title: Gravity-sensitive signaling drives 3-dimensional formation of multicellular thyroid cancer spheroids.
abstract: This study focused on the effects induced by a random positioning machine (RPM) on FTC-133 thyroid cancer cells and evaluated signaling elements involved in 3-dimensional multicellular tumor spheroid (MCTS) formation. The cells were cultured on the RPM, a device developed to simulate microgravity, and under static 1-g conditions. After 24 h on the RPM, MCTSs swimming in culture supernatants were found, in addition to growth of adherent (AD) cells. Cells grown on the RPM showed higher levels of NF-κB p65 protein and apoptosis than 1-g controls, a result also found earlier in endothelial cells. Employing microarray analysis, we found 487 significantly regulated transcripts belonging not only to the apoptosis pathway but also to other biological processes. Selected transcripts were analyzed with quantitative real-time PCR using the same samples. Compared with 1-g IL-6, IL-8, CD44, and OPN were significantly up-regulated in AD cells but not in MCTSs, while ERK1/2, CAV2, TLN1, and CTGF were significantly down-regulated in AD cells. Simultaneously, the expression of ERK2, IL-6, CAV2, TLN1, and CTGF was reduced in MCTSs. IL-6 protein expression and secretion mirrored its gene expression. Thus, we concluded that the signaling elements IL-6, IL-8, OPN, TLN1, and CTGF are involved with NF-κB p65 in RPM-dependent thyroid carcinoma cell spheroid formation.
authors.affiliation: Department of Nuclear Medicine, University of Regensburg, Regensburg, Germany.
authors.forename: Jirka
authors.initials: J
authors.lastname: Grosse
authors.forename: Markus
authors.initials: M
authors.lastname: Wehland
authors.forename: Jessica
authors.initials: J
authors.lastname: Pietsch
authors.forename: Herbert
authors.initials: H
authors.lastname: Schulz
authors.forename: Katrin
authors.initials: K
authors.lastname: Saar
authors.forename: Norbert
authors.initials: N
authors.lastname: Hübner
authors.forename: Christoph
authors.initials: C
authors.lastname: Eilles
authors.forename: Johann
authors.initials: J
authors.lastname: Bauer
authors.forename: Khalil
authors.initials: K
authors.lastname: Abou-El-Ardat
authors.forename: Sarah
authors.initials: S
authors.lastname: Baatout
authors.forename: Xiao
authors.initials: X
authors.lastname: Ma
authors.forename: Manfred
authors.initials: M
authors.lastname: Infanger
authors.forename: Ruth
authors.initials: R
authors.lastname: Hemmersbach
authors.forename: Daniela
authors.initials: D
authors.lastname: Grimm
mesh_terms: D017209:Apoptosis / Q000235:genetics; D015153:Blotting, Western; D018929:Cell Culture Techniques / Q000379:methods; D045744:Cell Line, Tumor; D016000:Cluster Analysis; D055513:Connective Tissue Growth Factor / Q000235:genetics / Q000378:metabolism; D048049:Extracellular Signal-Regulated MAP Kinases / Q000235:genetics / Q000378:metabolism; D020869:Gene Expression Profiling; D015972:Gene Expression Regulation, Neoplastic; D006112:Gravitation; D006801:Humans; D018960:Hyaluronan Receptors / Q000235:genetics / Q000378:metabolism; D015850:Interleukin-6 / Q000235:genetics / Q000378:metabolism; D016209:Interleukin-8 / Q000235:genetics / Q000378:metabolism; D008297:Male; D020411:Oligonucleotide Array Sequence Analysis; D053495:Osteopontin / Q000235:genetics / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015398:Signal Transduction; D018874:Spheroids, Cellular / Q000378:metabolism* / Q000473:pathology; D016608:Talin / Q000235:genetics / Q000378:metabolism; D013964:Thyroid Neoplasms / Q000235:genetics* / Q000378:metabolism / Q000473:pathology; D051996:Transcription Factor RelA / Q000235:genetics / Q000378:metabolism
chemical_list: C526317:CCN2 protein, human; D018960:Hyaluronan Receptors; D015850:Interleukin-6; D016209:Interleukin-8; C518832:TLN1 protein, human; D016608:Talin; D051996:Transcription Factor RelA; D053495:Osteopontin; D055513:Connective Tissue Growth Factor; D048049:Extracellular Signal-Regulated MAP Kinases