ABSTRACT
MicroRNAs epigenetically regulate physiological and pathological processes. Previously, we found that miR-204-5p is expressed at low levels in melanoma cells, and an increase in its level leads to a change in proliferation, migration, and invasion of these cancer cells. Now, using bioinformatics analysis, it has been shown that the target of miR-204-5p is FOXC1 transcription factor, which is implicated in carcinogenesis. Using the luciferase reporter assay, it was found that miR-204-5p suppresses expression of the FOXC1 gene by binding to its 3' non-coding region. Transfection of small interfering RNA (siRNA) targeting FOXC1 into melanoma cells caused a decrease in miR-204-5p levels, which is consistent with the generally accepted concept of feedback regulation of miRNA expression by target genes. According to the results of the MTT test and fluorescence microscopy, the proliferation level of melanoma cells under the influence of siRNA to FOXC1 decreased 72 h after transfection. Changes in the ratio of cells by cell cycle phase were analyzed using flow cytometry. Regulatory relationships between FOXC1 and miR-204-5p, and an inhibitory effect of FOXC1 knockdown on melanoma cell proliferation were revealed. Based on the results, it can be assumed that miR-204-5p regulates proliferation of melanoma cells by affecting FOXC1 expression.
Subject(s)
Melanoma , MicroRNAs , Cell Line, Tumor , Cell Movement/genetics , Cell Proliferation/genetics , Forkhead Transcription Factors , Humans , Melanoma/genetics , MicroRNAs/geneticsABSTRACT
We studied changes in the subpopulation composition and phagocytic activity of monocytes in patients with kidney cancer under the influence of metabolites of the tumor microenvironment in vitro (lactate, ADP, and glutamate). Incubation with metabolites caused similar shifts in the monocyte subsets in kidney cancer patients (an increase in the relative content of classical CD14++CD16- monocytes and a decrease in the content of the fraction of intermediate CD14++CD16+ monocytes), but different changes in their phagocytic activity. The regulatory effect of metabolites on cells is realized by various mechanisms: receptor, metabolic, and epigenomic. The reactions of monocytes to metabolites in vitro confirm the existence of a distant metabolic effect of the tumor on blood cells that should be taken into account when developing new immunotherapeutic methods.
Subject(s)
Adenosine Diphosphate/pharmacology , Glutamic Acid/pharmacology , Kidney Neoplasms/immunology , Lactic Acid/pharmacology , Monocytes/drug effects , Phagocytosis/drug effects , Adult , Biomarkers/metabolism , Case-Control Studies , Cell Count , Female , Flow Cytometry , GPI-Linked Proteins/genetics , GPI-Linked Proteins/immunology , Gene Expression , Humans , Immunophenotyping , Kidney Neoplasms/genetics , Kidney Neoplasms/pathology , Lipopolysaccharide Receptors/genetics , Lipopolysaccharide Receptors/immunology , Male , Middle Aged , Monocytes/immunology , Monocytes/pathology , Primary Cell Culture , Receptors, IgG/genetics , Receptors, IgG/immunology , Tumor Microenvironment/immunologyABSTRACT
Adaptive immunity plays a crucial role in the system of anti-tumor defense of the organism. The theory of immune surveillance defines the presence of permanent monitoring of the organism for the detection of malignant transformed cells and their elimination or suppression of the growth. However in some cases tumor cells overcome immune surveillance that leads to the development of cancer.
