Molecular mechanism of bitter melon juice efficacy against pancreatic cancer.

苦瓜汁抗胰腺癌的分子机制。

• 批准号: 9326951
• 负责人: Rajesh Agarwal
• 金额: $ 38.28万
• 依托单位: UNIVERSITY OF COLORADO DENVER
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-09-01 至 2019-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9326951
• 关键词: Adenocarcinoma; Adverse effects; Affect; Anti-Inflammatory Agents; Anti-inflammatory; Apoptosis; Apoptotic; Asians; Automobile Driving; Biological Markers; CASP3 gene; Cancer Etiology; Cancer Model; Cancer Patient; Cancer Relapse; Cell Culture Techniques; Cell Line; Cellular Metabolic Process; Cessation of life; Clinical Trials; Complex; Country; Data; Diagnosis; Disease; Dose; Electron Transport; Enzymes; Erinaceidae; Event; FRAP1 gene; Food; Generations; Glycolysis; Growth; Health; Health Benefit; Human; Incidence; Inhibition of Apoptosis; Juice; Life; Malignant Neoplasms; Malignant neoplasm of pancreas; Mediating; Melons; Membrane Potentials; Metabolic; Metabolism; Mitochondria; Molecular; Mus; Mutation; NOD/SCID mouse; Neoplasm Metastasis; Nude Mice; Nutrient; Oral; Outcome; Oxidative Phosphorylation; Pancreas; Pathway interactions; Population; Process; Property; Publishing; Reactive Oxygen Species; Reporting; Resistance; Role; Safety; Signal Pathway; Signal Transduction; Signaling Molecule; Small Interfering RNA; Stress; Survival Rate; Symptoms; Testing; Time; Toxic effect; Transgenic Mice; Vegetables; Xenograft Model; Xenograft procedure; base; cancer cell; cancer stem cell; chemotherapy; diabetic; experimental study; feeding; gemcitabine; glucose metabolism; glucose uptake; in vivo; inhibitor/antagonist; mTOR Signaling Pathway; member; mitochondrial membrane; mortality; mouse model; notch protein; novel; pancreatic cancer cells; public health relevance; response; self-renewal; smoothened signaling pathway; translational impact; tumor; tumor progression; tumor xenograft

DESCRIPTION (provided by applicant): Pancreatic cancer (PanC) is an aggressive disease; median life of PanC patients post-diagnosis is <6 months and overall 5-year survival rate is 3-5%. Gemcitabine is the frontline chemotherapy for PanC that effectively eliminates bulk PanC cells; however spares cancer stem cell (CSC) population which causes cancer relapse as well as aggressiveness. Together, it is clear that additional strategies are urgently warranted to effectively lower PanC incidence, target CSC to control PanC relapse, and associated mortality. Noteworthy, PanC is a complex disease with multiple combinations of mutations, and therefore it is necessary to identify the agents with multiple targets to control both PanC growth and CSC-associated PanC relapse. Our published and preliminary studies show that bitter melon (Momordica charantia) juice (BMJ) significantly decreases the viability and induces strong apoptotic death of human PanC cell lines, which was associated with a robust AMPK activation, as both AMPK inhibitor and siRNA reversed BMJ-caused apoptosis. Furthermore, BMJ inhibited glycolysis and oxidative phosphorylation rate in PanC cells. Together these results suggested a 'metabolic shift' by BMJ in PanC cells. BMJ also strongly inhibited the sphere formation by PanC CSCs suggesting that it also targets CSC for its anti-PanC efficacy. Most notably, BMJ feeding by oral gavage at only 5mg dose/mouse/day resulted in 60% inhibition in MIA PaCa-2 xenograft growth in nude mice without any noticeable side effects or toxicity. Immunohistochemical analysis of xenografts showed that BMJ also inhibits proliferation and CSC biomarkers, induces apoptosis, and activates AMPK in vivo. Together, based on these and other findings, we hypothesize that BMJ causes metabolic shift in PanC cells through nutrient stress, AMPK activation and inhibiting signaling molecules related to metabolism and proliferation, which leads to strong growth inhibition and apoptosis specifically in PanC cells. Additionally, BMJ targets Notch/ Hedgehog signaling to effectively eliminate PanC CSC population; resulting in strong activity against PanC. The specific aims proposed are: I) To further define the mechanisms by which BMJ targets metabolism and affects AMPK-mediated growth inhibition and apoptosis in PanC cells; II) To further define the mechanisms by which BMJ targets Notch and Hedgehog pathways and effectively inhibits CSC population in PanC; and III) To further establish BMJ molecular mechanisms defined in specific aims I and II in PDX1-Cre; LSL-KRASG12D transgenic mouse model. It is important to highlight here that bitter melon is widely consumed as vegetable as well as juice especially in Asian countries; and has been attributed with multiple health beneficial properties such as anti-diabetic, anti-inflammatory, etc. Most notably, bitter melon has been tested in several clinical trials for its anti-diabetic effects and has plenty of human safety data. We, therefore, anticipate that the positive outcomes from the proposed studies will provide compelling rationale for initiating clinical trials to establish BMJ activity against human pancreatic cancer.

描述（由申请人提供）：胰腺癌（PanC）是一种侵袭性疾病； PanC 患者诊断后的中位寿命<6 个月，总体 5 年生存率为 3-5%。吉西他滨是 PanC 的一线化疗药物，可有效消除大量 PanC 细胞；然而，癌症干细胞（CSC）群体却不会受到影响，从而导致癌症复发和侵袭性。总之，很明显，迫切需要采取其他策略来有效降低 PanC 发病率，以 CSC 为目标来控制 PanC 复发和相关死亡率。值得注意的是，PanC 是一种具有多种突变组合的复杂疾病，因此有必要鉴定具有多个靶点的药物来控制 PanC 生长和 CSC 相关 PanC 复发。我们发表的初步研究表明，苦瓜（Momordica charantia）汁（BMJ）显着降低人类 PanC 细胞系的活力并诱导强烈的凋亡性死亡，这与 AMPK 的强烈激活有关，因为 AMPK 抑制剂和 siRNA 都逆转了 BMJ-引起细胞凋亡。此外，BMJ 抑制 PanC 细胞中的糖酵解和氧化磷酸化速率。这些结果共同表明 BMJ 在 PanC 细胞中发生了“代谢转变”。 BMJ 还强烈抑制 PanC CSC 的球体形成，表明它还针对 CSC 发挥抗 PanC 功效。最值得注意的是，仅以 5 毫克剂量/小鼠/天口服 BMJ 喂养，即可对裸鼠中的 MIA PaCa-2 异种移植物生长产生 60% 的抑制，且没有任何明显的副作用或毒性。异种移植物的免疫组织化学分析表明，BMJ 还在体内抑制增殖和 CSC 生物标志物、诱导细胞凋亡并激活 AMPK。总之，基于这些和其他发现，我们假设 BMJ 通过营养应激、AMPK 激活和抑制与代谢和增殖相关的信号分子引起 PanC 细胞的代谢变化，从而导致强烈的生长抑制和凋亡，特别是在 PanC 细胞中。此外，BMJ 以 Notch/ Hedgehog 信号为目标，有效消除 PanC CSC 群体；导致针对 PanC 的强烈活动。提出的具体目标是： I) 进一步明确 BMJ 靶向代谢并影响 PanC 细胞中 AMPK 介导的生长抑制和凋亡的机制； II) 进一步明确BMJ靶向Notch和Hedgehog通路并有效抑制PanC中CSC群体的机制； III) 进一步建立 PDX1-Cre 中特定目标 I 和 II 定义的 BMJ 分子机制； LSL-KRASG12D 转基因小鼠模型。这里需要强调的是，苦瓜作为蔬菜和果汁被广泛消费，尤其是在亚洲国家；并被认为具有多种有益健康的特性，如抗糖尿病、抗炎等。最值得注意的是，苦瓜具有 其抗糖尿病作用已通过多项临床试验测试，并且具有足够的人体安全性 数据。因此，我们预计拟议研究的积极结果将为启动临床试验以确定 BMJ 对抗人类胰腺癌的活性提供令人信服的理由。