title: miR-24 Inhibition Increases Menin Expression and Decreases Cholangiocarcinoma Proliferation.
abstract: Menin (MEN1) is a tumor-suppressor protein in neuroendocrine tissue. Therefore, we tested the novel hypothesis that menin regulates cholangiocarcinoma proliferation. Menin and miR-24 expression levels were measured in the following intrahepatic and extrahepatic cholangiocarcinoma (CCA) cell lines, Mz-ChA-1, TFK-1, SG231, CCLP, HuCCT-1, and HuH-28, as well as the nonmalignant human intrahepatic biliary line, H69. miR-24 miRNA and menin protein levels were manipulated in vitro in Mz-ChA-1 cell lines. Markers of proliferation and angiogenesis (Ki-67, vascular endothelial growth factors A/C, vascular endothelial growth factor receptors 2/3, angiopoietin 1/2, and angiopoietin receptors 1/2) were evaluated. Mz-ChA-1 cells were injected into the flanks of nude mice and treated with miR-24 inhibitor or inhibitor scramble. Menin expression was decreased in advanced CCA specimens, whereas miR-24 expression was increased in CCA. Menin overexpression decreased proliferation, angiogenesis, migration, and invasion. Inhibition of miR-24 increased menin protein expression while decreasing proliferation, angiogenesis, migration, and invasion. miR-24 was shown to negatively regulate menin expression by luciferase assay. Tumor burden and expression of proliferative and angiogenic markers was decreased in the miR-24 inhibited tumor group compared to controls. Interestingly, treated tumors were more fibrotic than the control group. miR-24-dependent expression of menin may be important in the regulation of nonmalignant and CCA proliferation and may be an additional therapeutic tool for managing CCA progression.
authors.affiliation: Department of Medicine, Baylor Scott & White and Texas A&M University Health Science Center, Temple, Texas; Division of Gastroenterology and Medical Physiology, Baylor Scott & White and Texas A&M University Health Science Center, Temple, Texas.
authors.forename: Laurent
authors.initials: L
authors.lastname: Ehrlich
authors.affiliation: Division of Surgery, Baylor Scott & White and Texas A&M University Health Science Center, Temple, Texas.
authors.forename: Chad
authors.initials: C
authors.lastname: Hall
authors.affiliation: Department of Medicine, Baylor Scott & White and Texas A&M University Health Science Center, Temple, Texas.
authors.forename: Julie
authors.initials: J
authors.lastname: Venter
authors.affiliation: Division of Gastroenterology and Medical Physiology, Baylor Scott & White and Texas A&M University Health Science Center, Temple, Texas.
authors.forename: David
authors.initials: D
authors.lastname: Dostal
authors.affiliation: Department of Medicine and Surgery, Program for Autoimmune Liver Diseases, International Center for Digestive Diseases, University of Milan-Bicocca, Milan, Italy; Center for Autoimmune Liver Diseases, Humanitas Clinical and Research Center, Rozzano (Milan), Italy.
authors.forename: Francesca
authors.initials: F
authors.lastname: Bernuzzi
authors.affiliation: Department of Medicine and Surgery, Program for Autoimmune Liver Diseases, International Center for Digestive Diseases, University of Milan-Bicocca, Milan, Italy; Center for Autoimmune Liver Diseases, Humanitas Clinical and Research Center, Rozzano (Milan), Italy.
authors.forename: Pietro
authors.initials: P
authors.lastname: Invernizzi
authors.affiliation: Department of Medicine, Baylor Scott & White and Texas A&M University Health Science Center, Temple, Texas; Division of Gastroenterology and Medical Physiology, Baylor Scott & White and Texas A&M University Health Science Center, Temple, Texas; Research Section, Central Texas Veterans Health Care System, Baylor Scott & White and Texas A&M University Health Science Center, Temple, Texas; Baylor Scott & White Digestive Disease Research Center, Baylor Scott & White and Texas A&M University Health Science Center, Temple, Texas.
authors.forename: Fanyin
authors.initials: F
authors.lastname: Meng
authors.affiliation: Division of Gastroenterology and Medical Physiology, Baylor Scott & White and Texas A&M University Health Science Center, Temple, Texas.
authors.forename: Jerome P
authors.initials: JP
authors.lastname: Trzeciakowski
authors.affiliation: Department of Medicine, Baylor Scott & White and Texas A&M University Health Science Center, Temple, Texas.
authors.forename: Tianhao
authors.initials: T
authors.lastname: Zhou
authors.affiliation: Research Section, Central Texas Veterans Health Care System, Baylor Scott & White and Texas A&M University Health Science Center, Temple, Texas.
authors.forename: Holly
authors.initials: H
authors.lastname: Standeford
authors.affiliation: Department of Medicine, Baylor Scott & White and Texas A&M University Health Science Center, Temple, Texas; Research Section, Central Texas Veterans Health Care System, Baylor Scott & White and Texas A&M University Health Science Center, Temple, Texas; Baylor Scott & White Digestive Disease Research Center, Baylor Scott & White and Texas A&M University Health Science Center, Temple, Texas.
authors.forename: Gianfranco
authors.initials: G
authors.lastname: Alpini
authors.affiliation: Division of Surgery, Baylor Scott & White and Texas A&M University Health Science Center, Temple, Texas.
authors.forename: Terry C
authors.initials: TC
authors.lastname: Lairmore
authors.affiliation: Department of Medicine, Baylor Scott & White and Texas A&M University Health Science Center, Temple, Texas; Research Section, Central Texas Veterans Health Care System, Baylor Scott & White and Texas A&M University Health Science Center, Temple, Texas; Baylor Scott & White Digestive Disease Research Center, Baylor Scott & White and Texas A&M University Health Science Center, Temple, Texas. Electronic address: sglaser@medicine.tamhsc.edu.
authors.forename: Shannon
authors.initials: S
authors.lastname: Glaser
mesh_terms: D000368:Aged; D043925:Angiogenesis Inducing Agents / Q000378:metabolism; D000818:Animals; D001650:Bile Duct Neoplasms / Q000473:pathology; D001652:Bile Ducts / Q000473:pathology; D001706:Biopsy; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000235:genetics; D018281:Cholangiocarcinoma / Q000235:genetics* / Q000473:pathology*; D015536:Down-Regulation / Q000235:genetics; D015972:Gene Expression Regulation, Neoplastic; D006801:Humans; D008297:Male; D008807:Mice, Inbred BALB C; D008819:Mice, Nude; D035683:MicroRNAs / Q000037:antagonists & inhibitors* / Q000378:metabolism; D011518:Proto-Oncogene Proteins / Q000235:genetics* / Q000378:metabolism; D023041:Xenograft Model Antitumor Assays
chemical_list: D043925:Angiogenesis Inducing Agents; C105373:MEN1 protein, human; C543845:MIRN24 microRNA, human; D035683:MicroRNAs; D011518:Proto-Oncogene Proteins