title: High throughput quantitative reverse transcription PCR assays revealing over-expression of cancer testis antigen genes in multiple myeloma stem cell-like side population cells.
abstract: Multiple myeloma (MM) stem cells, proposed to be responsible for the tumourigenesis, drug resistance and recurrence of this disease, are enriched in the cancer stem cell-like side population (SP). Cancer testis antigens (CTA) are attractive targets for immunotherapy because they are widely expressed in cancers but only in limited types of normal tissues. We designed a high throughput assay, which allowed simultaneous relative quantifying expression of 90 CTA genes associated with MM. In the three MM cell lines tested, six CTA genes were over-expressed in two and LUZP4 and ODF1 were universally up-regulated in all three cell lines. Subsequent study of primary bone marrow (BM) from eight MM patients and four healthy donors revealed that 19 CTA genes were up-regulated in SP of MM compared with mature plasma cells. In contrast, only two CTA genes showed a moderate increase in SP cells of healthy BM. Furthermore, knockdown using small interfering RNA (siRNA) revealed that LUZP4 expression is required for colony-forming ability and drug resistance in MM cells. Our findings indicate that multiple CTA have unique expression profiles in MM SP, suggesting that CTA may serve as targets for immunotherapy that it specific for MM stem cells and which may lead to the long-term cure of MM.
authors.affiliation: Department of Pathology and Genomic Medicine, Houston Methodist Hospital, Houston, TX, USA.
authors.forename: Jianguo
authors.initials: J
authors.lastname: Wen
authors.forename: Hangwen
authors.initials: H
authors.lastname: Li
authors.forename: Wenjing
authors.initials: W
authors.lastname: Tao
authors.forename: Barbara
authors.initials: B
authors.lastname: Savoldo
authors.forename: Jessica A
authors.initials: JA
authors.lastname: Foglesong
authors.forename: Lauren C
authors.initials: LC
authors.lastname: King
authors.forename: Youli
authors.initials: Y
authors.lastname: Zu
authors.forename: Chung-Che
authors.initials: CC
authors.lastname: Chang
mesh_terms: D000951:Antigens, Neoplasm / Q000032:analysis* / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology; D017209:Apoptosis / Q000276:immunology; D014408:Biomarkers, Tumor / Q000032:analysis* / Q000096:biosynthesis / Q000235:genetics; D045744:Cell Line, Tumor; D004268:DNA-Binding Proteins / Q000032:analysis / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology; D006360:Heat-Shock Proteins / Q000032:analysis / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology; D057166:High-Throughput Screening Assays / Q000379:methods; D006801:Humans; D009101:Multiple Myeloma / Q000235:genetics* / Q000276:immunology / Q000473:pathology*; D014411:Neoplastic Stem Cells / Q000276:immunology*; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods
chemical_list: D000951:Antigens, Neoplasm; D014408:Biomarkers, Tumor; D004268:DNA-Binding Proteins; D006360:Heat-Shock Proteins; C456621:LUZP4 protein, human; C086058:ODF1 protein, human
keywords: cancer testis antigen; high throughput; multiple myeloma; quantitative RT-PCR; side population