title: Antisense inhibition of microRNA-21 and microRNA-221 in tumor-initiating stem-like cells modulates tumorigenesis, metastasis, and chemotherapy resistance in pancreatic cancer.
abstract: Our preliminary studies identified a small population side population (SP) cells in pancreatic cancer cells with stem cell-like properties, which were able to induce fast and aggressive tumor formation in nude mice. Gene expression analysis showed a significant difference in the expression of more than 1,300 genes in SP cells, among which a highly significant difference in microRNA expression of miR-21 and miR-221 between SP and NSP cells was identified. SP cells were identified and characterized by flow cytometry using Hoechst 33342 dye staining from a highly metastatic human pancreatic cancer cell line (L3.6pl). Antagomir transfection was performed using miRNA-21 and miRNA-221 antisense oligonucleotides (ASOs) and followed by detection of cell apoptosis, cell cycle progression, chemosensitivity, and invasion. Sorted SP cells from gemcitabine-resistant L3.6pl cells (L3.6pl(Gres)-SP) cells were orthotopically implanted in nude mice with or without miRNA-21 and miRNA-221 ASOs mono- and combination therapy. The administration of antagomir-21 and antagomir-221 significantly reduced the SP cell fraction, decreased SP cell differentiation, and downstream gene regulation, and thereby induced reduction of L3.6pl cell proliferation, invasion, and chemoresistance against gemcitabine and 5-Fluorouracil. Combination of ASOs therapy against miRNA-21 and miRNA-221 significantly inhibited primary tumor growth and metastasis compared to single antagomir treatment, especially, in L3.6plGres-SP-induced pancreatic tumor growth in vivo. These findings further indicate that the inhibition of miR-21 and miR-221 appear particularly suitable to target stem-like subpopulations and address their specific biological function to promote tumor progression in pancreatic cancer.
authors: Department of General, Visceral und Vascular Surgery, Otto-von-Guericke University, Leipziger Strasse 44, 39120, Magdeburg, Germany. zyberry@gmail.com.
authors: Yue
authors: Y
authors: Zhao
authors: Department of General, Visceral und Vascular Surgery, University of Munich, Campus Grosshadern, Munich, Germany.
authors: Lu
authors: L
authors: Zhao
authors: Department of General, Visceral und Vascular Surgery, University of Munich, Campus Grosshadern, Munich, Germany.
authors: Ivan
authors: I
authors: Ischenko
authors: Department of General, Visceral und Vascular Surgery, University of Munich, Campus Grosshadern, Munich, Germany.
authors: Qi
authors: Q
authors: Bao
authors: Department of General, Visceral und Vascular Surgery, University of Munich, Campus Grosshadern, Munich, Germany.
authors: Bettina
authors: B
authors: Schwarz
authors: Department of General, Visceral und Vascular Surgery, University of Munich, Campus Grosshadern, Munich, Germany.
authors: Hanno
authors: H
authors: Nieß
authors: Department of General, Visceral und Vascular Surgery, Otto-von-Guericke University, Leipziger Strasse 44, 39120, Magdeburg, Germany.
authors: Yan
authors: Y
authors: Wang
authors: Department of General, Visceral und Vascular Surgery, University of Munich, Campus Grosshadern, Munich, Germany.
authors: Andrea
authors: A
authors: Renner
authors: Institute of Molecular Immunology, Helmholtz Center Munich, Munich, Germany.
authors: Josef
authors: J
authors: Mysliwietz
authors: Department of General, Visceral und Vascular Surgery, University of Munich, Campus Grosshadern, Munich, Germany.
authors: Karl-Walter
authors: KW
authors: Jauch
authors: Clinical Biochemistry Group, Medizinische Klinik und Poliklinik IV, University of Munich, Munich, Germany.
authors: Peter J
authors: PJ
authors: Nelson
authors: Institute of Molecular Immunology, Helmholtz Center Munich, Munich, Germany.
authors: Joachim W
authors: JW
authors: Ellwart
authors: Department of General, Visceral und Vascular Surgery, Otto-von-Guericke University, Leipziger Strasse 44, 39120, Magdeburg, Germany.
authors: Christiane J
authors: CJ
authors: Bruns
authors: Department of General, Visceral und Vascular Surgery, Otto-von-Guericke University, Leipziger Strasse 44, 39120, Magdeburg, Germany. peter.camaj@med.ovgu.de.
authors: Peter
authors: P
authors: Camaj
mesh_terms: D000818:Animals; D063646:Carcinogenesis / Q000235:genetics; D002454:Cell Differentiation / Q000235:genetics; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000235:genetics; D019008:Drug Resistance, Neoplasm; D006801:Humans; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D008819:Mice, Nude; D035683:MicroRNAs / Q000037:antagonists & inhibitors* / Q000235:genetics / Q000378:metabolism; D009362:Neoplasm Metastasis; D014411:Neoplastic Stem Cells / Q000473:pathology / Q000502:physiology*; D016376:Oligonucleotides, Antisense / Q000008:administration & dosage* / Q000235:genetics; D010190:Pancreatic Neoplasms / Q000235:genetics* / Q000473:pathology / Q000628:therapy*; D014162:Transfection; D015854:Up-Regulation; D023041:Xenograft Model Antitumor Assays
chemical_list: D035683:MicroRNAs; D016376:Oligonucleotides, Antisense; C524940:MIRN21 microRNA, human; C525709:MIR221, human
keywords: Chemotherapy resistance; Metastasis; Pancreatic cancer; Side population; Tumorigenesis; miRNAs
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