elektronický časopis
Sunitinib malate is a small molecule that targets multiple receptor tyrosine kinases and blocks their activity. Receptors targeted by sunitinib are implicated in tumor vascularization and are overexpressed by vascular tumors encountered in infants, namely, hemangiomas. Of note is that there is still no definitive treatment for these commonly occurring tumors of infancy. The purpose of this study was to investigate the effects of sunitinib malate on hemangioma using endothelial cells isolated from a murine model of the neoplasm (sEnd.2). The effects of the drug on cell growth were evaluated using the crystal violet assay and flow cytometry, while the scratch assay was employed to measure cell migration. Proteins associated with cell migration and angiogenesis were detected using western blotting. Sunitinib was investigated further to determine its effects on the production of reactive oxygen species, a parameter associated with the promotion of neovascularization in tumors. The results showed that sunitinib significantly reduced the growth of sEnd.2 cells by causing the cells to accumulate in the sub-G1 phase of the cell cycle, and also induced a significant decrease in the migration of these hemangioma cells (P < 0.05). The western blot assay showed a decrease in the expression of adhesion proteins, focal adhesion kinase and paxillin at IC50 doses, although the expression of cadherin did not change significantly (P < 0.05). In addition, transforming growth factor-β1 (TGF-β1) expression was decreased in sunitinib-treated cells at the same dose. The adhesion proteins as well as TGF-β1 regulate cell movement and have been implicated in tumor progression. Thus, sunitinib malate may have potential in the treatment of hemangiomas.
- MeSH
- Cell Migration Assays statistics & numerical data MeSH
- Cell Cycle MeSH
- Cell Growth Processes drug effects MeSH
- Endothelial Cells drug effects MeSH
- Focal Adhesion Protein-Tyrosine Kinases MeSH
- Hemangioma * drug therapy MeSH
- Models, Animal MeSH
- Mice MeSH
- Flow Cytometry MeSH
- Statistics as Topic MeSH
- Sunitinib * pharmacology therapeutic use MeSH
- Vascular Endothelial Growth Factors MeSH
- Cell Survival MeSH
- Treatment Outcome MeSH
- Blotting, Western MeSH
- Check Tag
- Mice MeSH
- Publication type
- Clinical Study MeSH
- Research Support, Non-U.S. Gov't MeSH
Cell fate modulation by adapting the surface of a biocompatible material is nowadays a challenge in implantology, tissue engineering as well as in construction of biosensors. Nanocrystalline diamond (NCD) thin films are considered promising in these fields due to their extraordinary physical and chemical properties and diverse ways in which they can be modified structurally and chemically. The initial cell distribution, the rate of cell adhesion, distance of cell migration and also the cell proliferation are influenced by the NCD surface termination. Here, we use real-time live-cell imaging to investigate the above-mentioned processes on oxidized NCD (NCD-O) and hydrogenated NCD (NCD-H) to elucidate cell preference to the NCD-O especially on surfaces with microscopic surface termination patterns. Cells adhere more slowly and migrate farther on NCD-H than on NCD-O. Cells seeded with a fetal bovine serum (FBS) supplement in the medium move across the surface prior to adhesion. In the absence of FBS, the cells adhere immediately, but still exhibit different migration and proliferation on NCD-O/H regions. We discuss the impact of these effects on the formation of cell arrays on micropatterned NCD. © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 105A: 1469-1478, 2017.
Epithelial ovarian cancer (EOC) cells express Wnt5a, but its role in ovarian cancer progression is poorly defined. The aims of the present study were two-fold: 1) to determine the Wnt5a role in viability, apoptosis, migration, colony formation and adhesion of human serous epithelial ovarian cancer cell line SKOV-3, and 2) to assess the relationship of Wnt5a with E- and N-cadherin in high- and low-grade human serous ovarian cancer specimens. Wnt5a over-expression led to 29% increased serum-independent cell viability (P < 0.05) and 35% decreased caspase-3 activity (P < 0.01) compared to SKOV-3 cells. There was 96% (P < 0.001) increased cell motility in Wnt5a-transfected SKOV-3 (SKOV-3/Wnt5a) cells compared to SKOV-3, which was abrogated in the presence of JNK inhibitor. In addition, there was about 42% increased cell adhesion to Matrigel compared to SKOV-3 cells (P < 0.001). Colony-forming assay showed a 4.4-fold increased colony formation in SKOV-3/Wnt5a cells compared to SKOV-3 cells (P < 0.001). E- and N-cadherin levels were reduced by 49 % and 67 % in SKOV-3/Wnt5a cells compared to mock cells, respectively. Wnt5a and E-cadherin immunoexpression was significantly (P < 0.001) different in low-grade serous ovarian cancer (LGSC) and high-grade serous ovarian cancer (HGSC). In HGSC specimens, strong immunoexpression of Wnt5a was detected compared to LGSC. However, E-cadherin showed moderate immunostaining (84 %) in HGSC, whereas 100 % of LGSC specimens showed strong immunoexpression. In both groups no N-cadherin immunoexpression was detected. Moreover, Wnt5a showed a positive relationship with E-cadherin in the LGSC group (r = 0.661, P = 0.027). These results may support important roles for Wnt5a in EOC progression.
- MeSH
- Cell Adhesion genetics physiology MeSH
- Cadherins genetics metabolism MeSH
- Humans MeSH
- Cell Line, Tumor MeSH
- Neoplasms, Glandular and Epithelial genetics metabolism MeSH
- Ovarian Neoplasms genetics metabolism MeSH
- Cell Movement genetics physiology MeSH
- Cell Proliferation MeSH
- Wnt Proteins genetics metabolism MeSH
- Proto-Oncogene Proteins genetics metabolism MeSH
- Cell Survival genetics physiology MeSH
- Check Tag
- Humans MeSH
- Female MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
Surface protein CD20 serves as the critical target of immunotherapy in various B-cell malignancies for decades, however its biological function and regulation remain largely elusive. Better understanding of CD20 function may help to design improved rational therapies to prevent development of resistance. Using CRISPR/Cas9 technique, we have abrogated CD20 expression in five different malignant B-cell lines. We show that CD20 deletion has no effect upon B-cell receptor signaling or calcium flux. Also B-cell survival and proliferation is unaffected in the absence of CD20. On the contrary, we found a strong defect in actin cytoskeleton polymerization and, consequently, defective cell adhesion and migration in response to homeostatic chemokines SDF1α, CCL19 and CCL21. Mechanistically, we could identify a reduction in chemokine-triggered PYK2 activation, a calcium-activated signaling protein involved in activation of MAP kinases and cytoskeleton regulation. These cellular defects in consequence result in a severely disturbed homing of B cells in vivo.
- MeSH
- Actins metabolism MeSH
- Antigens, CD20 genetics metabolism physiology MeSH
- Lymphoma, B-Cell metabolism pathology MeSH
- B-Lymphocytes pathology physiology MeSH
- Cell Adhesion physiology MeSH
- Gene Knockdown Techniques MeSH
- Leukemia, B-Cell metabolism pathology MeSH
- Humans MeSH
- Protein Multimerization physiology MeSH
- Mice, Inbred NOD MeSH
- Mice, SCID MeSH
- Mice, Transgenic MeSH
- Mice MeSH
- Cell Line, Tumor MeSH
- Cell Movement physiology MeSH
- Polymerization MeSH
- Receptors, Antigen, B-Cell metabolism MeSH
- Signal Transduction immunology MeSH
- Animals MeSH
- Check Tag
- Humans MeSH
- Mice MeSH
- Animals MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
Triple-negative breast cancers (TNBC) comprise a heterogeneous subgroup of tumors with a generally poor prognosis. Subclassification of TNBC based on genomic analyses shows that basal-like TNBCs, specifically the basal A or BL2 subtype, are characterized by the expression of ΔNp63, a transcription factor that has been attributed a variety of roles in the regulation of proliferation, differentiation, and cell survival. To investigate the role(s) of p63 in basal-like breast cancers, we used HCC1806 cells that are classified as basal A/BL2. We show that these cells endogenously express p63, mainly as the ΔNp63α isoform. TP63 gene knockout by CRISPR resulted in viable cells that proliferate more slowly and adhere less tightly, with an increased rate of migration. Analysis of adhesion-related gene expression revealed a complex set of alterations in p63-depleted cells, with both increased and decreased adhesion molecules and adhesion substrates compared to parental cells expressing p63. Examination of the phenotype of these cells indicated that endogenous p63 is required to suppress the expression of luminal markers and maintain the basal epithelial phenotype, with increased levels of both CK8 and CK18 and a reduction in N-cadherin levels in cells lacking p63. On the other hand, the level of CK5 was not decreased and ER was not increased, indicating that p63 loss is insufficient to induce full luminal-type differentiation. Taken together, these data demonstrate that p63 exerts multiple pro-oncogenic effects on cell differentiation, proliferation and adhesion in basal-like breast cancers.
- MeSH
- Cell Adhesion MeSH
- Cell Differentiation MeSH
- Antigens, CD biosynthesis genetics MeSH
- CRISPR-Cas Systems MeSH
- Epithelial Cells metabolism pathology MeSH
- Phenotype MeSH
- Gene Knockout Techniques MeSH
- Cadherins biosynthesis genetics MeSH
- Carcinoma pathology MeSH
- Keratins biosynthesis genetics MeSH
- Humans MeSH
- Cell Line, Tumor MeSH
- Neoplasm Proteins physiology MeSH
- Tumor Suppressor Proteins deficiency physiology MeSH
- Cell Proliferation MeSH
- Protein Isoforms physiology MeSH
- Gene Expression Regulation, Neoplastic MeSH
- Transcription Factors deficiency physiology MeSH
- Triple Negative Breast Neoplasms pathology MeSH
- Check Tag
- Humans MeSH
- Female MeSH
- Publication type
- Journal Article MeSH
INTRODUCTION: Breast cancer is one of the most common types of cancer in women. One of the genes that were found mutated in breast cancer is casein kinase 1 epsilon (CK1epsilon). Because CK1epsilon is a crucial regulator of the Wnt signaling cascades, we determined how these CK1epsilon mutations interfere with the Wnt pathway and affect the behavior of epithelial breast cancer cell lines. METHODS: We performed in silico modeling of various mutations and analyzed the kinase activity of the CK1epsilon mutants both in vitro and in vivo. Furthermore, we used reporter and small GTPase assays to identify how mutation of CK1epsilon affects different branches of the Wnt signaling pathway. Based on these results, we employed cell adhesion and cell migration assays in MCF7 cells to demonstrate a crucial role for CK1epsilon in these processes. RESULTS: In silico modeling and in vivo data showed that autophosphorylation at Thr 44, a site adjacent to the breast cancer point mutations in the N-terminal lobe of human CK1epsilon, is involved in positive regulation of the CK1epsilon activity. Our data further demonstrate that, in mammalian cells, mutated forms of CK1epsilon failed to affect the intracellular localization and phosphorylation of Dvl2; we were able to demonstrate that CK1epsilon mutants were unable to enhance Dvl-induced TCF/LEF-mediated transcription, that CK1epsilon mutants acted as loss-of-function in the Wnt/beta-catenin pathway, and that CK1epsilon mutants activated the noncanonical Wnt/Rac-1 and NFAT pathways, similar to pharmacological inhibitors of CK1. In line with these findings, inhibition of CK1 promoted cell migration as well as decreased cell adhesion and E-cadherin expression in the breast cancer-derived cell line MCF7. CONCLUSIONS: In summary, these data suggest that the mutations of CK1epsilon found in breast cancer can suppress Wnt/beta-catenin as well as promote the Wnt/Rac-1/JNK and Wnt/NFAT pathways, thus contributing to breast cancer development via effects on cell adhesion and migration. In terms of molecular mechanism, our data indicate that the breast cancer point mutations in the N-terminal lobe of CK1epsilon, which are correlated with decreased phosphorylation activities of mutated forms of CK1epsilon both in vitro and in vivo, interfere with positive autophosphorylation at Thr 44.
- MeSH
- beta Catenin metabolism MeSH
- Cell Adhesion MeSH
- Carcinoma, Ductal, Breast genetics metabolism pathology MeSH
- Phosphorylation MeSH
- Immunoprecipitation MeSH
- Casein Kinase 1 epsilon chemistry genetics metabolism MeSH
- Protein Conformation MeSH
- Humans MeSH
- MAP Kinase Kinase 4 metabolism MeSH
- RNA, Messenger genetics MeSH
- Mutation genetics MeSH
- Cell Line, Tumor MeSH
- Breast Neoplasms genetics metabolism pathology MeSH
- Cell Movement MeSH
- Reverse Transcriptase Polymerase Chain Reaction MeSH
- Cell Proliferation MeSH
- Wnt Proteins metabolism MeSH
- rac1 GTP-Binding Protein metabolism MeSH
- NFATC Transcription Factors metabolism MeSH
- Blotting, Western MeSH
- Check Tag
- Humans MeSH
- Female MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
- Research Support, N.I.H., Extramural MeSH
Proper course of folliculogenesis and oogenesis have an enormous impact on female fertility. Both processes take place in the ovary and involve not only the maturing germ cell, but also few types of somatic cells that assist the ovarian processes and mediate the dialog with the oocyte. These cells, granulosa and theca, are heavily involved in essential reproductive processes, such as ovulation, fertilization, and embryo implantation. In this study, we have used the expressive microarray approach to analyze the transcriptome of porcine granulosa cells, during short-term in vitro culture. We have further selected differentially expressed gene ontologies, involved in cell proliferation, migration, adhesion, and tissue development, namely, "cell-cell adhesion," "cell motility," "cell proliferation," "tissue development," and "tissue migration" to screen them for the possibility of discovery of new markers of those processes. A total of 303 genes, expression of which varied significantly in different culture periods and belonged to the analyzed ontology groups, were detected, of which 15 that varied the most (between 0 and 48 h of culture) were selected for validation. As the validation confirmed the transcriptomic patterns, 10 genes of biggest changes in expression (CAV1, IGFBP5, ITGB3, FN1, ITGA2, LAMB1, POSTN, FAM83D, KIF14, and CDK1) were analyzed, described, and referred to the context of the study, with the most promising new markers and further proof for the viability of the currently recognized ones detailed. Overall, the study provided valuable insight into the molecular functioning of in vitro granulosa cell cultures.
- MeSH
- Biomarkers metabolism MeSH
- Cell Adhesion genetics MeSH
- Granulosa Cells cytology metabolism physiology MeSH
- Cells, Cultured MeSH
- Cell Movement genetics MeSH
- Swine MeSH
- Cell Proliferation genetics MeSH
- Oligonucleotide Array Sequence Analysis MeSH
- Gene Expression Profiling MeSH
- Animals MeSH
- Check Tag
- Female MeSH
- Animals MeSH
- Publication type
- Journal Article MeSH
Polyaniline shows great potential and promises wide application in the biomedical field thanks to its intrinsic conductivity and material properties, which closely resemble natural tissues. Surface properties are crucial, as these predetermine any interaction with biological fluids, proteins and cells. An advantage of polyaniline is the simple modification of its surface, e.g., by using various dopant acids. An investigation was made into the adhesion, proliferation and migration of mouse embryonic fibroblasts on pristine polyaniline films and films doped with sulfamic and phosphotungstic acids. In addition, polyaniline films supplemented with poly (2-acrylamido-2-methyl-1-propanesulfonic) acid at various ratios were tested. Results showed that the NIH/3T3 cell line was able to adhere, proliferate and migrate on the pristine polyaniline films as well as those films doped with sulfamic and phosphotungstic acids; thus, utilization of said forms in biomedicine appears promising. Nevertheless, incorporating poly (2-acrylamido-2-methyl-1-propanesulfonic) acid altered the surface properties of the polyaniline films and significantly affected cell behavior. In order to reveal the crucial factor influencing the surface/cell interaction, cell behavior is discussed in the context of the surface energy of individual samples. It was clearly demonstrated that the lesser the difference between the surface energy of the sample and cell, the more cyto-compatible the surface is.
- MeSH
- Aniline Compounds chemistry pharmacology MeSH
- Biocompatible Materials chemistry pharmacology MeSH
- Cell Adhesion drug effects MeSH
- NIH 3T3 Cells MeSH
- Mice MeSH
- Cell Movement drug effects MeSH
- Surface Properties MeSH
- Cell Proliferation drug effects MeSH
- Animals MeSH
- Check Tag
- Mice MeSH
- Animals MeSH
- Publication type
- Journal Article MeSH
BACKGROUND: Colon cancer is the most common type of gastrointestinal cancer. Despite advances during the last two decades, the efficacy of colorectal cancer treatment is still insufficient and new anticancer agents are necessary. METHODS: In our study, colon cancer cells derived from a primary tumor (SW480) and lymph node metastasis (SW620) from the same patient were used and compared. The effect of flubendazole (FLU) on cell adhesion and migration was monitored using the x-CELLigence Real-Time Cell Analysis system. Expressions of molecules involved in adhesion and migration were analyzed using RT-PCR and western blot. Furthermore, RNA silencing of nuclear factor-κB in SW620 cells was used to determine the involvement of the NF-κB p65 regulation pathway in FLU action. RESULTS: FLU significantly suppressed the adhesion of SW480 cells and reduced the expression of adhesion markers (ICAM-1, αE-catenin; β-catenin; integrin α5 and β1). Moreover, a significant anti-migratory potential of FLU was manifested in the SW620 cells. In addition, FLU suppressed the phosphorylation of NF-κB p65 and potentiated the suppression of several metastatic markers (ICAM-1, EpCAM, integrin α5, β1, α-tubulin) caused by NF-κB p65 silencing. CONCLUSION: FLU has a significant anti-migratory effect in intestinal cancer cell SW480 and its lymph node metastatic cells SW620. FLU decreases the expression of some proteins involved in metastatic processes and inhibits activation of NF-κB p65.
- MeSH
- Cell Adhesion drug effects MeSH
- Humans MeSH
- Mebendazole analogs & derivatives chemistry pharmacology MeSH
- Molecular Structure MeSH
- Tumor Cells, Cultured MeSH
- Colonic Neoplasms drug therapy pathology MeSH
- Cell Movement drug effects MeSH
- Cell Proliferation drug effects MeSH
- Antineoplastic Agents chemistry pharmacology MeSH
- Drug Screening Assays, Antitumor MeSH
- Dose-Response Relationship, Drug MeSH
- Structure-Activity Relationship MeSH
- Check Tag
- Humans MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
