Title:MARK1 is a Novel Target for miR-125a-5p: Implications for Cell Migration in Cervical Tumor Cells
VOLUME: 7 ISSUE: 1
Author(s):Martínez-Acuna Natalia, Gonzalez-Torres Alejandro, Tapia-Vieyra Juana Virginia and Luis Marat Alvarez-Salas*
Affiliation:Laboratorio de Terapia Genica, Departamento de Genetica y Biologia Molecular, Centro de Investigacion y de Estudios Avanzados del I.P.N., Ciudad de Mexico, Laboratorio de Terapia Genica, Departamento de Genetica y Biologia Molecular, Centro de Investigacion y de Estudios Avanzados del I.P.N., Ciudad de Mexico, Laboratorio de Terapia Genica, Departamento de Genetica y Biologia Molecular, Centro de Investigacion y de Estudios Avanzados del I.P.N., Ciudad de Mexico, Laboratorio de Terapia Genica, Departamento de Genetica y Biologia Molecular, Centro de Investigacion y de Estudios Avanzados del I.P.N., Ciudad de Mexico
Keywords:Cell migration, cervical cancer, MARK1, microRNAs, miR-125a-5p, tumor.
Abstract:Background: Aberrant miRNA expression is associated with the development of several
diseases including cervical cancer. Dysregulation of miR-125a-5p is present in a plethora of tumors,
but its role in cervical cancer is not well understood.
Objective: The aim was to analyze the expression profile of miR-125a-5p in tumor and immortal cell
lines with further target prediction, validation and function analysis.
Methods: MiR-125a-5p expression was determined by real-time RT-PCR from nine cervical cell
lines. In silico tools were used to find target transcripts with an miR-125-5p complementary site
within the 3'UTR region. Further target selection was based on gene ontology annotation and ΔG
analysis. Target validation was performed by transfection of synthetic miR-125a-5p mimics and luciferase
assays. Functional evaluation of miR-125a-5p on migration was performed by transwell migration
assays.
Results: Differential miR-125a-5p expression was observed between immortal and tumor cells regardless
of the human papillomavirus (HPV) content. Thermodynamic and ontological analyses showed
Microtubule-Affinity-Regulating Kinase1 (MARK1) as a putative target for miR-125a-5p. An inverse
correlation was observed among miR-125a-5p expression and MARK1 protein levels in tumor but not
in immortal cells. Luciferase assays showed direct miR-125a-5p regulation over MARK1 through recognition
of a predicted target site within the 3'-UTR. HeLa and C-33A cervical tumor cells enhanced
migration after transfection with miR-125a-5p mimics and stimulation of cell migration was reproduced
by siRNA-mediated inhibition of MARK1.
Conclusion: The results showed MARK1 as a novel functional target for miR-125a-5p with implications
on cell migration of tumor cervical cancer cells.
