Anti-tumor properties of curcumin in oral cancers: role of thioredoxin reductase

姜黄素在口腔癌中的抗肿瘤特性：硫氧还蛋白还原酶的作用

• 批准号: 7738540
• 负责人: Stephen William Tuttle
• 金额: $ 17.32万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-05-21 至 2011-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7738540
• 关键词: 3' Untranslated Regions; Adenovirus Vector; Affect; Alcohol consumption; Animal Model; Antioxidants; Apoptosis; Apoptotic; Biological Assay; Biological Availability; Biological Factors; Blood Circulation; C-terminal; Cancer cell line; Cell Death; Cell Line; Cells; Cervical; Cessation of life; Chemicals; Chemoprevention; Chemopreventive Agent; Clinical Trials; Covalent Interaction; Curcuma longa; Curcumin; Cysteine; DNA Insertion Elements; DNA Repair; Data; Diagnosis; Diet; Dominant-Negative Mutation; Dose; Electrons; Enzymes; Genes; Genetic Transcription; Genus Cola; Growth; Head and Neck Cancer; Head and Neck Squamous Cell Carcinoma; Human; Hydrogen Peroxide; Implant; In Vitro; Ionizing radiation; Knock-in Mouse; Length; Lesion; Luciferases; Malignant Epithelial Cell; Malignant Neoplasms; Measures; Mediating; Mediator of activation protein; Modification; Morbidity - disease rate; NADP; NADPH Oxidase; Nude Mice; Operative Surgical Procedures; Oral; Oral Leukoplakia; Oxidation-Reduction; Pharmaceutical Preparations; Phase I Clinical Trials; Plant Roots; Plants; Plasmid Cloning Vector; Plasmids; Premalignant; Property; Proteins; Protocols documentation; Radiation; Radiation Tolerance; Radiation therapy; Radiation-Sensitizing Agents; Reaction; Reporter; Resistance; Rhizome; Risk Factors; Role; Scheme; Second Primary Cancers; Selenocysteine; Signaling Protein; Smoke; Smoking; Subcutaneous Tissue; Superoxides; Survival Rate; TP53 gene; Testing; Therapeutic; Thioredoxin; Toxic effect; Transcription Factor AP-1; Transfection; Translating; Work; adduct; angiogenesis; bioactive food component; cancer cell; cell growth regulation; chemotherapy; cytotoxic; expression vector; gene cloning; head and neck cancer patient; in vivo; inhibitor/antagonist; keratinocyte; knock-down; malignant mouth neoplasm; mouse model; neoplastic cell; outcome forecast; overexpression; pre-clinical; public health relevance; radiation resistance; research study; response; selenoprotein; small hairpin RNA; thioredoxin reductase; thioredoxin reductase 1; tumor; tumor growth

DESCRIPTION (provided by applicant): Thioredoxin reductase 1 (TxRd1), is an essential selenoprotein with defined roles in redox regulation of cell growth, death (apoptosis), transcription, DNA repair, and angiogenesis. TxRd1 is overexpressed in many human cancers and these elevated levels of TxRd1 have been associated with poor prognosis due to enhanced cellular resistance to chemo- and radiation therapy. Chemical inhibitors of TxRd have yielded promising results in preclinical and clinical trials as cancer therapeutics and radiation sensitizers. Curcumin is a naturally occurring plant polyphenolic antioxidant with potent chemopreventive and antintumorigenic properties in preclinical animal models and a demonstrated low toxicity profile in Phase I clinical trials Recent data demonstrate a covalent interaction between curcumin and the conserved c-terminal cys-x-sec-cys motif in TxRd1. resulting in inhibition ofthe catalytic interaction between TxRd and thioredoxin. The electrons donated by NADPH are instead diverted to O2, to form superoxide and ultimately hydrogen peroxide. In this proposal we will investigate the role of TxRd1 as a critical mediator of the antitumorigeniceffects of curcumin alone and in combination with ionizing radiation in oraopharyngeal cancer cell lines, in vitro and in vivo. . Our preliminary data demonstrated > 50 fold higher level of TxRd1 in the human squamous carcinoma cell line FaDu compared to either an oral leukoplakia cell line MSK-Leuk1 or nontransformed keratinocytes. In specific aim 1, we plan to knockdown TxRd1 levels in FaDu cells using shRNA to the 3'UTR of the human TxRd1 gene cloned into the pSilencer expression vector. Conversely, TxRd1 will be overexpressed in MSK-leuk1 cells by transfection with the full length wild type TxRd1 gene, including the selenocysteine insertion element, from the 3'UTR. The stable clones selected above will be used to determine whether TxRd1 mediates the anti-proliferative and pro-apoptotic actions of curcumin, in vitro. In specific aim 2, we will use the stable clones selected above, to determine whether curcumin's efficacy as a radiation sensitizer also depends on inhibition of TxRd1. In specific aim 3, the TxRd1 knockdown and knock in cell lines, and the parental cell lines, will be grown as tumors in the cervical subcutaneous tissue of athymic nude mice. The efficacy of curcumin administered in the diet alone or in combination with ionizing radiation on orthotopic tumor growth and tumor cell death will be measured. Since curcumin is a natural bioactive food component with no overt systemic toxicities, promising results from these preclinical experiments can be readily translated into a chemoprevention schemes or combined chemo-radiation therapy protocol for treating head and neck cancers. PUBLIC HEALTH RELEVANCE: Head and neck cancers (HNC) are the sixth most common form of cancer worldwide, with over 500,000 new cases diagnosed annually. Although advances in surgery, and combined of radio and chemotherapies, have decreased morbidity, the survival rate at 5 years after initial prognosis remains at < 50%. Moreover, since smoking and alcohol use are the two most prevalent risk factors associated with HNSCC, field cancerization leads to second primary cancers in 20% of HNC patients. Cucumin, a natural product derived from the root of the plant Curcuma longa, has shown promise as a cancer chemopreventive, chemotherapeutic and radiation sensitizer, in preclinical and phase 1 clinical trials. The primary drawback to the use of curcumin is limited bioavailability due to low concentrations of the drug reaching the circulation. However, curcumin has been effective in tumors, such as colon and HNC, where direct access to the drug is possible. The primary aim of this study is to develop curcumin as a chemotherapeutic and radiation sensitizer for treating HNC..

描述（由申请人提供）：硫氧还蛋白还原酶1（TXRD1）是一种必不可少的硒蛋白，在细胞生长，死亡（凋亡），转录，DNA修复和血管生成的氧化还原调节中具有明确的作用。 TXRD1在许多人类癌症中过表达，由于细胞对化学和放射疗法的耐药性增强，TXRD1水平升高与预后不良有关。 TXRD的化学抑制剂在临床前试验和临床试验中作为癌症治疗和辐射敏化剂产生了有希望的结果。姜黄素是一种天然存在的植物多酚抗氧化剂，在临床前动物模型中具有有效的化学预防和抗肿瘤性特性，在I期临床试验中显示出低毒性的毒性较低，显示了姜黄素与保守的C-terminal Cys-terminal Cys-x-x-cys-x-cys-cys-cys-cys-cys-cys-cys-cys in txrd1。导致TXRD和硫氧还蛋白之间的催化相互作用抑制。相反，由NADPH捐赠的电子被转移到O2，形成超氧化物并最终形成过氧化氢。在该提案中，我们将研究TXRD1作为姜黄素抗肿瘤胶质素的关键介体的作用，并在体外和体内与咽咽癌细胞系中的电离辐射结合使用。 。我们的初步数据证明，与口服白细胞细胞系MSK-Leuk1或未转化的角质形成细胞相比，人类鳞状癌细胞系Fadu中TXRD1的水平> 50倍。在特定的目标1中，我们计划使用shRNA在FADU细胞中敲低TXRD1水平，以克隆到Psilencer表达载体中的人类TXRD1基因的3'UTR。相反，TXRD1通过从3'UTR转染，包括全长野生型TXRD1基因（包括硒代半胱氨酸插入元件），在MSK-Leuk1细胞中过表达TXRD1。上面选择的稳定克隆将用于确定TXRD1是否介导了姜黄素的抗增殖和促凋亡作用。在特定目标2中，我们将使用上面选择的稳定克隆来确定姜黄素作为辐射敏化剂的功效是否还取决于抑制TXRD1。在特定的目标3中，TXRD1敲低和细胞系中的敲击和亲本细胞系将作为腹膜裸鼠的宫颈皮下组织中的肿瘤生长。仅在饮食中施用姜黄素的功效，或与原位肿瘤生长和肿瘤细胞死亡的电离辐射结合使用。由于姜黄素是一种天然的生物活性食品成分，没有明显的全身毒性，因此这些临床前实验的有希望的结果很容易被转化为化学预防方案或用于治疗头颈部癌症的化学预防疗法。公共卫生相关性：头颈癌（HNC）是全球第六种最常见的癌症形式，每年诊断出500,000多个新病例。尽管手术的进展以及无线电和化学疗法的结合降低了发病率，但初始预后后5年的存活率仍<50％。此外，由于吸烟和饮酒是与HNSCC相关的两个最普遍的危险因素，因此在20％的HNC患者中，现场癌化会导致第二个原发性癌症。在临床前和1期临床试验中，cucumin是一种源自植物姜黄的根源的天然产物，它作为一种癌症化学预防，化学治疗和辐射敏化剂表现出了希望。使用姜黄素的主要缺点是由于低浓度的药物到达循环的浓度而有限的生物利用度。但是，姜黄素在肿瘤（例如结肠和HNC）中有效，在这些肿瘤中可以直接进入该药物。这项研究的主要目的是开发姜黄素作为治疗HNC的化学治疗和辐射敏化剂。