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: 9134970
• 负责人: Ivette Suarez
• 金额: $ 1.8万
• 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 2016-04-25
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9134970
• 关键词: Age of Onset; Amino Acids; Biomedical Research; Breast Cancer Cell; Breast Cancer Patient; Breast Cancer cell line; Breast Epithelial Cells; Bromodeoxyuridine; 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; Disease; Dissection; Distant Metastasis; E-Cadherin; Early Diagnosis; Embolism; Epidermal Growth Factor; Epidermal Growth Factor Receptor; Erlotinib; Event; Generations; Goals; Human; In Vitro; Inflammatory; Invaded; Knowledge; Label; Laboratories; Lymphatic System; MCF10A cells; Malignant Neoplasms; 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; accurate diagnosis; 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; rapid diagnosis; research study; stem; targeted treatment; 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.

描述（由申请人提供）：炎性乳腺癌 (IBC) 是最致命的乳腺癌形式之一，患者在接受手术或放射治疗后五年后的存活率不到 5%。 IBC 细胞分泌分化因子，刺激血管发生，并作为肿瘤栓子侵入新形成的血管内。在 IBC 中，栓子在穿过血管并停留在真皮淋巴结内时保持细胞与细胞的附着，从而导致 IBC 炎症表型。尽管决定 IBC 侵袭的分子变化仍有待完全阐明，但 IBC 细胞系和人体组织过度表达 E-钙粘蛋白、表皮生长因子受体 (EGFR) 和人表皮生长因子 2 (HER2)，这些细胞表面蛋白与维持肿瘤球体完整性相关，增加 IBC 肿瘤生长速率以及侵袭和转移。由于 IBC 肿瘤细胞栓子比单个细胞更能有效地形成转移，并且对化疗和放疗具有更强的抵抗力，因此使用能够分解细胞球体的化合物来预防 IBC 似乎是可行的。我们最近发表了灵芝（灵芝），一种用于中药的蘑菇，选择性抑制 IBC 细胞活力，但不抑制非癌性乳腺上皮细胞的活力。另外，我们 研究表明，灵芝可抑制 IBC 进展和肿瘤球体形成中重要的关键蛋白的表达。本提案的主要假设是灵芝抑制细胞表面蛋白表达，这有助于 IBC 进展过程中产生的肿瘤球体的崩解，从而导致侵袭抑制和细胞内信号传导改变。为了支持这一假设，我们实验室的初步研究表明，灵芝 1) 下调许多促癌分子，2) 灵芝在体外和体内抑制 EGFR 和 E-钙粘蛋白丰度。 3) 灵芝可抑制严重联合免疫缺陷 (SCID) 小鼠 IBC 肿瘤生长和肿瘤重量，分别达 58% 和 45%。我们建议灵芝下调参与 IBC 进展的细胞表面蛋白的表达，并使 IBC 细胞对 EGFR/HER2 疗法化疗敏感。因此，该提案的主要目标是通过抑制质膜（PM）蛋白和随后的 EGFR 细胞内信号事件来验证灵芝对 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和用灵芝处理的 IBC 细胞中的 PM 蛋白，2) 研究灵芝在 IBC 细胞中的治疗潜力，重点关注 EGFR/HER2 信号级联。所提出的对 IBC 分子机制的剖析有望显着推进目前对 IBC 靶向治疗药物的理解和开发，并有助于快速、准确地诊断这种致命疾病。