ONCOSTATIN M INDUCES VEGF IN HUMAN BREAST CARCINOMA CELLS

制瘤素 M 在人乳腺癌细胞中诱导 VEGF

• 批准号: 7959937
• 负责人: CHERYL LYNN JORCYK
• 金额: $ 9.48万
• 依托单位: UNIVERSITY OF IDAHO
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-04-01 至 2010-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7959937
• 关键词: Angiogenic Factor; Biological Assay; Breast Cancer Cell; Breast Carcinoma; Cells; Clinical; Coculture Techniques; Computer Retrieval of Information on Scientific Projects Database; Conditioned Culture Media; Cultured Cells; Data; Endothelial Cells; Enzyme-Linked Immunosorbent Assay; Epithelial Cells; Fatty acid glycerol esters; Funding; Grant; Granulocyte-Macrophage Colony-Stimulating Factor; Growth; Idaho; In Vitro; Institution; Laboratories; Lead; Light; Mammary Gland Parenchyma; Mammary gland; Matrix Metalloproteinases; Nude Mice; Production; Property; Research; Research Personnel; Resources; Signal Transduction; Source; System; Testing; United States National Institutes of Health; Vascular Endothelial Growth Factor A; Western Blotting; Whole Blood; angiogenesis; autocrine; cancer therapy; cell type; cytokine; in vivo; macrophage; malignant breast neoplasm; matrigel; neoplastic cell; neutralizing antibody; neutrophil; oncostatin M; paracrine; programs; tumor; tumor progression

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Oncostatin M (OSM) is a pleiotropic cytokine produced by many cell types, including neutrophils and tumor-associated macrophages. OSM inhibits the proliferation of breast cancer cells in vitro, and for this reason is being examined for its potential use in cancer treatment. Important to this endeavor are the results of preliminary studies done in our laboratory that show that OSM may promote pro-angiogenic factors in tumor cells, as is observed for endothelial cells. Additional studies show that OSM is expressed by tumor-associated neutrophils and breast cancer epithelial cells, but not by normal breast tissue. Furthermore, neutrophils isolated from whole blood do not express OSM until they are co-cultured with breast cancer cells. Our data demonstrates that OSM will stimulate breast cancer cells to produce angiogenesis-related matrix metalloproteinases (MMPs) and vascular endothelial growth factor-A (VEGF), which is an extremely potent pro-angiogenic factor. The implication of the latter finding is that while OSM may cause growth-arrest in breast cancer cells in vitro, it may also lead to the induction of angiogenesis in the tumor through production of VEGF. Such an induction of angiogenesis would counter the growth arrest properties of OSM by promoting angiogenesis-dependent breast cancer progression. In light of our findings, it is important to better understand the implications of VEGF induction to correctly evaluate the potential of OSM as a clinical cancer treatment. The Specific Aims of the proposal are: SPECIFIC AIM 1. DEMONSTRATE THAT NEUTROPHIL-DERIVED OSM WILL INDUCE VEGF FROM BREAST CANCER CELLS IN A PARACRINE FASHION. Neutrophils will secret OSM when they are co-cultured with breast cancer cells, but the pathophysiologic significance of this is unknown. Neutrophil-breast cancer cell co-cultures will be tested to determine whether neutrophil-secreted OSM will induce VEGF from breast cancer cells in a paracrine fashion. OSM and VEGF levels will be analyzed by ELISA. SPECIFIC AIM 2. IDENTIFY AND CHARACTERIZE THE SIGNAL THAT STIMULATES NEUTROPHILS TO RELEASE OSM. We will identify the signal generated by breast cancer cells that stimulates neutrophils to release OSM by performing Western blots/ELISAs on cell lysates/conditioned media of co-cultured cells treated with neutralizing antibodies to specific cytokines such as GM-CSF. In addition, we will determine whether neutrophil-breast cancer cell contact is needed for release of OSM by neutrophils in both a co-culture system (cell-contact) and a Transwell-culture (no cell contact). SPECIFIC AIM 3. INVESTIGATE THE ABILITY OF OSM-INDUCED VEGF TO STIMULATE ANGIOGENESIS AND PROMOTE BREAST CARCINOMA PROGRESSION IN VIVO. We will investigate whether OSM-induced VEGF will stimulate angiogenesis in vivo using the Matrigel plug assay, and assess whether OSM-transfected breast cancer cells injected into the cleared mammary fat pads of nude mice will promote tumor progression by stimulating VEGF-dependent angiogenesis in an autocrine fashion.

这个子项目是许多研究子项目中的一个 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH来源获得了主要资金， 因此可以在其他CRISP条目中表示。所列机构为 研究中心，而研究中心不一定是研究者所在的机构。 抑瘤素M（Oncostatin M，OSM）是由多种细胞类型产生的多效性细胞因子，包括中性粒细胞和肿瘤相关巨噬细胞。 OSM在体外抑制乳腺癌细胞的增殖，因此正在研究其在癌症治疗中的潜在用途。 我们实验室的初步研究结果表明，OSM可以促进肿瘤细胞中的促血管生成因子，正如在内皮细胞中观察到的那样。 其他研究表明，OSM由肿瘤相关中性粒细胞和乳腺癌上皮细胞表达，但不被正常乳腺组织表达。 此外，从全血中分离的中性粒细胞不表达OSM，直到它们与乳腺癌细胞共培养。 我们的数据表明，OSM将刺激乳腺癌细胞产生血管生成相关的基质金属蛋白酶（MMPs）和血管内皮生长因子-A（VEGF），这是一种非常有效的促血管生成因子。 后一个发现的含义是，虽然OSM可能导致体外乳腺癌细胞的生长停滞，但它也可能通过产生VEGF诱导肿瘤中的血管生成。 这种血管生成的诱导将通过促进血管生成依赖性乳腺癌进展来对抗OSM的生长停滞特性。 根据我们的研究结果，更好地理解VEGF诱导的意义以正确评估OSM作为临床癌症治疗的潜力是重要的。 该提案的具体目标是： 具体目标1. 证明嗜中性粒细胞衍生的OSM将以旁分泌方式从乳腺癌细胞诱导VEGF。 当中性粒细胞与乳腺癌细胞共培养时，它们会分泌OSM，但其病理生理意义尚不清楚。 将测试嗜中性粒细胞-乳腺癌细胞共培养物以确定嗜中性粒细胞分泌的OSM是否将以旁分泌方式从乳腺癌细胞诱导VEGF。将通过ELISA分析OSM和VEGF水平。 具体目标2. 识别和表征刺激中性粒细胞释放OSM的信号。 我们将通过对用特异性细胞因子（如GM-CSF）的中和抗体处理的共培养细胞的细胞裂解物/条件培养基进行蛋白质印迹/ELISA，鉴定乳腺癌细胞产生的刺激中性粒细胞释放OSM的信号。 此外，我们将确定嗜中性粒细胞-乳腺癌细胞接触是否需要在共培养系统（细胞接触）和Transwell培养（无细胞接触）中通过嗜中性粒细胞释放OSM。 具体目标3. 增强OSM诱导的VEGF在体内刺激血管生成和促进乳腺癌进展的能力。 我们将研究是否OSM诱导的VEGF将刺激血管生成在体内使用基质胶塞试验，并评估是否OSM转染的乳腺癌细胞注射到裸鼠的清除乳腺脂肪垫将促进肿瘤的进展，通过刺激VEGF依赖的血管生成的自分泌方式。