Mechanistic Studies on Prostate Cancer Prevention by Gugulipid

古古脂预防前列腺癌的机制研究

• 批准号: 8542782
• 负责人: Dong Xiao
• 金额: $ 28.77万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 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/8542782
• 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

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，通常在生命的第六或第七个十年被诊断出来，这为干预提供了很大的机会窗口，以预防或减缓疾病的进展。因此，非常需要从这些天然产物中临床开发相对安全但可延迟人前列腺癌发作和/或进展的药剂。Gugulipid（GL）是Commiphora mukul的提取物，已在印度阿育吠陀医学中安全使用数千年，用于治疗不同的疾病。GL已被用于许多临床试验，这些临床试验侧重于其降胆固醇作用，并且没药标准化制剂的产品已经在美国作为降胆固醇剂用于人类。在R21 CA 143104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。GL处理人前列腺癌LNCaP（雄激素依赖型）及其雄激素非依赖型C81细胞，在可达浓度（~ 1.5mol/L）时可诱导细胞凋亡，从而降低细胞活力。有趣的是，正常人前列腺上皮细胞系（PrEC）对GL的生长抑制和凋亡诱导具有显著更强的抗性，这是潜在癌症预防剂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周龄开始每周三次经口灌胃GL，持续20周，可显著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，而不会导致体重减轻或任何其他副作用。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氨基末端激酶（JNK）的激活有关。目的/假设：我们假设GL选择性杀死前列腺癌细胞可能靶向ROS介导的机制。本R 01申请建议进行临床前研究，以确定GL（一种补充和替代药物）在体内预防前列腺癌的疗效，并阐明其抗癌作用的机制。具体目标1-3中的拟定研究将确定GL引起人前列腺癌细胞凋亡和自噬的机制。此外，具体目标4的研究将评价GL预防前列腺癌的体内疗效和机制。具体目标：为了验证我们的假设，我们提出了以下具体目标：1。使用LNCaP、C81和PC-3人前列腺癌细胞以及人正常前列腺上皮细胞PrEC作为模型，确定GL对癌细胞的选择性杀伤是否通过ROS介导的机制。所提出的实验将测试人前列腺癌细胞中GL介导的细胞凋亡是否依赖于ROS;此外，是否来自ESTA的ROS用于启动这种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诱导的自噬和凋亡之间的串扰及其机制。这些研究将确定GL诱导自噬的效果，以及GL介导的人前列腺癌细胞自噬是否涉及线粒体ROS的调节。此外，GL引起的自噬在GL诱导的细胞凋亡中的真实的作用将被研究。 3.使用LNCaP、PC-3和PrEC作为模型，确定GL对细胞凋亡和自噬的同时诱导是否受到JNK信号传导轴的ROS依赖性调节的调节。本研究旨在阐明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在TRAMP小鼠中预防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和前列腺癌的发生率、转移和负荷的疗效。此外，我们将评价GL诱导的细胞凋亡和自噬的相关生物标志物（具体目的1-3），以使用溶媒（对照）和GL处理小鼠的肿瘤组织测定细胞凋亡以及细胞凋亡和自噬调节蛋白的水平。创新与意义：我们相信，我们的建议福尔斯属于创新研究的标准，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，并使用TRAMP转基因小鼠确定GL预防前列腺癌发生的有效性。GL是一种有前途的化合物，由于其对前列腺癌具有选择性抗癌活性，因此在预防应用中具有显著的潜力。GL具有立即临床转化的潜力，因为膳食补充剂已经在美国用于人类健康益处，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以及优化基于GL的前列腺癌预防和治疗方案中有用。