GLYCOSPHINGOLIPID ENRICHED MICRODOMAINS IN CANCER CELL INVASION

鞘糖脂富集癌细胞侵袭中的微结构域

• 批准号: 7720455
• 负责人: WIM Floris Albert STEELANT
• 金额: $ 12.62万
• 依托单位: NEW MEXICO STATE UNIVERSITY LAS CRUCES
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-05-01 至 2009-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7720455
• 关键词: Adhesions; Antigens; Behavior; Breast; Breast Cancer Cell; CA-15-3 Antigen; Cell Adhesion; Cell Adhesion Molecules; Cell membrane; Cell surface; Cell-Cell Adhesion; Cells; Clinical; Collaborations; Colon Carcinoma; Complex; Computer Retrieval of Information on Scientific Projects Database; Data; E-Cadherin; Endopeptidases; Ethers; Ethyl Ether; Event; Experimental Models; Funding; Future; Glycosphingolipids; Goals; Grant; Human; Institution; Integrin alpha1; Integrins; Invasive; LNCaP; Laboratory Animal Production and Facilities; Lead; Link; Lipids; Localized; Location; MCF7 cell; Malignant Neoplasms; Malignant neoplasm of prostate; Matrix Metalloproteinases; Mediating; Mediator of activation protein; Membrane Microdomains; Membrane Proteins; Metastatic Prostate Cancer; Modeling; Molecular Probes; Mus; New Mexico; PTK2 gene; Pathway interactions; Peptide Hydrolases; Phenotype; Phosphorylcholine; Play; Process; Prostate; Proteins; Research; Research Personnel; Resources; Role; Signal Pathway; Signal Transduction; Signaling Molecule; Source; Specimen; Transducers; Tumor Cell Invasion; United States National Institutes of Health; Universities; Validation; Work; Xenograft Model; analog; cancer cell; clinically relevant; human BCAR1 protein; in vivo; medical schools; sugar; 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. Glycosphingolipid Enriched Microdomains in Cancer Cell Invasion The ultimate goal for understanding and combating cancer progression is to investigate mechanisms culminating in an invasive and metastatic cell phenotype. Cancer cell invasion is a highly complex and multistep process, that is characterized by altered expression levels of cell adhesion molecules and secretion of proteolytic enzymes, along with changes in expression or activities of a variety of cellular proteins in multiple branching signaling pathways. In addition, the plasma membrane contains membrane microdomains enriched in tumor-associated glycosphingolipid (GSL)-antigens that function as mediators of cell adhesion, and in signaling molecules, providing important locations for cell signaling. Recent studies showed that tumor cell invasion often results from aberrant signaling mechanisms initiated by differential organization and clustering of GSLs and/or membrane proteins, and their assembly with signaling molecules. The synthetic ether lipid analog ET-18-OMe (1-O-octadecyl-2-O-methyl-glycero-3-phosphocholine) was previously shown to influence invasion of human breast and colon cancer cells and is used as a molecular probe to induce cellular invasion in order to explore changes in composition, translocation and organization of crucial molecules and the possible involvement of membrane microdomains. Our results show that reorganization, clustering and assembly of specific membrane proteins and signal transducers with glycosphingolipids in membrane microdomains are responsible for loss of cell-cell adhesion and subsequent result in increased invasiveness of MCF-7 breast cancer cells. Where loss of cell-cell adhesion was due to sterical hindrance of E-cadherin by episialin both localized in and associated with clusters of the glycosphingolipid, MSGb5, subsequent invasion resulted from activation of MSGb5-associated FAK/cSrc signaling complexes and downstream ERK, leading to increased expression and activation of MMPs. In contrast, the invasive behavior of HCT-8 colon cancer cells was initiated through clustering of integrin alpha1 subunits and the activation of associated FAK/cSrc complexes. The followed path downstream, however, appeared to be the p130Cas-JNK pathway also leading to increased expression of MMPs and did not involve specific association with glycosphingolipids. The latter results point out to the 'classic' integrin-dependent invasion of cancer cells, initiated through clustering of integrins. In contrast, signaling towards invasion may also occur via GSLs, adhesion to the surrounding matrix mediated by the sugar moieties of glycosphingolipids resulting in GSL-dependent adhesion may then induce signaling events to alter cellular phenotype. These results suggested that cancer malignancy may result from reorganization of existing components that lead to changes in intracellular signaling events. Therefore, we choose to continue our study with the LNCaP/C4-2/C4-2B progression model, which closely resembles the clinical progression of human prostate cancer. This experimental model enables the study of mechanistic alterations of metastatic prostate cancer cells compared to their parental cells. Our preliminary data indicate that FAK and src, that play a crucial role in invasion, are activated in the metastatic subline C4-2B, and are involved in enhanced expression and activity of matrix metalloproteinases. In addition, alterations in cell surface organization of possible upstream regulators that may trigger these cellular events were demonstrated and are currently investigated to further elucidate the relevance of their organization which may link them to activation of downstream signaling pathways and thus alteration of phenotype. Future work includes assessment in clinically relevant specimen and validation of our findings in vivo using xenograft models in mice in collaboration with Dr. Marco Bisoffi from the University of New Mexico, School of Medicine at the UNM Animal Research Facility.

该副本是利用众多研究子项目之一 由NIH/NCRR资助的中心赠款提供的资源。子弹和 调查员（PI）可能已经从其他NIH来源获得了主要资金， 因此可以在其他清晰的条目中代表。列出的机构是 对于中心，这不一定是调查员的机构。 癌细胞浸润中富集糖脂的微域 理解和打击癌症进展的最终目标是研究最终在侵入性和转移细胞表型中的机制。癌细胞侵袭是一种高度复杂且多步的过程，其特征是细胞粘附分子的表达水平改变和蛋白水解酶的分泌，以及多种分支信号途径中多种细胞蛋白的表达或活性的变化。此外，质膜含有富含肿瘤相关的糖磷脂（GSL）抗原的膜微域，可作为细胞粘附的介体和信号分子中，为细胞信号传导提供重要位置。最近的研究表明，肿瘤细胞的侵袭通常是由于差异组织和GSL和/或膜蛋白的聚类引发的异常信号传导机制以及它们与信号分子的组装。 The synthetic ether lipid analog ET-18-OMe (1-O-octadecyl-2-O-methyl-glycero-3-phosphocholine) was previously shown to influence invasion of human breast and colon cancer cells and is used as a molecular probe to induce cellular invasion in order to explore changes in composition, translocation and organization of crucial molecules and the possible involvement of membrane microdomains. 我们的结果表明，在膜微区中，特定膜蛋白和信号传感器的重组，聚类和组装和信号传感器在膜微区内的糖果脂质是导致细胞细胞粘附的丧失，随后导致MCF-7乳腺癌细胞的侵袭性增加。 eScialin均因糖化蛋白对E-钙粘蛋白的静脉障碍而导致的细胞细胞粘附丧失，并且与糖磷脂的簇相关，MSGB5随后侵袭MSGB5相关的FAK/CSRC/CSRC信号传导复合物和下游ERK，导致表达和激活的表达和Activation MMMPS，引起了MSGB5相关的FAK/CSRC信号复合物。相反，HCT-8结肠癌细胞的侵入性行为是通过整合素α1亚基和相关FAK/CSRC复合物的激活而启动的。然而，下游的跟踪路径似乎是P130CAS-JNK途径，也导致MMP的表达增加，并且不涉及与糖磷脂的特定关联。后者的结果指出了通过整合素聚类引发的癌细胞的“经典”整合素依赖性侵袭。相比之下，也可能通过GSL进行侵袭的信号传导，对周围基质的粘附是由糖磷脂脂的糖部分介导的，从而导致GSL依赖性粘附，然后可能引起信号事件以改变细胞表型。这些结果表明，癌症恶性肿瘤可能是由于现有组件的重组而导致细胞内信号事件发生变化的原因。因此，我们选择使用LNCAP/C4-2/C4-2B进展模型继续研究，该模型与人前列腺癌的临床进展非常相似。该实验模型可以研究与亲本细胞相比，将转移性前列腺癌细胞的机械改变。我们的初步数据表明，FAK和SRC在侵袭中起着至关重要的作用，在转移性subline C4-2B中被激活，并且参与了基质金属蛋白酶的增强表达和活性。此外，证明了可能触发这些细胞事件的可能上游调节剂细胞表面组织的变化，目前进行了研究，以进一步阐明其组织的相关性，这可能将它们与下游信号通路的激活联系起来，从而改变了表型。未来的工作包括对临床相关标本的评估以及与新墨西哥大学Marco Bisoffi博士合作，使用小鼠中的异种移植模型对我们的体内发现的验证，UNM动物研究机构的医学院。