Role of Reishi on cell surface proteins and signaling modulation in IBC

灵芝对 IBC 细胞表面蛋白和信号调节的作用

• 批准号: 8459318
• 负责人: Ivette Suarez
• 金额: $ 3.09万
• 依托单位: UNIVERSIDAD CENTRAL DEL CARIBE
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-09-26 至 2015-09-25
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8459318
• 关键词: Agaricales; Age of Onset; Amino Acids; Biomedical Research; Breast Cancer Cell; Bromodeoxyuridine; Cancer Patient; Cancer cell line; Cancerous; Cell Culture Techniques; Cell Membrane Proteins; Cell Proliferation; Cell Surface Proteins; Cell Survival; Cell membrane; Cell surface; Cell-Matrix Junction; Cells; Chinese Traditional Medicine; Data; Dermal; Development; Diagnosis; Disease; Dissection; Distant Metastasis; E-Cadherin; Early Diagnosis; Embolism; Epidermal Growth Factor; Epidermal Growth Factor Receptor; Epithelial Cells; Erlotinib; Event; Generations; Goals; Human; In Vitro; Inflammatory; Invaded; Knowledge; Label; Laboratories; Lymphatic System; MCF10A cells; Malignant Neoplasms; Mammary gland; Mass Spectrum Analysis; Medical; Membrane Proteins; Minority; Mission; Molecular; Molecular Biology Techniques; National Research Service Awards; Neoplasm Metastasis; Operative Surgical Procedures; Pathway interactions; Phenotype; Protein Inhibition; Proteins; Proteome; Proteomics; Proto-Oncogene Proteins c-akt; Publishing; Radiation; Radiation therapy; Reishi mushroom; Research; Resistance; Role; SCID Mice; Signal Pathway; Signal Transduction; Stable Isotope Labeling; Students; Survival Rate; Testing; Therapeutic; Therapeutic Agents; Training; Tumor Weights; United States National Institutes of Health; Vascular System; Work; anti-cancer therapeutic; base; career; combat; design; differentiation-stimulating factor; human tissue; in vivo; inflammatory breast cancer; inhibitor/antagonist; lapatinib; lymph nodes; malignant breast neoplasm; matrigel; molecular marker; mortality; neoplastic cell; overexpression; pre-doctoral; prevent; programs; protein expression; research study; stem; tumor; tumor growth; tumor progression; vasculogenesis

DESCRIPTION (provided by applicant): Inflammatory Breast Cancer (IBC) is one of the most lethal forms of breast cancer, where patients show less than a five percent survival rate beyond five years when treated with surgery or radiation therapy. IBC cells secrete differentiation factors, stimulate vasculogenesis, and invade as tumor emboli located within a de novo formed vessel. In IBC, the embolus maintains cell-cell attachments as it moves through the vessel and lodges within a dermal lymph node, causing the IBC inflammatory phenotype. Although the molecular changes that dictate IBC invasion remain to be fully elucidated, IBC cell lines and human tissues, overexpress E-cadherin, Epidermal Growth Factor Receptor (EGFR), and Human Epidermal Growth Factor 2 (HER2) which are cell surface proteins associated with maintaining tumor spheroid integrity, increase IBC tumor growth rate, and invasion and metastasis. Since IBC tumor cell emboli are more efficient at forming metastases and are more resistant to chemo and radiotherapy than single cells, it seems feasible to prevent IBC with a compound that has the ability to disintegrate the cell spheroids. We recently published that Ganoderma lucidum (Reishi), a mushroom used in Traditional Chinese Medicine, selectively inhibits IBC cell viability, but not viability of non- cancerous mammary epithelial cells. Also, we have shown that Reishi inhibits the expression of key proteins important in IBC progression, and tumor spheroid fomation. The main hypothesis of the present proposal is that Reishi inhibits cell surface protein expression, which contributes to disintegration of the tumor spheroids that are created during IBC progression, resulting in invasion inhibition combined with altered intracellular signaling. In support of this hypothesis, preliminary studies in our laboratory showe that Reishi 1) downregulates a number of cancer promoting molecules, 2) Reishi inhibits EGFR and E-cadherin protein abundance in vitro and in vivo. 3) Reishi inhibits IBC tumor growth and tumor weight in severe combined immunodeficient (SCID) mice by 58% and 45%, respectively. We propose that Reishi downregulates the expression of cell surface proteins involved in the progression of IBC and chemosensitize the IBC cells to EGFR/HER2 therapy treatment. Therefore, the main goal of this proposal is to validate the inhibitory role of Reishi on IBC progression via the inhibition of plasma membrane (PM) proteins and subsequent EGFR intracellular signaling events. To test our hypothesis, we propose 1) to isolate and identify PM proteins in non- cancerous mammary epithelial cells and in IBC cells treated with Reishi and 2) to investigate the therapeutic potential of Reishi in IBC cells, focusing on EGFR/HER2 signaling cascades. The proposed dissection of the molecular mechanisms of IBC is expected to significantly advance the current understanding and development of targeted therapeutic agents for IBC as well as contribute to a rapid and accurate diagnosis of this mortal disease. PUBLIC HEALTH RELEVANCE: Inflammatory Breast Cancer (IBC) is a rare and aggressive form of invasive breast cancer and it is characterized by rapid progression, local and distant metastases, younger age of onset, and lower overall survival compared with other breast cancers. The accurate and early diagnosis of IBC and the development of targeted therapy against this deadly disease remain a great medical challenge. The proposed dissection of the molecular mechanisms of IBC is expected to significantly advance the current understanding and development of targeted therapeutic agents for IBC as well as contribute to a rapid and accurate diagnosis of this mortal disease.

描述（由申请人提供）：炎症性乳腺癌（IBC）是最致命的乳腺癌形式之一，在接受手术或放射治疗治疗时，患者的存活率少于5％。 IBC细胞分泌分化因子，刺激血管生成，并作为位于从头形成的血管内的肿瘤栓子入侵。在IBC中，栓子通过血管移动并在真皮淋巴结中养殖时保持细胞 - 细胞附着，从而导致IBC炎症表型。尽管决定IBC侵袭的分子变化尚未充分阐明，但IBC细胞系和人体组织，过表达的E-钙粘着蛋白，表皮生长因子受体（EGFR）和人类表皮生长因子2（HER2）是与细胞表面蛋白相关的细胞表面蛋白，与维持肿瘤完整性，IBC肿瘤的生长速度和Invenasis保持相关。由于与单个细胞相比，IBC肿瘤细胞栓塞在形成转移方面更有效，对化学和放射疗法具有更大的耐药性，因此可以防止具有能够崩解细胞球体的化合物的IBC。我们最近发表了说，一种用于传统中药的蘑菇Ganoderma Lucidum（Reishi）有选择地抑制IBC细胞的生存力，但不是非癌性乳腺上皮细胞的生存能力。另外，我们 已经表明，灵芝抑制了在IBC进展中重要的关键蛋白的表达和肿瘤球体的效果。本提案的主要假设是雷希氏抑制细胞表面蛋白的表达，这有助于在IBC进展过程中产生的肿瘤球体分解，从而导致侵袭抑制与细胞内信号的改变。为了支持这一假设，我们的实验室中的初步研究表明了Reishi 1）下调了许多促进分子的癌症，2）Reishi抑制EGFR和E-Cadherin蛋白在体外和体内的丰度​​。 3）Reishi分别抑制严重的免疫缺陷（SCID）小鼠的IBC肿瘤生长和肿瘤的体重分别为58％和45％。我们提出，灵芝下调了参与IBC进展的细胞表面蛋白的表达，并使IBC细胞对EGFR/HER2治疗的化学敏感性。因此，该提案的主要目标是通过抑制质膜（PM）蛋白以及随后的EGFR细胞内信号传导事件来验证Reishi对IBC进展的抑制作用。为了检验我们的假设，我们建议1）分离和鉴定非癌性乳腺上皮细胞中的PM蛋白以及用Reishi处理的IBC细胞和2）研究Reishi在IBC细胞中的治疗潜力，重点是EGFR/HER2信号级联。提议的IBC分子机制的解剖有望显着提高IBC靶向治疗剂的当前理解和发展，并有助于快速准确诊断这种致命疾病。 公共卫生相关性：炎症性乳腺癌（IBC）是一种罕见且具有侵略性的侵入性乳腺癌形式，其特征是快速进展，局部和遥远的转移，年轻的发病年龄和与其他乳腺癌相比较低。 IBC的准确和早期诊断以及针对这种致命疾病的有针对性疗法的发展仍然是一个巨大的医学挑战。提议的IBC分子机制的解剖有望显着提高IBC靶向治疗剂的当前理解和发展，并有助于快速准确诊断这种致命疾病。