus Neoplasm Metastasis Nude Mice Oral Population Prevention Prevention therapy Preventive Prostate Prostate carcinoma Proteins Reactive Oxygen Species Regimen Regulation Reporting Research Research Design Resistance Role Signal Transduction Testing Time Toxic effect Transgenic Mice Transgenic Organisms Translations Trust Tumor Tissue United States Variant Xenograft Model angiogenesis anticancer activity base cancer cell dietary supplements falls gugulipid in vivo innovation killings male men mortality mouse model novel novel strategies preclinical study prevent prostate cancer cell prostate cancer prevention prostate carcinogenesis research study response stress-activated protein kinase 1

项目摘要

DESCRIPTION (provided by applicant): Prostate cancer, a leading cause of cancer related deaths among men in the United States, is usually diagnosed in the sixth or seventh decades of life, which allows a large window of opportunity for intervention to prevent or slow progression of the disease. Thus, clinical development of agents from these natural products that are relatively safe but can delay onset and/or progression of human prostate cancer is highly desirable. Gugulipid (GL), extract of Commiphora mukul, has been safely used for thousands of years in the Indian Ayurvedic medicine to treat different ailments. GL has been used in many clinical trials that have focused on its cholesterol-lowering effect and the products of standardized formulations of Commiphora mukul are already in human use in U.S. as cholesterol-lowering agents. Supported by R21 CA143104, we, for the first time, have investigated the antitumor activity of GL (a US patent product) in prostate cancer in vitro and in vivo. GL treatment was found to decrease viability of human prostate cancer LNCaP (androgen-dependent) and its androgen-independent variant C81 cells by causing apoptosis induction at pharmacologically achievable concentrations (~1 5mol/L). Interestingly, a normal human prostate epithelial cell line (PrEC) is significantly more resistant to growth inhibition and apoptosis induction by GL, which is a highly desirable feature of potential cancer preventive agents. Most importantly, our preliminary studies showed that oral gavage of GL, thrice per week beginning at five weeks of age for 20 weeks, significantly inhibits prostate cancer incidence and progression in transgenic adenocarcinoma of mouse prostate (TRAMP) mice without causing weight loss or any other side effects. Our preliminary studies have also shown that the GL-induced cell death in prostate cancer cells was correlated with generation of reactive oxygen species (ROS) and activation of c-Jun NH2-terminal kinase (JNK). Objective/Hypothesis: we hypothesize that GL-selective killing of prostate cancer cells might target the ROS-mediated mechanism. This R01application proposes preclinical studies to determine the efficacy of GL, a Complementary and Alternative Medicine, for prevention of prostate cancer in vivo and to elucidate the mechanism of its anti-carcinogenic effect. The proposed studies in specific aims 1-3 will determine the mechanism(s) by which GL causes apoptosis and autophagy in human prostate cancer cells. Moreover, investigations of specific aim 4 will evaluate the in vivo efficacy and mechanisms of GL in prevention of prostate cancer. Specific Aims: To test our hypothesis, we propose the following specific aims: 1. Determine whether the selective killing of cancer cells by GL is through a ROS-mediated mechanism using LNCaP, C81 and PC-3 human prostate cancer cells as well as human normal prostate epithelial cell PrEC as a model. The proposed experiments will test whether the GL-mediated apoptosis in human prostate cancer cells is dependent on ROS; and furthermore, whether the mitochondria-derived ROS serve to initiate such GL-induced cell death. 2. Determine the crosstalk between GL-induced autophagy and apoptosis and its mechanisms using LNCaP, PC-3 and PrEC cells as well as PC-3 xenograft model. The propose studies will determine the effect of autophagy induction by GL, and whether GL-mediated autophagy in human prostate cancer cells involves regulation of mitochondrial ROS. Moreover, the real role of GL-caused autophagy in apoptosis induced by GL will be investigated. 3. Determine whether the simultaneous induction of apoptosis and autophagy by GL is regulated by the ROS-dependent regulation of JNK signaling axis using LNCaP, PC-3 and PrEC as a model. Studies are designed to elucidate the mechanism of simultaneous induction of autophagy and apoptosis by GL. Likewise, how does the GL-induced ROS trigger the cancer cell death? Which is downstream of ROS? 4. Determine in vivo efficacy and mechanism of GL for prevention of prostate carcinogenesis and metastasis in TRAMP mice. We will determine the efficacy of orally administered GL against the incidence, metastasis and burden of PIN and prostate carcinoma using the TRAMP mice model. Furthermore, we will evaluate the associated biomarkers of apoptosis and autophagy induced by GL (specific aims 1-3) to use the tumor tissues from the mice treated with vehicle (control) and GL for assay of apoptosis and levels of apoptosis and autophagy regulating proteins. Innovation and Significance: We trust that our proposal falls under the criteria of innovative research because we, for the first time, propose to probe into the effects and molecular mechanisms for prevention of prostate cancer by GL and to determine efficacy of GL for prevention of prostate carcinogenesis using TRAMP transgenic mice. GL is a promising compound and has significant potential in preventive applications since it has selectively anticancer activity for prostate cancer. GL has the potential for immediate clinical translation because a dietary supplement is already in human use for health benefits in America without any toxicity in humans. The long-term intrinsic value of defining the mechanism by which GL causes apoptosis induction resides in the identification of biomarkers of GL response potentially useful in future clinical trials and in the optimization of GL-based regimens for prevention and therapy of prostate cancer.

描述（由申请人提供）：前列腺癌是美国男性癌症相关死亡的主要原因，通常在六岁或七岁的时候被诊断出来，这为预防或减缓进展的干预提供了很大的机会的疾病。因此，非常需要临床开发来自这些天然产物的相对安全但可以延迟人类前列腺癌的发作和/或进展的药剂。 Gugulipid (GL) 是没药树 (Commiphora mukul) 的提取物，在印度阿育吠陀医学中已安全使用数千年，用于治疗不同的疾病。 GL 已用于许多临床试验，重点关注其降低胆固醇的作用，而没药标准化配方的产品已在美国作为降胆固醇剂用于人类。在R21 CA143104的支持下，我们首次研究了GL（美国专利产品）在体外和体内对前列腺癌的抗肿瘤活性。研究发现，GL 治疗可在药理学可达到的浓度 (~1·5mol/L) 下诱导细胞凋亡，从而降低人前列腺癌 LNCaP（雄激素依赖性）及其雄激素非依赖性变体 C81 细胞的活力。有趣的是，正常人前列腺上皮细胞系 (PrEC) 对 GL 的生长抑制和细胞凋亡诱导具有明显更强的抵抗力，这是潜在癌症预防剂非常理想的特征。最重要的是，我们的初步研究表明，从 5 周龄开始每周 3 次口服 GL，持续 20 周，可显着抑制转基因小鼠前列腺腺癌 (TRAMP) 小鼠的前列腺癌发病率和进展，且不会导致体重减轻或任何其他不良反应。副作用。我们的初步研究还表明，GL 诱导的前列腺癌细胞死亡与活性氧 (ROS) 的产生和 c-Jun NH2 末端激酶 (JNK) 的激活相关。目的/假设：我们假设 GL 选择性杀伤前列腺癌细胞可能针对 ROS 介导的机制。该 R01 申请提出了临床前研究，以确定 GL（一种补充和替代医学）在体内预防前列腺癌的功效，并阐明其抗癌作用的机制。具体目标 1-3 中拟议的研究将确定 GL 引起人类前列腺癌细胞凋亡和自噬的机制。此外，具体目标4的研究将评估GL预防前列腺癌的体内功效和机制。具体目标：为了检验我们的假设，我们提出以下具体目标： 1. 使用 LNCaP、C81 和 PC-3 人前列腺癌细胞以及人前列腺癌细胞，确定 GL 对癌细胞的选择性杀伤是否是通过 ROS 介导的机制。以正常前列腺上皮细胞PrEC为模型。拟议的实验将测试 GL 介导的人前列腺癌细胞凋亡是否依赖于 ROS；此外，线粒体衍生的 ROS 是否会引发 GL 诱导的细胞死亡。 2. 利用LNCaP、PC-3和PrEC细胞以及PC-3异种移植模型确定GL诱导的自噬和凋亡之间的串扰及其机制。拟议的研究将确定 GL 诱导自噬的效果，以及人类前列腺癌细胞中 GL 介导的自噬是否涉及线粒体 ROS 的调节。此外，还将研究 GL 引起的自噬在 GL 诱导的细胞凋亡中的真正作用。 3. 以LNCaP、PC-3和PrEC为模型，确定GL同时诱导细胞凋亡和自噬是否受到JNK信号轴ROS依赖性调节的调节。研究旨在阐明 GL 同时诱导自噬和凋亡的机制。同样，GL 诱导的 ROS 如何触发癌细胞死亡？ ROS 的下游是什么？ 4.确定GL预防TRAMP小鼠前列腺癌发生和转移的体内功效和机制。我们将使用 TRAMP 小鼠模型确定口服 GL 对 PIN 和前列腺癌的发病率、转移和负担的功效。此外，我们将评估GL诱导的细胞凋亡和自噬的相关生物标志物（具体目标1-3），以使用来自用载体（对照）和GL处理的小鼠的肿瘤组织来测定细胞凋亡以及细胞凋亡和自噬调节蛋白的水平。创新性和意义：我们相信我们的提案符合创新研究的标准，因为我们首次提出探讨GL预防前列腺癌的作用和分子机制，并确定GL预防前列腺癌的功效使用 TRAMP 转基因小鼠进行致癌作用。 GL 是一种很有前途的化合物，由于它对前列腺癌具有选择性抗癌活性，因此在预防应用中具有巨大潜力。 GL 具有立即临床转化的潜力，因为膳食补充剂已在美国用于人类健康益处，且对人类没有任何毒性。定义 GL 引起细胞凋亡诱导的机制的长期内在价值在于鉴定 GL 反应的生物标志物，这些生物标志物可能在未来的临床试验中有用，并优化基于 GL 的前列腺癌预防和治疗方案。