title: Multiparameter analyses of three-dimensionally cultured tumor spheroids based on respiratory activity and comprehensive gene expression profiles.
abstract: Multicellular spheroids of human breast cancer cells (MCF-7) formed with two different three-dimensional (3D) culture methods were evaluated in detail on the basis of respiratory activity and high-throughput gene expression analysis. The spheroids formed with poly(dimethylsiloxane) (PDMS) microwell arrays indicated significant restriction of the spheroid size, whereas their respiratory activity was 2-fold greater than that formed with the hanging drop culture method. Fluidigm BioMark dynamic array was used for comprehensive and quantitative real-time polymerase chain reaction (qRT-PCR) analysis on the samples whose respiratory activity had been measured. Genes involved in cellular senescence and glucose metabolism indicated significantly higher values for the PDMS microwell culture method than for the hanging drop culture method (P<0.05). Interestingly, samples formed with the PDMS microwell culture method showed stronger responses for glycolysis than those formed with the hanging drop method. These results illustrate the power of multiparameter analysis to characterize multicellular spheroids cultured in different microenvironments even if they have the same morphology.
authors.affiliation: Graduate School of Environmental Studies, Tohoku University, Sendai 980-8579, Japan.
authors.forename: Yuanshu
authors.initials: Y
authors.lastname: Zhou
authors.forename: Toshiharu
authors.initials: T
authors.lastname: Arai
authors.forename: Yoshiko
authors.initials: Y
authors.lastname: Horiguchi
authors.forename: Kosuke
authors.initials: K
authors.lastname: Ino
authors.forename: Tomokazu
authors.initials: T
authors.lastname: Matsue
authors.forename: Hitoshi
authors.initials: H
authors.lastname: Shiku
mesh_terms: D001943:Breast Neoplasms / Q000378:metabolism; D018929:Cell Culture Techniques / Q000379:methods*; D045744:Cell Line, Tumor; D004734:Energy Metabolism; D005260:Female; D015972:Gene Expression Regulation, Neoplastic / Q000502:physiology; D006801:Humans; D010101:Oxygen Consumption / Q000502:physiology; D040081:Protein Array Analysis / Q000379:methods; D059467:Transcriptome / Q000502:physiology*
keywords: High-throughput qPCR; Multicellular tumor spheroids; PDMS microwell array; Respiratory activity