ABSTRACT
Kabuki syndrome (KS) is a rare genetic disease that causes developmental delay and congenital anomalies. Since the identification of MLL2 mutations as the primary cause of KS, such mutations have been identified in 56%-76% of affected individuals, suggesting that there may be additional genes associated with KS. Here, we describe three KS individuals with de novo partial or complete deletions of an X chromosome gene, KDM6A, that encodes a histone demethylase that interacts with MLL2. Although KDM6A escapes X inactivation, we found a skewed X inactivation pattern, in which the deleted X chromosome was inactivated in the majority of the cells. This study identifies KDM6A mutations as another cause of KS and highlights the growing role of histone methylases and histone demethylases in multiple-congenital-anomaly and intellectual-disability syndromes.
Subject(s)
Abnormalities, Multiple/genetics , DNA-Binding Proteins/metabolism , Gene Deletion , Hematologic Diseases/genetics , Histone Demethylases/genetics , Neoplasm Proteins/metabolism , Nuclear Proteins/genetics , Vestibular Diseases/genetics , Abnormalities, Multiple/metabolism , Adolescent , Base Sequence , Child, Preschool , Chromosomes, Human, X/genetics , Developmental Disabilities/genetics , Face/abnormalities , Female , Hematologic Diseases/metabolism , Histone Demethylases/metabolism , Humans , Infant , Intellectual Disability/genetics , Male , Molecular Sequence Data , Nuclear Proteins/metabolism , Vestibular Diseases/metabolismABSTRACT
Surface properties of nanoparticles to be used for radioimmunotherapy need to be optimized to allow antibody conjugation while ensuring biocompatibility. We aimed to investigate cell adhesion and proliferation onto different coatings to be used for nanoparticles. C, CH(x) or SiO(x) coatings deposited onto glass coverslips by magnetron deposition as well as nitrogen functionalized materials synthetized using different reactive sputtering conditions and PPAA (plasma polymerized allylamine) coating, were compared. Amine functionalization did increase hydrophilicity in all the materials tested. Biocompatibility was assessed by measuring cell viability, morphology, attachment, spreading, and pro-inflammatory cytokine secretion. The results show that C and CN(x) were the most biocompatible substrates while SiO(x) and SiO(x)N(y) were the most toxic materials. PPAA coatings displayed unexpectedly an intermediate biocompatibility. A correlation could be observed between wettability and cell proliferation except for C coated surface, indicating that more complex processes than hydrophilicity alone are taking place that affect cell functions.
Subject(s)
Allylamine/chemistry , Amines/chemistry , Antibodies/chemistry , Biocompatible Materials/chemistry , Cell Adhesion , Cell Survival , Coated Materials, Biocompatible/chemistry , Cytokines/metabolism , Endothelial Cells/cytology , Glass , Humans , Inflammation , Nanoparticles/chemistry , Nanotechnology/methods , Radioimmunotherapy/methods , Surface PropertiesABSTRACT
BACKGROUND: it is now well established that hypoxia renders tumor cells resistant to radio- but also chemotherapy. However, few elements are currently available as for the mechanisms underlying this protection. RESULTS: in this study, physiological hypoxia was shown to inhibit apoptosis induced in HepG2 cells by etoposide. Indeed, hypoxia reduced DNA fragmentation, caspase activation and PARP cleavage. The DNA binding activity of 10 transcription factors was followed while the actual transcriptional activity was measured using specific reporter plasmids. Of note is the inhibition of the etoposide-induced activation of p53 under hypoxia. In parallel, data from low density DNA microarrays indicate that the expression of several pro- and anti-apoptotic genes was modified, among which are Bax and Bak whose expression profile paralleled p53 activity. Cluster analysis of data unravels several possible pathways involved in the hypoxia-induced protection against etoposide-induced apoptosis: one of them could be the inhibition of p53 activity under hypoxia since caspase 3 activity parallels Bax and Bak expression profile. Moreover, specific downregulation of HIF-1alpha by RNA interference significantly enhanced apoptosis under hypoxia possibly by preventing the hypoxia mediated decrease in Bak expression without altering Bax expression. CONCLUSION: these results are a clear demonstration that hypoxia has a direct protective effect on apoptotic cell death. Moreover, molecular profiling points to putative pathways responsible for tumor growth in challenging environmental conditions and cancer cell resistance to chemotherapeutic agents.
Subject(s)
Apoptosis/drug effects , Cytoprotection/drug effects , Etoposide/pharmacology , Gene Expression Profiling , Gene Expression Regulation, Neoplastic/drug effects , Transcription Factors/metabolism , Cell Hypoxia/drug effects , Cell Line, Tumor , Cluster Analysis , DNA, Neoplasm/metabolism , Gene Silencing , Genes, Reporter , Humans , Hypoxia-Inducible Factor 1, alpha Subunit/genetics , Oligonucleotide Array Sequence Analysis , RNA, Small Interfering/metabolism , Reverse Transcriptase Polymerase Chain Reaction , TransfectionABSTRACT
The Myc oncogene is deregulated in a wide variety of human tumors hence the Myc pathway is an attractive target for tailored cancer treatment. We have recently identified two small molecules, MYRAs (Myc-pathway response agents), that induce apoptosis in a Myc-dependent manner and inhibit Myc-driven transformation. Here, we show that these compounds in addition have prominent effects in MYCN overexpressing neuroblastoma cells. A third compound, NSC308848, also induced apoptosis in Myc-overexpressing cells and inhibited Myc-induced cellular transformation. However, in contrast to the MYRAs, NSC308848 treatment resulted in decreased Myc protein levels and gave rise to inhibitory effects also on other transcription factors than Myc. Taken together, our findings suggest that these three small molecules can elicit a similar biological response by interfering with the Myc pathway at different levels.
