EPIGENETIC EFFECTS OF CERAMIDE GLYCOSYLATION AND DRUG-RESISTANT CANCER STEM CELL

神经酰胺糖基化与耐药癌症干细胞的表观遗传效应

• 批准号: 8360364
• 负责人: Yong-Yu Liu
• 金额: $ 16.48万
• 依托单位: LOUISIANA STATE UNIV A&M COL BATON ROUGE
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-05-01 至 2012-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8360364
• 关键词: Antineoplastic Agents; Biomedical Research; CD44 gene; Cell Line; Cells; Ceramide glucosyltransferase; Ceramides; Doxorubicin; Drug resistance; Epigenetic Process; FGF1 gene; Funding; Genes; Glycosphingolipids; Goals; Grant; Human; Louisiana; MCF7 cell; Mediating; Mixed-Backbone Oligonucleotide; National Center for Research Resources; Neoplasm Metastasis; Nude Mice; Phenotype; Principal Investigator; Research; Research Infrastructure; Resistance; Resources; Role; Series; Sorting - Cell Movement; Source; Tumorigenicity; United States National Institutes of Health; cancer cell; cancer stem cell; chemotherapy; cost; enzyme activity; glycosylation; in vivo; malignant breast neoplasm; overexpression; tumor; tumor growth; tumor progression

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. Anticancer drugs induce tumor drug-resistance and metastasis, and cancer stem cells display more resistant than other differentied cancer cells; however, little is known whether anticancer drugs promote cancer stem cells. The goal of this project is to determine the role of ceramide glycosylation in mediating cancer stem cells during the course of chemotherapy. To complet the goal, we pursued two specific aims in last year: 1), determine the increasements of glucosylceramide synthase (GCS) and breast cancer stem cell (BCSC) in drug-resistant cancer cells; 2), identify the induction of doxorubicin in cancer stem cells via ceramide glycosylation in vivo. We found that GCS overexpression was interrelated to the increment BCSCs and drug-resistance in human breast cancer MCF-7 cell lines after doxorubicin induction. With GCS overexpression, the BCSCs of CD24-/CD44+/ESA+ phenotype was increased by 5-fold in MCF-7/Dox cells as compared to MCF-7 cells. Silencing GCS by using mixed-backbone oligonucleotide (MBO-asGCS) significantly decreased the numbers of BCSCs more than 2-fold. BCSCs sorted displayed 3-fold higher GCS enzyme activity and formed 2-fold more colonies, as compare with other non-stem cell subsets. The BCSCs cells showed significantly higher tumorigenicity and metastasibility as compared to non-stem cells in athymic nude mice. More interestingly, doxorubicin treatment considerably increased BCSCs by 2-fold in tumors, and MBO-asGCS treatment eliminated the tumor growth and reduced metastasis due to reduction of BCSCs. Comparison of glycosphingolipids and gene profile indicated GCS or globo-series glycosphingolipids regulates FGF1 and others to accumulate BCSCs. These results demonstrate that ceramide glycosylation accumulates BCSCs formation and causes aggressive tumor progression.

该子项目是利用资源的众多研究子项目之一 由 NIH/NCRR 资助的中心拨款提供。子项目的主要支持 并且子项目的主要研究者可能是由其他来源提供的， 包括其他 NIH 来源。 子项目可能列出的总成本 代表子项目使用的中心基础设施的估计数量， NCRR 赠款不直接向子项目或子项目工作人员提供资金。 抗癌药物诱导肿瘤耐药和转移，癌症干细胞比其他分化癌细胞表现出更强的耐药性；然而，人们对抗癌药物是否促进癌症干细胞知之甚少。该项目的目标是确定神经酰胺糖基化在化疗过程中介导癌症干细胞中的作用。为了完成这一目标，我们去年追求了两个具体目标：1）确定耐药癌细胞中葡萄糖神经酰胺合酶（GCS）和乳腺癌干细胞（BCSC）的增加； 2)、确定体内通过神经酰胺糖基化在癌症干细胞中诱导阿霉素。我们发现，在阿霉素诱导后，GCS 过表达与人乳腺癌 MCF-7 细胞系中 BCSC 的增加和耐药性相关。随着 GCS 过表达，与 MCF-7 细胞相比，MCF-7/Dox 细胞中 CD24-/CD44+/ESA+ 表型的 BCSC 增加了 5 倍。使用混合主链寡核苷酸 (MBO-asGCS) 沉默 GCS 可使 BCSC 数量显着减少 2 倍以上。与其他非干细胞亚群相比，分选的 BCSC 显示出 3 倍高的 GCS 酶活性，并形成了 2 倍多的集落。与无胸腺裸鼠中的非干细胞相比，BCSCs细胞表现出显着更高的致瘤性和转移性。更有趣的是，阿霉素治疗使肿瘤中的 BCSC 显着增加了 2 倍，而 MBO-asGCS 治疗则消除了肿瘤生长，并由于 BCSC 减少而减少了转移。鞘糖脂和基因谱的比较表明GCS或globo系列鞘糖脂调节FGF1和其他物质以积累BCSC。这些结果表明神经酰胺糖基化会累积 BCSC 形成并导致侵袭性肿瘤进展。