title: Breast cancer cells expressing stem cell markers CD44+ CD24 lo are eliminated by Numb-1 peptide-activated T cells.
abstract: Cancer stem cells (CSC) are resistant to chemo- and radiotherapy. To eliminate cells with phenotypic markers of CSC-like we characterized: (1) expression of CD44, CD24, CD133 and MIC-A/B (NKG2 receptors) in breast (MCF7) and ovarian (SK-OV-3) cells resistant to gemcitabine (GEM), paclitaxel (PTX) and 5-fluorouracil (5-FU) and (2) their elimination by Numb- and Notch-peptide activated CTL. The number of cells in all populations with the luminal CSC phenotype [epithelial specific antigen(+) (ESA) CD44(hi) CD24(lo), CD44(hi) CD133(+), and CD133(+) CD24(lo)] increased in drug-resistant MCF7 and SK-OV-3 cells. Similarly, the number of cells with expressed MIC-A/B increased 4 times in drug-resistant tumor cells compared with drug-sensitive cells. GEM(Res) MCF7 cells had lower levels of the Notch-1-extracellular domain (NECD) and Notch trans-membrane intracellular domain (TMIC) than GEM(Sens) MCF7. The levels of Numb, and Numb-L-[P]-Ser(265) were similar in GEM(Res) and GEM(Sens) MCF7 cells. Only the levels of Numb-L (long)-Ser(295) decreased slightly. This finding suggests that Notch-1 cleavage to TMIC is inhibited in GEM(Res) MCF7 cells. PBMC activated by natural immunogenic peptides Notch-1 (2112-2120) and Numb-1 (87-95) eliminated NICD(positive), CD24(hi) CD24(lo) MCF7 cells. It is likely that the immunogenic Numb-1 peptide in MCF7 cells originated from Numb, [P]-lated by an unknown kinase, because staurosporine but not wortmannin and MAPK-inhibitors decreased peptide presentation. Numb and Notch are antagonistic proteins which degrade each other to stop and activate cell proliferation, respectively. Their peptides are presented alternatively. Targeting both antagonistic proteins should be useful to prevent metastases in patients whose tumors are resistant to conventional treatments.
authors.affiliation: Department of Gynecologic Oncology, The University of Texas M. D. Anderson Cancer Center, Houston, TX 77030, USA. mine@med.kurume-u.ac.jp
authors.forename: Takashi
authors.initials: T
authors.lastname: Mine
authors.forename: Satoko
authors.initials: S
authors.lastname: Matsueda
authors.forename: Yufeng
authors.initials: Y
authors.lastname: Li
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Tokumitsu
authors.forename: Hui
authors.initials: H
authors.lastname: Gao
authors.forename: Cristopher
authors.initials: C
authors.lastname: Danes
authors.forename: Kwong-Kwok
authors.initials: KK
authors.lastname: Wong
authors.forename: Xinhui
authors.initials: X
authors.lastname: Wang
authors.forename: Soldano
authors.initials: S
authors.lastname: Ferrone
authors.forename: Constantin G
authors.initials: CG
authors.lastname: Ioannides
mesh_terms: D016588:Anticarcinogenic Agents / Q000494:pharmacology; D014408:Biomarkers, Tumor / Q000276:immunology / Q000378:metabolism; D001943:Breast Neoplasms / Q000276:immunology* / Q000378:metabolism; D051927:CD24 Antigen / Q000276:immunology; D018414:CD8-Positive T-Lymphocytes / Q000187:drug effects / Q000276:immunology*; D045744:Cell Line, Tumor; D019008:Drug Resistance, Neoplasm / Q000187:drug effects / Q000502:physiology; D005260:Female; D058851:GPI-Linked Proteins; D006801:Humans; D018960:Hyaluronan Receptors / Q000276:immunology; D016233:Immunotherapy, Active; D016219:Immunotherapy, Adoptive; D036341:Intercellular Signaling Peptides and Proteins / Q000276:immunology / Q000378:metabolism; D007371:Interferon-gamma / Q000096:biosynthesis / Q000276:immunology; D007963:Leukocytes, Mononuclear / Q000187:drug effects / Q000276:immunology / Q000378:metabolism; D008213:Lymphocyte Activation; D008565:Membrane Proteins / Q000276:immunology* / Q000378:metabolism; D014411:Neoplastic Stem Cells / Q000276:immunology* / Q000378:metabolism; D009419:Nerve Tissue Proteins / Q000276:immunology* / Q000378:metabolism; D010051:Ovarian Neoplasms / Q000276:immunology / Q000378:metabolism; D010455:Peptides / Q000276:immunology; D051881:Receptor, Notch1 / Q000276:immunology / Q000378:metabolism*
chemical_list: D016588:Anticarcinogenic Agents; D014408:Biomarkers, Tumor; D051927:CD24 Antigen; D058851:GPI-Linked Proteins; D018960:Hyaluronan Receptors; D036341:Intercellular Signaling Peptides and Proteins; D008565:Membrane Proteins; C497761:NOTCH1 protein, human; D009419:Nerve Tissue Proteins; C113026:NUMB protein, human; D010455:Peptides; D051881:Receptor, Notch1; C423230:ULBP2 protein, human; D007371:Interferon-gamma