title: MicroRNAs as potential biomarkers for VERO cell tumorigenicity.
abstract: MicroRNA expression appears to capture the process of neoplastic development in vitro in the VERO line of African green monkey kidney (AGMK) cells (Teferedegne et al. PLoS One 2010;5(12):e14416). In that study, specific miRNA signatures were correlated with the transition, during serial tissue-culture passage, of low-density passaged 10-87 VERO cells from a non-tumorigenic phenotype at passage (p) 148 to a tumorigenic phenotype at p256. In the present study, six miRNAs (miR-376a, miR-654-3p, miR-543, miR-299-3p, miR-134 and miR-369-3p) were chosen from the identified signature miRNAs for evaluation of their use as potential biomarkers to track the progression of neoplastic development in VERO cells. Cells from the 10-87 VERO cell line at passage levels from p148 to p256 were inoculated into newborn and adult athymic nude mice. No tumors were observed in animals inoculated with cells from p148 to p186. In contrast, tumor incidences of 20% developed only in newborn mice that received 10-87 VERO cells at p194, p234 and p256. By qPCR profiling of the signature miRNAs of 10-87 VERO cells from these cell banks, we identified p194 as the level at which signature miRNAs elevated concurrently with the acquisition of tumorigenic phenotype with similar levels expressed beyond this passage. In wound-healing assays at 10-passage intervals between p150 to p250, the cells displayed a progressive increase in migration from p165 to p186; beginning at p194 and higher passages thereafter, the cells exhibited the highest rates of migration. By qPCR analysis, the same signature miRNAs were overexpressed with concomitant acquisition of the tumorigenic phenotype in another lineage of 10-87 VERO cells passaged independently at high density. Correlation between the passages at which the cells expressed a tumorigenic phenotype and the passages representing peaks in expression levels of signature miRNAs indicates that these miRNAs are potential biomarkers for the expression of the VERO cell tumorigenic phenotype.
authors.affiliation: Laboratory of DNA Viruses, Division of Viral Products, Office of Vaccines Research and Review, Center for Biologics Evaluation and Research, Food and Drug Administration, Bethesda, MD 20892, United States. Electronic address: belete.teferedegne@fda.hhs.gov.
authors.forename: Belete
authors.initials: B
authors.lastname: Teferedegne
authors.affiliation: Laboratory of DNA Viruses, Division of Viral Products, Office of Vaccines Research and Review, Center for Biologics Evaluation and Research, Food and Drug Administration, Bethesda, MD 20892, United States.
authors.forename: Juliete
authors.initials: J
authors.lastname: Macauley
authors.affiliation: Laboratory of DNA Viruses, Division of Viral Products, Office of Vaccines Research and Review, Center for Biologics Evaluation and Research, Food and Drug Administration, Bethesda, MD 20892, United States.
authors.forename: Gideon
authors.initials: G
authors.lastname: Foseh
authors.affiliation: Laboratory of Method Development, Division of Viral Products, Office of Vaccines Research and Review, Center for Biologics Evaluation and Research, Food and Drug Administration, Bethesda, MD 20892, United States.
authors.forename: Eugenia
authors.initials: E
authors.lastname: Dragunsky
authors.affiliation: Laboratory of Method Development, Division of Viral Products, Office of Vaccines Research and Review, Center for Biologics Evaluation and Research, Food and Drug Administration, Bethesda, MD 20892, United States.
authors.forename: Konstantin
authors.initials: K
authors.lastname: Chumakov
authors.affiliation: Laboratory of DNA Viruses, Division of Viral Products, Office of Vaccines Research and Review, Center for Biologics Evaluation and Research, Food and Drug Administration, Bethesda, MD 20892, United States.
authors.forename: Haruhiko
authors.initials: H
authors.lastname: Murata
authors.affiliation: Laboratory of DNA Viruses, Division of Viral Products, Office of Vaccines Research and Review, Center for Biologics Evaluation and Research, Food and Drug Administration, Bethesda, MD 20892, United States.
authors.forename: Keith
authors.initials: K
authors.lastname: Peden
authors.affiliation: Laboratory of DNA Viruses, Division of Viral Products, Office of Vaccines Research and Review, Center for Biologics Evaluation and Research, Food and Drug Administration, Bethesda, MD 20892, United States. Electronic address: andrew.lewis@fda.hhs.gov.
authors.forename: Andrew M
authors.initials: AM
authors.lastname: Lewis
mesh_terms: D000818:Animals; D015415:Biomarkers; D002465:Cell Movement; D002471:Cell Transformation, Neoplastic / Q000235:genetics*; D002522:Chlorocebus aethiops; D008819:Mice, Nude; D035683:MicroRNAs / Q000235:genetics*; D010641:Phenotype; D014709:Vero Cells / Q000166:cytology*
chemical_list: D015415:Biomarkers; D035683:MicroRNAs
keywords: African green monkey kidney (VERO) cells; Biomarkers; Neoplastic development; Transformation; Tumorigenicity; miRNA