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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60或70岁时被诊断出来，这为预防或减缓疾病进展的干预提供了很大的机会窗口。因此，从这些天然产物中开发相对安全但能延缓人类前列腺癌发病和/或进展的药物是非常可取的。古古脂（GL）是印度阿育吠陀医学中安全使用了数千年的草药，用于治疗不同的疾病。GL已被用于许多临床试验，这些试验的重点是它的降胆固醇效果，而Commiphora mukul的标准化配方产品已经在美国作为降胆固醇剂用于人体。在R21 CA143104的支持下，我们首次在体外和体内研究了GL（美国专利产品）在前列腺癌中的抗肿瘤活性。研究发现，在药理学上可达到的浓度（~ 15mol /L）下，GL可诱导人前列腺癌LNCaP（雄激素依赖性）及其雄激素非依赖性变体C81细胞凋亡，从而降低细胞活力。有趣的是，正常的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更强的抵抗力，这是潜在癌症预防药物的一个非常理想的特征。最重要的是，我们的初步研究表明，从5周龄开始，每周灌胃3次GL，持续20周，可以显著抑制小鼠前列腺（TRAMP）转基因腺癌的发生和进展，而不会造成体重减轻或任何其他副作用。我们的初步研究也表明，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）的激活有关。目的/假设：我们假设gl选择性杀伤前列腺癌细胞可能是针对ros介导的机制。本r01申请拟开展临床前研究，确定补充替代药物GL在体内预防前列腺癌的疗效，阐明其抗癌作用机制。在特定目标1-3中提出的研究将确定GL导致人类前列腺癌细胞凋亡和自噬的机制。此外，特异性靶4的研究将评估GL预防前列腺癌的体内疗效和机制。具体目的：为了检验我们的假设，我们提出以下具体目的：1。以LNCaP、C81、PC-3人前列腺癌细胞以及人正常前列腺上皮细胞PrEC为模型，确定GL是否通过ros介导的机制选择性杀伤癌细胞。本实验拟检测gl介导的人前列腺癌细胞凋亡是否依赖于ROS；此外，线粒体来源的ROS是否有助于启动这种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诱导的自噬与凋亡之间的串扰及其机制。拟开展的研究将确定GL诱导自噬的作用，以及GL介导的人前列腺癌细胞自噬是否参与线粒体ROS的调节。此外，还将探讨GL诱导的自噬在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ros依赖性JNK信号轴调控。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是如何引发癌细胞死亡的？哪个在ROS的下游？4. 确定GL在小鼠体内预防前列腺癌发生转移的作用及机制。我们将采用TRAMP小鼠模型，确定口服GL对PIN和前列腺癌的发生率、转移和负担的影响。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（特异性目的1-3），使用小鼠肿瘤组织（对照）和GL检测细胞凋亡和细胞凋亡和自噬调节蛋白的水平。创新与意义：我们首次提出探讨GL预防前列腺癌的作用和分子机制，并利用TRAMP转基因小鼠确定GL预防前列腺癌的功效，相信我们的提案属于创新研究的标准。GL对前列腺癌具有选择性抗癌活性，是一种很有前景的化合物，在预防前列腺癌方面具有重要的应用潜力。GL具有立即临床转化的潜力，因为一种膳食补充剂已经在美国用于人体健康，对人体没有任何毒性。确定GL诱导细胞凋亡机制的长期内在价值在于鉴定GL反应的生物标志物，这些标志物可能在未来的临床试验中有用，并有助于优化基于GL的前列腺癌预防和治疗方案。