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.

人嗅蛋白4基因（OLFM4）编码嗅觉素相关的糖蛋白。 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在很大程度上未知。在这项研究中，我们发现在前列腺癌组织和前列腺癌细胞系中降低或无法检测到OLFM4信使RNA。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，发现表达OLFM4的PC-3细胞克隆的增殖和侵入性降低。此外，与注射载体转染的对照细胞相比，注射了表达OLFM4的PC-3细胞的裸鼠表现出肿瘤的生长，骨骼侵袭和转移的降低。机械研究表明，OLFM4通过靶向组织蛋白酶D来调节细胞自噬可能表现出其抗癌作用，因为OLFM4降低了组织蛋白酶D蛋白D蛋白水平和酶活性，并减弱了组织蛋白酶D诱导的癌细胞增殖。另外，OLFM4的过表达废除了基质细胞衍生的因子1（SDF-1）诱导的PC-3细胞侵入性，部分通过阻断SDF-1介导的AKT磷酸化，部分地通过阻断了SDF-1介导的AKT磷酸化。在表达OLFM4的PC-3细胞中的共免疫沉淀和免疫荧光染色研究表明OLFM4和组织蛋白酶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负相互作用，并抑制前列腺癌的生长和骨转移。为了进一步研究OLFM4基因在前列腺组织中的生理功能，我们已经开发并分析了OLFM4敲除小鼠模型。在去年，我们发现OLFM4损失与临床样品中的前列腺癌进展有关，并在小鼠模型中。在患者标本中，我们发现使用激光捕获微分辨率在癌性前列腺上皮细胞中OLFM4外显子2中2298231（S118S）的杂合性损失。我们进一步通过荧光原位杂交分析发现了前列腺癌样品中OLFM4基因的半合子和纯合缺失。有趣的是，OLFM4基因敲除小鼠在1824个月的年龄在1824个月的前列腺内肿瘤（44％）和前列腺腺癌（28％）出现，表达微阵列分析表明，这些前列腺样品中与细胞增殖相关的基因水平增加。特别是，OLFM4的丧失产生了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，我们表明OLFM4-敲除小鼠偶发发展的前列腺上皮内肿瘤（44％）（44％），前列腺腺癌（28％）和其他1824个月大的肿瘤。来自OLFM4缺陷小鼠的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蛋白通过与声音刺猬蛋白直接相互作用，抑制了前列腺茎/祖细胞增殖和介导的刺猬信号通道活动。综上所述，我们的数据表明OLFM4在前列腺肿瘤进程中起肿瘤抑制剂的作用。