Olfactomedin 4 Suppresses Prostate Cancer Cell Growth and Metastasis via Negativ

Olfactomedin 4 通过 Negativ 抑制前列腺癌细胞的生长和转移

基本信息

批准号：
8746603
负责人：
GRIFFIN RODGERS
金额：
$ 38.54万
依托单位：
NATIONAL HEART, LUNG, AND BLOOD INSTITUTE
依托单位国家：
美国
项目类别：
财政年份：
资助国家：
美国
起止时间：
至
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/8746603
关键词：
13q12-13 13q14 13q14.3 14q Age Age-Months American Attenuated Autophagocytosis BRCA2 gene Biological Biological Process CXCL12 gene Cadherins Cancer Cell Growth Cancer Etiology Cancerous Cathepsins Cell Adhesion Cell Proliferation Cell surface Cells Cessation of life Chromosomes Clinical Clone Cells Colon Carcinoma Colonic Neoplasms Data Diagnosis ETS Family Gene Ectopic Expression Epithelial Cells Evaluation Exhibits Exons Family Family member Fluorescent in Situ Hybridization Gene Expression Gene Proteins Genes Glycoproteins Goals Growth Human Human Genome Immunofluorescence Immunologic In Vitro Inflammation Knockout Mice Lectin Loss of Heterozygosity Malignant Neoplasms Malignant neoplasm of prostate Matrigel Invasion Assay Mediating Messenger RNA Metastatic Neoplasm to the Bone Microarray Analysis Molecular Molecular Profiling Mus Natural Immunity Neoplasm Metastasis Nude Mice PC3 cell line Patients Phosphorylation Physiological Play Prostate Prostate Adenocarcinoma Prostatic Intraepithelial Neoplasias Prostatic Neoplasms Proteins Proto-Oncogene Proteins c-akt RB1 gene Reporting Retinoblastoma Role Sampling Seminal fluid Serum Solid Neoplasm Sonic hedgehog protein Specimen Staining method Stains Stem cells Stomach Neoplasms Stromal Cell-Derived Factor 1 Susceptibility Gene Tissues Tumor Suppressor Genes Tumor Suppressor Proteins bone invasion cancer cell cancer type forkhead protein human GW112 protein improved in vivo laser capture microdissection malignant breast neoplasm malignant stomach neoplasm matrix metalloproteinase 19 men mouse model olfactomedin outcome forecast overexpression prostate cancer cell smoothened signaling pathway stem tumor tumor growth tumor progression vector

项目摘要

The human olfactomedin 4 gene (OLFM4) encodes an olfactomedin-related glycoprotein. OLFM4 is normally expressed in a limited number of tissues, including the prostate, but its biological functions in prostate are largely unknown. In this study, we found that OLFM4 messenger RNA was reduced or undetectable in prostate cancer tissues and prostate cancer cell lines. To study the effects of OLFM4 on prostate cancer progression, we transfected PC-3 prostate cancer cells with OLFM4 to establish OLFM4-expressing PC-3 cell clones. The OLFM4-expressing PC-3 cell clones were found to have decreased proliferation and invasiveness compared with vector-transfected control PC-3 cells in vitro. In addition, nude mice injected with OLFM4-expressing PC-3 cells demonstrated reduced tumor growth and bone invasion and metastasis compared with mice injected with vector-transfected control cells. Mechanistic studies revealed that OLFM4 may exhibit its anticancer effects through regulating cell autophagy by targeting cathepsin D, as OLFM4 reduced cathepsin D protein levels and enzymatic activity and attenuated cathepsin D-induced cancer cell proliferation. In addition, overexpression of OLFM4 abrogated stromal cell derived factor-1 (SDF-1)-induced PC-3 cell invasiveness in a Matrigel invasion assay, partially through blocking SDF-1-mediated AKT phosphorylation. Coimmunoprecipitation and immunofluorescence staining studies in OLFM4-expressing PC-3 cells demonstrated a direct interaction between OLFM4 and cathepsin D or SDF-1. Taken together, these results suggest that OLFM4 negatively interacts with cathepsin D and SDF-1 and inhibits prostate cancer growth and bone metastasis. To further investigate the physiological functions of OLFM4 gene in the prostate tissue, we have developed and analyzed an OLFM4 knock out mouse model.During this last year, we found that OLFM4 loss is associated with prostate cancer progression in clinical samples and in a mouse model. In patient specimens, we found loss of heterozygosity at rs 2298231(S118S) within exon 2 of OLFM4 in cancerous prostate epithelial cells using laser-capture microdissection. We further found hemizygous and homozygous deletions of the OLFM4 gene in prostate cancer samples via fluorescence in situ hybridization analysis. Interestingly, OLFM4 knockout mice developed prostatic intraepithelial neoplasia (44%) and prostatic adenocarcinoma (28%) at ages 1824 months, and expression microarray analysis demonstrated increased levels of genes associated with cell proliferation in these prostate samples. In particular, loss of OLFM4 produced increased expression of the ETS family gene Ets1 and the cancer-promoting matrix metalloproteinase 19 (MMP19). In our most recent studies, we show that Olfm4-knockout mice sporadically developed prostatic intraepithelial neoplasia (44%), prostatic adenocarcinoma (28%), and other tumors at 1824 months of age. The gene-expression profile of prostate tissues from Olfm4-deficient mice revealed significant changes for genes associated with neoplastic progression. Human OLFM4 protein inhibited prostate stem/progenitor cell proliferation and mediated hedgehog signaling-pathway activities through direct interaction with sonic hedgehog protein. Taken together, our data indicate that OLFM4 plays a tumor-suppressor role in prostate neoplastic progression.

人类嗅觉定 4 基因 (OLFM4) 编码嗅觉定相关糖蛋白。 OLFM4通常在包括前列腺在内的有限数量的组织中表达，但其在前列腺中的生物学功能很大程度上未知。在这项研究中，我们发现 OLFM4 信使 RNA 在前列腺癌组织和前列腺癌细胞系中减少或检测不到。为了研究 OLFM4 对前列腺癌进展的影响，我们用 OLFM4 转染 PC-3 前列腺癌细胞以建立表达 OLFM4 的 PC-3 细胞克隆。与载体转染的对照 PC-3 细胞相比，在体外发现表达 OLFM4 的 PC-3 细胞克隆的增殖和侵袭性降低。此外，与注射载体转染的对照细胞的小鼠相比，注射表达OLFM4的PC-3细胞的裸鼠表现出肿瘤生长以及骨侵袭和转移减少。机制研究表明，OLFM4 可能通过靶向组织蛋白酶 D 来调节细胞自噬，从而发挥抗癌作用，因为 OLFM4 降低组织蛋白酶 D 蛋白水平和酶活性，并减弱组织蛋白酶 D 诱导的癌细胞增殖。此外，在 Matrigel 侵袭试验中，OLFM4 的过表达消除了基质细胞衍生因子 1 (SDF-1) 诱导的 PC-3 细胞侵袭性，部分是通过阻断 SDF-1 介导的 AKT 磷酸化来实现的。在表达 OLFM4 的 PC-3 细胞中进行的免疫共沉淀和免疫荧光染色研究表明 OLFM4 与组织蛋白酶 D 或 SDF-1 之间存在直接相互作用。综上所述，这些结果表明 OLFM4 与组织蛋白酶 D 和 SDF-1 产生负相互作用，并抑制前列腺癌生长和骨转移。为了进一步研究 OLFM4 基因在前列腺组织中的生理功能，我们开发并分析了 OLFM4 敲除小鼠模型。去年，我们在临床样本和小鼠模型中发现 OLFM4 缺失与前列腺癌进展相关。在患者样本中，我们使用激光捕获显微切割发现癌性前列腺上皮细胞中 OLFM4 外显子 2 内的 rs 2298231(S118S) 杂合性丢失。我们通过荧光原位杂交分析进一步发现前列腺癌样本中 OLFM4 基因的半合子和纯合子缺失。有趣的是，OLFM4 基因敲除小鼠在 1824 个月时出现前列腺上皮内瘤变 (44%) 和前列腺腺癌 (28%)，表达微阵列分析表明这些前列腺样本中与细胞增殖相关的基因水平增加。特别是，OLFM4 的缺失导致 ETS 家族基因 Ets1 和促癌基质金属蛋白酶 19 (MMP19) 的表达增加。在我们最近的研究中，我们发现 Olfm4 敲除小鼠在 1824 月龄时偶发性出现前列腺上皮内瘤变 (44%)、前列腺腺癌 (28%) 和其他肿瘤。 Olfm4 缺陷小鼠前列腺组织的基因表达谱揭示了与肿瘤进展相关的基因的显着变化。人 OLFM4 蛋白抑制前列腺干/祖细胞增殖，并通过与 sonic hump 蛋白直接相互作用介导 hedgehog 信号通路活性。综上所述，我们的数据表明 OLFM4 在前列腺肿瘤进展中发挥肿瘤抑制作用。