RESUMEN
Quantitative real time reverse transcription PCR, qRT-PCR, is one of the most important techniques for assessing the level of gene expression. Selecting the correct reference gene to normalize the results is a key step in this method. Inaccurate data can be generated if the correct reference gene is not selected. The level of the expression of reference genes is tissue-variable, and in the case of mesenchymal stem cells (MSC), it can be different depending on the source of their origin. The aim of this study was to select the reference gene for Wharton's Jelly-derived MSC (WJ- MSC) that were undergoing transduction and differentiation. In this work, the expression of 32 genes was analyzed, of which two (RPS17 and 18S rRNA), which had the most stable expression level, were selected. A comparative analysis of the expression stability of the selected genes was then performed with the genes that are most commonly used in the literature, i.e. ß-actin and GAPDH. Next, it was determined that a false picture of the expression level of the studied genes can be obtained when a reference gene with variable expression level is used for normalization. RPS17 and 18S rRNA proved to be the most stable reference genes for the WJ-MSC that had been subjected to the lentiviral transfection procedure followed by differentiation. The expression of ß-actin and GAPDH was highly unstable and therefore these genes are not suitable for use as reference genes in studies involving WJ- MSC.
Asunto(s)
Diferenciación Celular/genética , Vectores Genéticos/genética , Lentivirus/genética , Células Madre Mesenquimatosas/citología , Células Madre Mesenquimatosas/metabolismo , Transducción Genética , Gelatina de Wharton/citología , Adipogénesis , Biomarcadores , Células Cultivadas , Humanos , Inmunofenotipificación , Osteogénesis , TransgenesRESUMEN
A recurrent proximal microdeletion at 15q25.2 with an approximate 1.5 megabase smallest region of overlap has recently been reported in seven patients and is proposed to be associated with congenital diaphragmatic hernia (CDH), mild to moderate cognitive deficit, and/or features consistent with Diamond-Blackfan anemia. We report on four further patients and define the core phenotypic features of individuals with this microdeletion to include mild to moderate developmental delay or intellectual disability, postnatal short stature, anemia, and cryptorchidism in males. CDH and structural organ malformations appear to be less frequent associations, as is venous thrombosis. There is no consistent facial dysmorphism. Features novel to our patient group include dextrocardia, obstructive sleep apnea, and cleft lip.
Asunto(s)
Deleción Cromosómica , Cromosomas Humanos Par 15 , Fenotipo , Anomalías Múltiples/diagnóstico , Anomalías Múltiples/genética , Adolescente , Niño , Preescolar , Mapeo Cromosómico , Hibridación Genómica Comparativa , Femenino , Heterocigoto , Humanos , Lactante , Masculino , Polimorfismo de Nucleótido Simple , SíndromeRESUMEN
The prognosis for patients with colorectal cancer (CRC) remains worse than expected due to metastasis, recurrence, and resistance to chemotherapy. Colorectal cancer stem cells (CRCSCs) play a vital role in tumor metastasis, recurrence, and chemotherapy resistance. However, there are currently no prognostic markers based on CRCSCs-related genes available for clinical use. In this study, single-cell transcriptome sequencing was employed to distinguish cancer stem cells (CSCs) in the CRC microenvironment and analyze their properties at the single-cell level. Subsequently, data from TCGA and GEO databases were utilized to develop a prognostic risk model for CRCSCs-related genes and validate its diagnostic performance. Additionally, functional enrichment, immune response, and chemotherapeutic drug sensitivity of the relevant genes in the risk model were investigated. Lastly, the key gene RPS17 in the risk model was identified as a potential prognostic marker and therapeutic target for further comprehensive studies. Our findings provide new insights into the prognostic treatment of CRC and offer novel perspectives for a systematic and comprehensive understanding of CRC development.
Asunto(s)
Biomarcadores de Tumor , Neoplasias Colorrectales , Células Madre Neoplásicas , RNA-Seq , Análisis de la Célula Individual , Humanos , Neoplasias Colorrectales/genética , Neoplasias Colorrectales/patología , Neoplasias Colorrectales/mortalidad , Células Madre Neoplásicas/metabolismo , Células Madre Neoplásicas/patología , Análisis de la Célula Individual/métodos , Pronóstico , Biomarcadores de Tumor/genética , Regulación Neoplásica de la Expresión Génica , Microambiente Tumoral/genética , Transcriptoma , Perfilación de la Expresión Génica , Análisis de Secuencia de ARN/métodosRESUMEN
The aim of this study was to analyze the expression patterns of key genes involved in lipid metabolism in response to feeding in chicks. A total of 18 thirteen day-old male chicks were fasted for 12h. The mRNA levels of the genes in the liver and white adipose tissue were analyzed after 0, 2, and 4h of refeeding. The mRNA levels of sterol regulatory element-binding protein (SREBP) 1, liver X receptor α, peroxisome proliferator-activated receptor (PPAR) γ, acetyl-CoA carboxylase α and fatty acid synthase were significantly increased after 2h of refeeding. In contrast, the mRNA levels of PPARα and carnitine palmitoyltransferase 1a were significantly decreased after 2h of refeeding. The mRNA level of acyl-CoA oxidase was significantly decreased after 4h of refeeding. The mRNA levels of cholesterol metabolism-related genes such as SREBP2 and 3-hydroxy-3-methylglutaryl-CoA reductase were significantly increased after 2h of refeeding. In the white adipose tissue, the mRNA level of PPARγ was significantly increased after 2h of refeeding, whereas the mRNA level of adipose triglyceride lipase was significantly decreased after 4h of refeeding. These results demonstrated that expression of lipid metabolism-related genes is regulated by short-term refeeding in chicks.