OBJECTIVE: A new interleukin-6 (IL-6)-dependent plasma cell leukemia cell line UHKT-944 was established from bone marrow cells derived from a 55-yr-old man with plasma cell leukemia. RESULTS: The cell line possesses phenotypic characteristics of plasma cells including the production of a monoclonal immunoglobulin IgA1-kappa. VH3-9 region of IgVH genes was rearranged and somatically hypermutated. The UHKT-944 cells were found to be negative for most of tested B-cell, T-cell, and myeloid markers. According to cytogenetic analysis, the cells were classified as near tetraploid with several numerical and structural abnormalities including the t(14;20) involving IgH locus. CONCLUSION: The established permanent plasma cell leukemia cell line is a suitable model for the study of cellular and molecular mechanisms of pathogenesis of this rare malignant disease.

• Klíčová slova
• human plasma cell line, interleukin-6 (IL-6), multiple myeloma, plasma cell, plasma cell leukemia,
• MeSH
• biologické markery MeSH
• cytogenetické vyšetření MeSH
• imunofenotypizace MeSH
• imunoglobuliny genetika   metabolismus   MeSH
• lidé středního věku MeSH
• lidé MeSH
• nádorové buněčné linie MeSH
• plazmocelulární leukemie diagnóza   metabolismus   patologie   MeSH
• proliferace buněk MeSH
• Check Tag
• lidé středního věku MeSH
• lidé MeSH
• mužské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• kazuistiky MeSH
• práce podpořená grantem MeSH
• Názvy látek
• biologické markery MeSH
• imunoglobuliny MeSH

UNLABELLED: We established and characterized a new IL-6 dependent multiple myeloma (MM) cell line UHKT-893 from the bone marrow of a relapsed 57-year-old woman. RESULTS: Using nephelometry, cells with plasma cell phenotype and morphology were found to secrete IgG and free kappa (κ)-light chain of immunoglobulin. κ-Light chain was also recognized intracellularly by flow cytometry and by mass spectrometry. VH4-39 region of IgVH genes was rearranged and somatically hypermutated. Cytogenetic analysis of cells revealed new chromosome abnormalities in all breakpoints unique in both MM patients and cell lines - t(1;6), t(1;11), t(5;15), t(5;21), +der(11;15) and der(16). IL-6 independent subline UHKT-893a was established by adaptation to descending IL-6 concentration, while the original cell line keeps on maintaining its IL-6 dependency. CONCLUSION: The cell line provides a suitable material for cellular and molecular studies of tumor abnormalities, with potentially unique mutagenic features of myeloma disease. It may be utilized for human hybridoma construction and vaccine development. Both IL-6 dependent and independent cell clones represent an important model for studies of myeloma cell growth and resistance emerging during targeted therapy.

• MeSH
• buňky kostní dřeně účinky léků   patologie   fyziologie   MeSH
• cytogenetické vyšetření MeSH
• interleukin-6 farmakologie   MeSH
• lidé středního věku MeSH
• lidé MeSH
• mnohočetný myelom patologie   MeSH
• nádorové buněčné linie MeSH
• primární buněčná kultura * metody   MeSH
• proliferace buněk účinky léků   MeSH
• recidiva MeSH
• Check Tag
• lidé středního věku MeSH
• lidé MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• interleukin-6 MeSH

BACKGROUND/AIMS: Alteration of cancer cell redox status has been recognized as a promising therapeutic implication. In recent years, the emerged field of non-thermal plasma (NTP) has shown considerable promise in various biomedical applications, including cancer therapy. However, understanding the molecular mechanisms procuring cellular responses remains incomplete. Thus, the aim of this study was a rigorous biochemical analysis of interactions between NTP and liver cancer cells. METHODS: The concept was validated using three different cell lines. We provide several distinct lines of evidence to support our findings; we use various methods (epifluorescent and confocal microscopy, clonogenic and cytotoxicity assays, Western blotting, pharmacological inhibition studies, etc.). RESULTS: We assessed the influence of NTP on three human liver cancer cell lines (Huh7, Alexander and HepG2). NTP treatment resulted in higher anti-proliferative effect against Alexander and Huh7 relative to HepG2. Our data clearly showed that the NTP-mediated alternation of mitochondrial membrane potential and dynamics led to ROS-mediated apoptosis in Huh7 and Alexander cells. Interestingly, plasma treatment resulted in p53 down-regulation in Huh7 cells. High levels of Bcl-2 protein expression in HepG2 resulted in their resistance in response to oxidative stress- mediated by plasma. CONCLUSION: We show thoroughly time- and dose-dependent kinetics of ROS accumulation in HCC cells. Furthermore, we show nuclear compartmentalization of the superoxide anion triggered by NTP. NTP induced apoptotic death in Huh7 liver cancer cells via simultaneous downregulation of mutated p53, pSTAT1 and STAT1. Contrary, hydrogen peroxide treatment results in autophagic cell death. We disclosed detailed mechanisms of NTP-mediated alteration of redox signalling in liver cancer cells.

• Klíčová slova
• Apoptosis, Non-thermal plasma, Oxidative stress, Reactive oxygen species, Stress resistance, p53,
• MeSH
• buněčná smrt účinky léků   MeSH
• buňky Hep G2 MeSH
• down regulace účinky léků   MeSH
• hepatocelulární karcinom farmakoterapie   genetika   metabolismus   patologie   MeSH
• lidé MeSH
• nádorový supresorový protein p53 biosyntéza   genetika   MeSH
• nádory jater farmakoterapie   genetika   metabolismus   patologie   MeSH
• oxidace-redukce účinky léků   MeSH
• plazmové plyny farmakologie   MeSH
• reaktivní formy kyslíku metabolismus   MeSH
• regulace genové exprese u nádorů účinky léků   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• nádorový supresorový protein p53 MeSH
• plazmové plyny MeSH
• reaktivní formy kyslíku MeSH
• TP53 protein, human MeSH Prohlížeč

Circulating cell-free DNA (cfDNA) may be involved in immune response regulation. We studied the variations in abundance of telomeric sequences in plasma and serum in young healthy volunteers and the ability of cfDNA contained in these samples to co-activate the TNF-α m RNA expression in monocytes. We performed qPCR to determine relative telomere length (T/S ratios) in plasma, serum and whole blood of 36 volunteers. Using paired samples of plasma and serum and DNase treatment, we analysed the contribution of cfDNA to the co-activation of TNF-α mRNA expression in THP1 monocytic cell line. We found significant differences between paired plasma and serum samples in relative T/S ratios (median 1.38 ± 1.1 vs. 0.86 ± 0.25, respectively) and in total amounts of cfDNA and in estimated total amounts of telomeres which were significantly higher in serum than in plasma. TNF-α mRNA expression in THP1 cells increased significantly after DNase treatment of all samples used for stimulation. The highest TNF-α mRNA expressions were observed after stimulation with DNase treated serum samples. Our results suggest that the different content of telomeric sequences in plasma and serum may contribute to the tuning of immune response. Further studies of this interesting phenomenon are needed.

• MeSH
• deoxyribonukleasy metabolismus   MeSH
• homeostáza telomer MeSH
• imunita MeSH
• imunomodulace MeSH
• krevní plazma imunologie   metabolismus   MeSH
• lidé MeSH
• mladý dospělý MeSH
• monocyty fyziologie   MeSH
• sérum imunologie   metabolismus   MeSH
• telomery genetika   MeSH
• THP-1 buňky MeSH
• TNF-alfa genetika   metabolismus   MeSH
• upregulace MeSH
• volné cirkulující nukleové kyseliny genetika   imunologie   metabolismus   MeSH
• zdraví dobrovolníci pro lékařské studie MeSH
• Check Tag
• lidé MeSH
• mladý dospělý MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• srovnávací studie MeSH
• Názvy látek
• deoxyribonukleasy MeSH
• TNF-alfa MeSH
• volné cirkulující nukleové kyseliny MeSH

Our previously-obtained impressive results of highly increased C2C12 mouse myoblast adhesion to amine plasma polymers (PPs) motivated current detailed studies of cell resistance to trypsinization, cell proliferation, motility, and the rate of attachment carried out for fibroblasts (LF), keratinocytes (HaCaT), rat vascular smooth muscle cells (VSMC), and endothelial cells (HUVEC, HSVEC, and CPAE) on three different amine PPs. We demonstrated the striking difference in the resistance to trypsin treatment between endothelial and non-endothelial cells. The increased resistance observed for the non-endothelial cell types was accompanied by an increased rate of cellular attachment, even though spontaneous migration was comparable to the control, i.e., to the standard cultivation surface. As demonstrated on LF fibroblasts, the resistance to trypsin was similar in serum-supplemented and serum-free media, i.e., medium without cell adhesion-mediating proteins. The increased cell adhesion was also confirmed for LF cells by an independent technique, single-cell force spectroscopy. This method, as well as the cell attachment rate, proved the difference among the plasma polymers with different amounts of amine groups, but other investigated techniques could not reveal the differences in the cell behaviour on different amine PPs. Based on all the results, the increased resistance to trypsinization of C2C12, LF, HaCaT, and VSMC cells on amine PPs can be explained most probably by a non-specific cell adhesion such as electrostatic interaction between the cells and amine groups on the material surface, rather than by the receptor-mediated adhesion through serum-derived proteins adsorbed on the PPs.

• MeSH
• aminy chemie   MeSH
• buněčná adheze účinky léků   MeSH
• buněčné linie MeSH
• lidé MeSH
• plazmové plyny chemie   MeSH
• polymery chemie   farmakologie   MeSH
• povrchové vlastnosti MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• aminy MeSH
• plazmové plyny MeSH
• polymery MeSH

Time-lapse cinemicrography was used to study the locomotory activity of sperical cells of a heteronuclear cell line VUP-1 originating from a malignant melanoma of chorioidea. Migration of cells was followed during the entire cell cycle and locomotion was analyzed in relation to the population density and to the morphological and functional characteristics of the leading undulating membrane. The variations in cellular motility were related to the morphology of the membranes and their possible functional regulation.

• MeSH
• buněčná membrána MeSH
• buněčné linie * MeSH
• epitelové buňky MeSH
• lidé MeSH
• melanom MeSH
• nádory choroidey MeSH
• pohyb buněk * MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH

Quaternary benzo[c]phenanthridine alkaloids (QBA), fagaronine (FA), sanguinarine (SA), chelerythrine (CHE) and the QBA extract from Macleya cordata (EX) exerted differential inhibitory effect on the hydrolytic activity of particular dipeptidyl peptidase (DPP)-like enzyme isolated from human blood plasma and from human and rat glioma cell lines. The low-MW form of DPP-IV-like enzyme activity, corresponding most probably to DPP-8, observed only in glioma cells but not in human plasma, was inhibited preferentially by SA, CHE and EX, and only slightly by FA. The alkaloid inhibitory effect was concentration-dependent in the range 25-150 mM and directly pH-related. In addition, a subtle but consistent inhibition of the intermediate-MW form of DPP-IV-like enzyme activity, ascribed to DPP-IV/CD26, observed only in human plasma and of the attractin (high-MW form of DPP-IV-like enzyme activity, expressed in U87 glioma cells) by the studied alkaloids was observed. We conclude that some of the QBA biological effects could be determined by tissue and cell type specific dipeptidyl peptidase IV-like molecules expression pattern.

• MeSH
• alkaloidy chemická syntéza   izolace a purifikace   farmakologie   MeSH
• benzofenantridiny MeSH
• dipeptidylpeptidasa 4 krev   izolace a purifikace   metabolismus   MeSH
• fenantridiny chemie   izolace a purifikace   farmakologie   MeSH
• gelová chromatografie metody   MeSH
• gliom enzymologie   patologie   MeSH
• inhibitory enzymů farmakologie   MeSH
• isochinoliny MeSH
• lidé MeSH
• nádorové buněčné linie enzymologie   MeSH
• Papaveraceae chemie   MeSH
• spektrofotometrie metody   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• alkaloidy MeSH
• benzofenantridiny MeSH
• chelerythrine MeSH Prohlížeč
• dipeptidylpeptidasa 4 MeSH
• fagaronine MeSH Prohlížeč
• fenantridiny MeSH
• inhibitory enzymů MeSH
• isochinoliny MeSH
• sanguinarine MeSH Prohlížeč

1. Breast cancer resistance protein (BCRP, ABCG2) is a drug efflux transporter that is believed to affect the drug disposition of several drugs and xenobiotics. In the present study, we evaluated the localization and functional expression of BCRP in the human choriocarcinoma cell line BeWo, an in vitro model of the human trophoblast, and compared it with the expression of P-glycoprotein (MDR1, ABCB1) as the most widely studied placental transporter. In addition, the expression of BCRP at the mRNA level was compared with that of MDR1 in the human term placenta. 2. Western blotting analysis revealed high endogenous expression of BCRP protein in BeWo cells. Using indirect immunofluorescence microscopy, we found that the BCRP transporter appears to be localized predominantly at the apical plasma membrane. Functional studies showed a significant effect of the BCRP inhibitors GF120918 (5 micromol/L) and Ko143 (1 micromol/L) on mitoxantrone accumulation and, thus, confirmed efflux activity of BCRP in BeWo cells. 3. Using absolute mRNA quantification with real-time reverse transcription-polymerase chain reaction, we found high expression of BCRP in BeWo cells, whereas no transcript of MDR1 (P-glycoprotein), the most extensively studied drug transporter, was detected. 4. In the human placenta, BCRP was localized predominantly in the syncytiotrophoblast layer; however, immunopositivity for the BXP-21 antibody was also observed in fetal vessels of the chorionic villi. The number of BCRP transcripts in the human term placenta was found to be more than 10-fold higher compared with the expression of MDR1. 5. In conclusion, we suggest that BeWo cells could serve as a suitable in vitro model to study trans-trophoblast transport of BCRP substrates and that placental BCRP can play an important role in preventing the accumulation of potentially toxic xenobiotics in the trophoblast cells.

• MeSH
• ABC transportér z rodiny G, člen 2 MeSH
• ABC transportéry antagonisté a inhibitory   genetika   metabolismus   MeSH
• akridiny farmakologie   MeSH
• biologický transport účinky léků   MeSH
• buněčná membrána metabolismus   MeSH
• buněčné linie MeSH
• choriokarcinom genetika   metabolismus   patologie   MeSH
• fluorescenční mikroskopie MeSH
• imunohistochemie MeSH
• indoly chemie   farmakologie   MeSH
• lidé MeSH
• messenger RNA genetika   metabolismus   MeSH
• mitoxantron farmakokinetika   MeSH
• nádorové buněčné linie MeSH
• nádorové proteiny antagonisté a inhibitory   genetika   metabolismus   MeSH
• P-glykoprotein genetika   metabolismus   MeSH
• placenta metabolismus   MeSH
• polymerázová řetězová reakce s reverzní transkripcí MeSH
• regulace genové exprese u nádorů * MeSH
• těhotenství MeSH
• tetrahydroisochinoliny farmakologie   MeSH
• western blotting MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• těhotenství MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• ABC transportér z rodiny G, člen 2 MeSH
• ABC transportéry MeSH
• ABCG2 protein, human MeSH Prohlížeč
• akridiny MeSH
• Elacridar MeSH Prohlížeč
• indoly MeSH
• messenger RNA MeSH
• mitoxantron MeSH
• nádorové proteiny MeSH
• P-glykoprotein MeSH
• tetrahydroisochinoliny MeSH
• tryptoquivaline MeSH Prohlížeč

The growing application of materials containing TiO2 particles has led to an increased risk of human exposure, while a gap in knowledge about the possible adverse effects of TiO2 still exists. In this work, TiO2 particles of rutile, anatase, and their commercial mixture were exposed to various environments, including simulated gastric fluids and human blood plasma (both representing in vivo conditions), and media used in in vitro experiments. Simulated body fluids of different compositions, ionic strengths, and pH were used, and the impact of the absence or presence of chosen enzymes was investigated. The physicochemical properties and agglomeration of TiO2 in these media were determined. The time dependent agglomeration of TiO2 related to the type of TiO2, and mainly to the type and composition of the environment that was observed. The presence of enzymes either prevented or promoted TiO2 agglomeration. TiO2 was also observed to exhibit concentration-dependent cytotoxicity. This knowledge about TiO2 behavior in all the abovementioned environments is critical when TiO2 safety is considered, especially with respect to the significant impact of the presence of proteins and size-related cytotoxicity.

• Klíčová slova
• TiO2 particles, agglomeration, plasma, proteins, simulated gastric fluids,
• MeSH
• buněčné linie MeSH
• dárci krve MeSH
• fibroblasty účinky léků   metabolismus   MeSH
• koncentrace vodíkových iontů MeSH
• kovové nanočástice škodlivé účinky   chemie   MeSH
• krevní plazma metabolismus   MeSH
• krystalizace MeSH
• kultivační média metabolismus   MeSH
• lidé MeSH
• myši MeSH
• osmolární koncentrace MeSH
• povrchové vlastnosti MeSH
• sliny metabolismus   MeSH
• titan škodlivé účinky   chemie   metabolismus   MeSH
• velikost částic MeSH
• viabilita buněk účinky léků   MeSH
• voda metabolismus   MeSH
• zdraví dobrovolníci pro lékařské studie MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• kultivační média MeSH
• titan MeSH
• titanium dioxide MeSH Prohlížeč
• voda MeSH

UNLABELLED: Multiple myeloma plasma cells are actively dividing cells with the long surviving ability in the ex-vivo culture. In the effort for better understanding of the proliferative potential of malignant myeloma cells and establishment of permanent myeloma cell lines we performed long term cultures of human myeloma cells ex-vivo. During the last two years we cultured 41 bone marrow samples from 39 patients with multiple myeloma. Cells were cultured in the RPMI 1640 culture medium with 15% fetal calf serum at 37 degrees C in 5% CO2 and approximately one third of the culture medium was changed regularly twice a week. Most of the marrows cultures died by apoptosis within 30 days. Four bone marrow samples were cultured for more than 11 months, however, no culture can be qualified as an established cell line. In three cases permanent B-lymphoblastoid cell lines were established (UHKT-55, UHKT-56 a UHKT-57) but secondary immortalization by Epstein-Barr virus was suggested. CONCLUSIONS: Presented results suggest that myeloma plasma cells can survive and are able to proliferate in the ex-vivo culture for several months up to one year independently on the addition of any external growth factor without spontaneous apoptosis or necrosis. The probability of the establishment of a permanent cell line of plasma cell origin is, however, low. Presence of accessory bone marrow cells was the most important factor for the long-term survival of myeloma cells.

• MeSH
• buněčné kultury metody   MeSH
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• mnohočetný myelom patologie   MeSH
• nádorové buňky kultivované * MeSH
• plazmatické buňky patologie   MeSH
• senioři MeSH
• Check Tag
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• senioři MeSH
• Publikační typ
• anglický abstrakt MeSH
• časopisecké články MeSH
• práce podpořená grantem MeSH