RESUMO
Experimental models that capture the genetic complexity of human disease and allow mechanistic explorations of the underlying cell, tissue, and organ interactions are crucial to furthering our understanding of disease biology. Such models require combinatorial manipulations of multiple genes, often in more than one tissue at once. The ability to perform complex genetic manipulations in vivo is a key strength of Drosophila, where many tools for sophisticated and orthogonal genetic perturbations exist. However, combining the large number of transgenes required to establish more representative disease models and conducting mechanistic studies in these already complex genetic backgrounds is challenging. Here we present a design that pushes the limits of Drosophila genetics by allowing targeted combinatorial ectopic expression and knockdown of multiple genes from a single inducible transgene. The polycistronic transcript encoded by this transgene includes a synthetic short hairpin cluster cloned within an intron placed at the 5' end of the transcript, followed by two protein-coding sequences separated by the T2A sequence that mediates ribosome skipping. This technology is particularly useful for modeling genetically complex diseases like cancer, which typically involve concurrent activation of multiple oncogenes and loss of multiple tumor suppressors. Furthermore, consolidating multiple genetic perturbations into a single transgene further streamlines the ability to perform combinatorial genetic manipulations and makes it readily adaptable to a broad palette of transgenic systems. This flexible design for combinatorial genetic perturbations will also be a valuable tool for functionally exploring multigenic gene signatures identified from omics studies of human disease and creating humanized Drosophila models to characterize disease-associated variants in human genes. It can also be adapted for studying biological processes underlying normal tissue homeostasis and development that require simultaneous manipulation of many genes.
Assuntos
Drosophila , Técnicas Genéticas , Animais , Humanos , Drosophila/genética , Transgenes , Animais Geneticamente Modificados , ÍntronsRESUMO
MicroRNAs (miRNAs) are small, noncoding RNAs with important functions in development, cell differentiation, and regulation of cell cycle and apoptosis. MiRNA expression is deregulated in various pathological processes including tumorigenesis and cancer progression through various mechanisms including amplification or deletion of miRNA genes, mutations, and epigenetic silencing and defects in the miRNA biogenesis machinery. Several studies have now shown abnormal miRNA profiles and proved their involvement in the initiation and progression of cancer. Since miR-452 has diverse roles (as suppressor or oncogene) in different cellular processes including epithelial-mesenchymal transition (EMT), proliferation, migration, and invasion, in this review we highlight a brief overview of the biological function and regulatory mechanism of miR-452 and its involvement as a potential biomarker for diagnosis and treatment of various cancer types.
Assuntos
MicroRNAs , Neoplasias , Humanos , Carcinogênese/genética , Linhagem Celular Tumoral , Proliferação de Células/genética , Transformação Celular Neoplásica/genética , Transição Epitelial-Mesenquimal/genética , Regulação Neoplásica da Expressão Gênica , MicroRNAs/genética , MicroRNAs/metabolismo , Neoplasias/genética , Neoplasias/patologia , OncogenesRESUMO
Nucleophosmin (NPM1) is a well-known nucleocytoplasmic shuttling protein that performs several cellular functions such as ribosome biogenesis, chromatin remodeling, genomic stability, cell cycle progression, and apoptosis. NPM1 has been identified to be necessary for normal cellular functions, and its altered regulation by overexpression, mutation, translocation, loss of function, or sporadic deletion can lead to cancer and tumorigenesis. In this review, we focus on the gene and protein structure of NPM1 and its physiological roles. Finally, we discuss the association of NPM1 with various types of cancer including solid tumors and leukemia.
Assuntos
Biomarcadores Tumorais/metabolismo , Transformação Celular Neoplásica/metabolismo , Neoplasias/metabolismo , Nucleofosmina/metabolismo , Animais , Biomarcadores Tumorais/genética , Transformação Celular Neoplásica/genética , Transformação Celular Neoplásica/patologia , Progressão da Doença , Regulação Neoplásica da Expressão Gênica , Humanos , Mutação , Neoplasias/genética , Neoplasias/patologia , Nucleofosmina/genética , Transdução de SinaisRESUMO
Colorectal cancer (CRC) is the most frequently diagnosed cancer and the most common gastrointestinal cancer worldwide. Due to the presence of populations of cancer stem cells (CSCs) that cause recurrence, the possibility of cancer treatment is very low. The aim of current study was to evaluate the inhibitory role of miR-200c on EMT, CSCs markers and ß-catenin in HCT-116 and SW48 cell lines. The expression of miR-200c, EMT-related genes, CSCs markers, and ß-catenin were quantified by qRT-PCR. Further, expression of ß-catenin and EMT-related proteins, and migration were analyzed by Western blot, and migration assay kit, respectively. Spheroid formation assay was used to enrich colorectal CSCs from colorectal cancer cell lines. LNA-anti-miR-200c suppressed the endogenous miR-200c in transfected cells compared with the control. qRT PCR and Western blot analysis of LNA-anti-miR-200c transfected cells revealed a considerable increase in CSCs markers, vimentin, ZEB-1, N-cadherin, and ß-catenin expression, with a concomitant reduction in E-cadherin expression level. Migration and sphere forming ability of HCT-116 and SW48 cells increased in transfected cells. The results of current study revealed that downregulation of miR-200c may be an important factor for the overexpression of CSCs markers and EMT related genes via ß-catenin upregulation in CRC.
