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.

该副本是利用资源的众多研究子项目之一 由NIH/NCRR资助的中心赠款提供。对该子弹的主要支持 而且，副投影的主要研究员可能是其他来源提供的 包括其他NIH来源。 列出的总费用可能 代表subproject使用的中心基础架构的估计量， NCRR赠款不直接向子弹或副本人员提供的直接资金。 抗癌药物诱导肿瘤药物抗性和转移，癌症干细胞比其他不同的癌细胞表现出更具耐药性。但是，鲜为人知的抗癌药是否促进癌症干细胞。该项目的目的是确定神经酰胺糖基化在化学疗法过程中介导癌症干细胞中的作用。为了完成目标，我们在去年实现了两个具体目标：1），确定葡萄糖基酶合酶（GCS）和乳腺癌干细胞（BCSC）在药物耐药性癌细胞中的增加； 2），确定通过体内神经酰胺糖基化在癌症干细胞中诱导阿霉素的诱导。我们发现，在阿霉素诱导后，GCS过表达与人乳腺癌MCF-7细胞系的增量BCSC和药物抗性相关。使用GCS过表达，与MCF-7细胞相比，在MCF-7/DOX细胞中，CD24-/CD44+/ESA+表型的BCSC增加了5倍。通过使用混合背骨寡核苷酸（MBO-ASGC）沉默的GCS可显着减少BCSC的数量超过2倍。与其他非茎细胞亚群相比，BCSC分类的BCSC表现出3倍高的GCS酶活性，并形成2倍的菌落。与无胸腺裸鼠的非茎细胞相比，BCSC细胞显示出明显更高的致瘤性和转移性。更有趣的是，阿霉素治疗在肿瘤中大大增加了BCSC的2倍，而MBO-ASGCS治疗消除了由于BCSC的减少而消除了肿瘤的生长和降低的转移。糖磷脂和基因谱的比较表明GCS或Globo系列糖果脂脂调节FGF1和其他人积累BCSC。这些结果表明，神经酰胺糖基化积累了BCSC的形成并引起攻击性肿瘤进展。