Invasive Behavior of Tumor Cells Producing Collagenase-1

产生胶原酶 1 的肿瘤细胞的侵袭行为

• 批准号: 8403840
• 负责人: CONSTANCE E BRINCKERHOFF
• 金额: $ 22.16万
• 依托单位: DARTMOUTH COLLEGE
• 依托单位国家: 美国
• 财政年份: 1998
• 资助国家: 美国
• 起止时间: 1998-12-15 至 2014-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8403840
• 关键词: Address; Affect; Autocrine Communication; Basement membrane; Behavior; Benign; Biology; Cells; Cessation of life; Chemicals; Clinical; Collagen Type I; Collagen Type IV; Data; Dermal; Dermis; Diagnosis; Endothelial Cells; Environment; Enzymes; Epidermis; Extracellular Matrix; Family; Frequencies; G-Protein-Coupled Receptors; Gelatinase A; Gelatinase B; Gene Expression; Growth; Health; Histologic; Human; Incidence; Interstitial Collagenase; Link; Lung; Malignant Neoplasms; Matrix Metalloproteinases; Mediating; Melanoma Cell; Metastatic Lesion; Metastatic Melanoma; Metastatic to; Morbidity - disease rate; Neoplasm Metastasis; Neutrophil Collagenase; Nude Mice; Outcome; PAR-1 Receptor; Paracrine Communication; Pathway interactions; Patients; Phase Transition; Phenotype; Primary Neoplasm; Process; Production; Prognostic Marker; RNA Interference; Radial Growth Phase; Resistance; Role; Serine Protease; Signal Transduction; Signal Transduction Pathway; Site; Skin Cancer; Staging; Technology; Testing; Thrombin; Transcription Factor AP-2 Alpha; Tumor Cell Invasion; Vertical Growth Phase; angiogenesis; autocrine; base; cell behavior; collagenase; collagenase 1; collagenase 3; human MMP14 protein; in vivo; inhibitor/antagonist; interstitial; knock-down; melanoma; mortality; neoplastic cell; neutralizing antibody; new therapeutic target; novel; novel strategies; paracrine; prevent; small hairpin RNA; transcription factor; tumor; tumor growth; tumor progression; Address; Affect; Autocrine Communication; Basement membrane; Behavior; Benign; Biology; Cells; Cessation of life; Chemicals; Clinical; Collagen Type I; Collagen Type IV; Data; Dermal; Dermis; Diagnosis; Endothelial Cells; Environment; Enzymes; Epidermis; Extracellular Matrix; Family; Frequencies; G-Protein-Coupled Receptors; Gelatinase A; Gelatinase B; Gene Expression; Growth; Health; Histologic; Human; Incidence; Interstitial Collagenase; Link; Lung; Malignant Neoplasms; Matrix Metalloproteinases; Mediating; Melanoma Cell; Metastatic Lesion; Metastatic Melanoma; Metastatic to; Morbidity - disease rate; Neoplasm Metastasis; Neutrophil Collagenase; Nude Mice; Outcome; PAR-1 Receptor; Paracrine Communication; Pathway interactions; Patients; Phase Transition; Phenotype; Primary Neoplasm; Process; Production; Prognostic Marker; RNA Interference; Radial Growth Phase; Resistance; Role; Serine Protease; Signal Transduction; Signal Transduction Pathway; Site; Skin Cancer; Staging; Technology; Testing; Thrombin; Transcription Factor AP-2 Alpha; Tumor Cell Invasion; Vertical Growth Phase; angiogenesis; autocrine; base; cell behavior; collagenase; collagenase 1; collagenase 3; human MMP14 protein; in vivo; inhibitor/antagonist; interstitial; knock-down; melanoma; mortality; neoplastic cell; neutralizing antibody; new therapeutic target; novel; novel strategies; paracrine; prevent; small hairpin RNA; transcription factor; tumor; tumor growth; tumor progression

DESCRIPTION (provided by applicant): Destruction of the extracellular matrix in tumor invasion and metastasis is mediated primarily by Matrix Metalloproteinases (MMPs), a family of enzymes that are over-expressed in many cancers. Degradation of type IV collagen in basement membrane by MMP-2 and MMP-9 is an essential step, but degradation of the interstitial collagens (types I and III) is also required. This is accomplished by the collagenases, of which MMP-1 is the most ubiquitously expressed. Malignant melanoma is an aggressive cancer where MMP-1 contributes to an invasive phenotype and is associated with poor outcome. Melanoma is the most common fatal skin cancer, with approximately 59,000 new cases diagnosed and 8,000 patient deaths in 2007. Melanoma progression from benign to metastatic tumor is classified histologically, and early stage melanoma is defined as radial growth phase (RGP), which is confined to the epidermis. In contrast, later stage vertical growth phase (VGP) is characterized by invasion of melanoma cells into the dermis, and VGP frequently progresses to metastatic melanoma. However, mechanisms mediating the RGP to VGP transition are not known. Increasing evidence points to Protease Activated Receptor-1 (PAR-1), a G- coupled protein receptor, as critical in the progression to VGP. Based on our Preliminary Data, we hypothesize that an autocrine MMP-1/PAR-1 axis in RGP melanoma initiates signaling cascades that facilitate the RGP to VGP transition, and that this autocrine signaling axis then contributes to the metastatic behavior of VGP melanomas. We also hypothesize a paracrine signaling axis in which melanoma-derived MMP-1 activates PAR-1 on endothelial cells to mediate angiogenesis and melanoma progression. To determine how MMP-1 contributes to melanoma progression, we inhibited MMP-1 production in VGP melanoma cells with RNA interference (RNAi) technology. Cells stably expressing MMP-1 shRNAs had > 85% knock-down of MMP-1 compared to MAMMX control shRNAs, and when injected interdermally into nude mice, MMP-1 shRNAs and shMAMMX cells formed primary tumors with similar growth rates. However, shMAMMX tumors metastasized to the lung, while tumors from MMP-1 shRNA cells did not. Further, melanoma-produced MMP-1 mediated invasion through the dermis, and the MMP-/PAR-1 paracrine signaling axis enhanced angiogenesis. Thus, based on these data, we now hypothesize that tumor-produced MMP-1 mediates melanoma growth and metastasis via novel autocrine and paracrine activation of PAR-1 signaling, initiation of pathways transduction pathways and changes in gene expression. We will test this hypothesis by investigating: (1) the autocrine MMP-1/PAR-1 signaling axis in the RGP to VGP transition and in the metastatic behavior of VGP melanoma; (2) the MMP-1/PAR-1 paracrine axis in endothelial cell behavior; and (3) using siRNAs, the mechanisms by which MMP-1 facilitates tumor growth at metastatic sites. Our studies will define novel roles for MMP-1 in melanoma progression.

描述（由申请人提供）：肿瘤侵袭和转移中细胞外基质的破坏主要是由基质金属蛋白酶（MMP）介导的，金属蛋白酶（MMP）是一个在许多癌症中过表达的酶家族。 MMP-2和MMP-9在基底膜中IV型胶原蛋白的降解是必不可少的步骤，但是还需要间隙胶原蛋白（I型和III）的降解。这是通过胶原酶完成的，其中MMP-1是最普遍的表达。恶性黑色素瘤是一种侵略性癌症，其中MMP-1有助于侵袭性表型，并且与不良预后有关。黑色素瘤是最常见的致命皮肤癌，2007年约有59,000例诊断出的新病例和8,000例患者死亡。黑色素瘤从良性到转移性肿瘤的进展被组织学分类，早期黑色素瘤被定义为径向生长期（RGP），仅限于表皮。相反，后期垂直生长阶段（VGP）的特征是黑色素瘤细胞进入真皮，而VGP经常发展为转移性黑色素瘤。但是，介导RGP到VGP过渡的机制尚不清楚。增加的证据表明蛋白酶活化受体-1（PAR-1）是一种G偶联蛋白受体，这对于向VGP的发展至关重要。根据我们的初步数据，我们假设RGP黑色素瘤中的自分泌MMP-1/PAR-1轴启动了信号级联，从而促进RGP到VGP过渡，然后该自动分泌信号轴，然后有助于VGP Melanomas的转移性行为。我们还假设一个旁分泌信号轴，其中黑色素瘤衍生的MMP-1激活内皮细胞上的PAR-1以介导血管生成和黑色素瘤进展。为了确定MMP-1如何促进黑色素瘤进展，我们用RNA干扰（RNAI）技术抑制了VGP黑色素瘤细胞中的MMP-1产生。与MAMMX对照shRNA相比，稳定表达MMP-1 shRNA的细胞的MMP-1击倒> 85％，并且当将衍生品注射到裸鼠中时，MMP-1 SHRNAS和SHMAMMX细胞形成了具有相似生长速率的原发性肿瘤。然而，SHMAMMX肿瘤转移到肺部，而MMP-1 shRNA细胞的肿瘤没有。此外，黑色素瘤产生的MMP-1通过真皮介导的浸润，MMP-/PAR-1旁分泌信号传导轴增强了血管生成。因此，基于这些数据，我们现在假设肿瘤产生的MMP-1通过新的自分泌和旁分泌激活PAR-1信号传导，途径转导途径的启动以及基因表达的变化来介导黑色素瘤的生长和转移。我们将通过研究：（1）RGP到VGP过渡中的自分泌MMP-1/PAR-1信号轴以及VGP黑色素瘤的转移行为； （2）内皮细胞行为中的MMP-1/PAR-1旁分泌轴； （3）使用siRNA，MMP-1促进转移性部位肿瘤生长的机制。我们的研究将定义MMP-1在黑色素瘤进展中的新作用。