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1.
Biochem Biophys Rep ; 38: 101715, 2024 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-38698835

RESUMEN

Nicotinamide adenine dinucleotide (NAD+) is an essential coenzyme involved in many pathophysiological processes. Supplementation with NAD+ and its precursors has been demonstrated as an emerging therapeutic strategy for the diseases. NAD+ also plays an important role in the reproductive system. Here, we summarize the function of NAD+ in various reproductive diseases and review the application of NAD+ and its precursors in the preservation of reproductive capacity and the prevention of embryonic malformations. It is shown that NAD+ shows good promise as a therapeutic approach for saving reproductive capacity.

2.
Cell Biochem Biophys ; 81(4): 727-735, 2023 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-37658975

RESUMEN

NLRP12 can affect the progression of different diseases, including hepatocellular carcinoma. However, no report on triple-negative breast cancer (TNBC) has been found. Thus, this study aimed to explore the role of NLRP12 in TNBC. In our study, immunohistochemistry, real-time quantitative PCR (qPCR), and Western blot assays were used to evaluate NLRP12 expression in TNBC tissues and cells. Then, NLRP12 lentivirus was constructed and infected into MDA-MB-231 and MDA-MB-157 cells with or without PTD-p65-P1 treatment. Next, cells were collected for cell function detection using the following procedures: colony formation assay for proliferation, Transwell for migration and invasion, and Western blot for NF-κB and MAPK pathway-associated proteins. Finally, a xenograft mouse model was applied; the tumor volume and weight were determined, and NLRP12, p-IκBb-α, and p-IκBb-α expressions were evaluated using qPCR and Western blot. Results indicated that NLRP12 was lowly expressed in TNBC tissues and cells. The inhibition of NLRP12 could induce the proliferation, migration, and invasion of TNBC cells, which also could be reversed by inhibiting the NF-κB pathway (PTD-p65-P1). Moreover, silencing of NLRP12 could upregulate p-IκBb-α, while IκBb-α, p-ERK, ERK, p-p38, p38, p-JNK, and JNK expressions remained unchanged, thereby indicating that only the NF-κB pathway could be activated by NLRP12 silencing. Furthermore, the xenograft mouse model confirmed the abovementioned findings. Therefore, the low expression of NLRP12 promoted the proliferation, migration, and invasion in TNBC cells by activating the NF-κB pathway. This study might provide insights into TNBC therapy.


Asunto(s)
FN-kappa B , Neoplasias de la Mama Triple Negativas , Humanos , Animales , Ratones , FN-kappa B/metabolismo , Transducción de Señal , Neoplasias de la Mama Triple Negativas/genética , Neoplasias de la Mama Triple Negativas/metabolismo , Neoplasias de la Mama Triple Negativas/patología , Línea Celular Tumoral , Modelos Animales de Enfermedad , Proliferación Celular , Movimiento Celular , Péptidos y Proteínas de Señalización Intracelular/metabolismo
3.
Org Lett ; 25(15): 2600-2605, 2023 Apr 21.
Artículo en Inglés | MEDLINE | ID: mdl-37017649

RESUMEN

Reported here is an efficient Friedel-Crafts-type reaction followed by electrocyclization cascade under an air atmosphere, employing readily available building blocks as starting materials, ethanol as a solvent, and Brønsted acid as a catalyst. On the basis of the cascade strategy, 2-(2-aminophenyl)-9H-carbazole has been furnished with excellent regioselectivity, exceptional functional group tolerance, and tolerated large-scale synthesis. Furthermore, one-pot syntheses of quino[3,4-a]carbazoles have been accomplished, demonstrating the broad synthetic utility of this strategy in the synthesis of valuable heteroaryl-annulated [a]carbazoles.

4.
Am J Med Genet A ; 191(5): 1240-1249, 2023 05.
Artículo en Inglés | MEDLINE | ID: mdl-36756859

RESUMEN

Coffin-Siris syndrome (CSS) 6 is caused by heterozygous pathogenic variants in the AT-rich interaction domain 2 (ARID2) gene on 12q12. Currently, only 26 cases with both detailed clinical and genetic information have been documented in the literature. Microdeletions of the entire ARID2 gene are rare. In this study, we report a 5-year-7-month-old Chinese female who underwent whole-exome sequencing to discover that she had a de novo 1.563 Mb heterozygous copy number loss at 12q12q13.11, involving an entire deletion of ARID2. The female had severe short stature with obvious dysmorphic facial features, global developmental delay and hypoplastic fingers and toes. Her growth hormone level was normal, with reduced IGF-1 and increased CA19-9 levels. After a review of the 27 patients with ARID2 deficiency, a significant positive correlation was observed between age and height standard deviation score (SDS) (r = 0.71, p = 0.0002), suggesting a possibility of growth catch-up. This study expands the genetic and phenotypic spectrum of CCS6 and provides a decision-making reference for growth hormone therapy.


Asunto(s)
Anomalías Múltiples , Enanismo , Deformidades Congénitas de la Mano , Discapacidad Intelectual , Micrognatismo , Femenino , Humanos , Lactante , Anomalías Múltiples/diagnóstico , Anomalías Múltiples/genética , Anomalías Múltiples/patología , Enanismo/genética , Cara/patología , Deformidades Congénitas de la Mano/diagnóstico , Deformidades Congénitas de la Mano/genética , Deformidades Congénitas de la Mano/patología , Discapacidad Intelectual/genética , Discapacidad Intelectual/patología , Micrognatismo/diagnóstico , Micrognatismo/genética , Micrognatismo/patología , Cuello/patología , Factores de Transcripción/genética
5.
Front Endocrinol (Lausanne) ; 13: 1015954, 2022.
Artículo en Inglés | MEDLINE | ID: mdl-36387899

RESUMEN

Objective: This study analyzed eight Chinese short stature children with aggrecan deficiency, and aimed to investigate potential genotype-phenotype correlations, differences in clinical characteristics between the Chinese and the Western populations, and effectiveness of recombinant human growth hormone therapy in patients with ACAN variants through a review of the literature. Methods: Pediatric short stature patients with ACAN heterozygous variants were identified using whole-exome sequencing. Subsequently, a literature review was carried out to summarize the clinical features, genetic findings, and efficacy of growth-promoting therapy in patients with ACAN variants. Results: We identified seven novel ACAN mutations and one recurrent variant. Patients in our center manifested with short stature (average height SDS: -3.30 ± 0.85) with slight dysmorphic characteristics. The prevalence of dysmorphic features in the Chinese populations is significantly lower than that in the Western populations. Meanwhile, only 24.24% of aggrecan-deficient Chinese children showed significantly advanced bone age (BA). Promising therapeutic benefits were seen in the patients who received growth-promoting treatment, with an increase in growth velocity from 4.52 ± 1.00 cm/year to 8.03 ± 1.16 cm/year. Conclusion: This study further expanded the variation spectrum of the ACAN gene and demonstrated that Chinese children with short stature who carried ACAN heterozygous variants exhibited early growth cessation, which may remain unnoticed by clinicians as most of these children had very mild dysmorphic characteristics and showed BA that was consistent with the chronological age. Genetic testing may help in the diagnosis.


Asunto(s)
Enanismo , Humanos , Niño , Agrecanos/genética , Heterocigoto , Enanismo/tratamiento farmacológico , Enanismo/genética , Pueblo Asiatico/genética , China/epidemiología
6.
Oncol Lett ; 21(6): 430, 2021 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-33868468

RESUMEN

Microsatellite instability (MSI) detection is widely used in the diagnosis and prognosis evaluation of colorectal cancer. However, for gastric cancer (GC), there is no standard panel of microsatellites (MSs) used in clinical guidance. The present study aimed to identify useful predictors of the clinical features and for the prognosis of GC, based on an investigation of MSI and loss of heterozygosity (LOH) in tumor-related genes. First, from 20 tumor-related genes which were proven to be important to the development of GC, 91 MSs were identified, and PCR amplification, short tandem repeat scanning analysis and TA clone sequencing were used to analyze MSI and LOH in the first set of 90 GC samples. Subsequently, the same method was used to detect the MSI/LOH of the optimized loci in the second set of 136 GC samples. MSI/LOH in the mismatch repair genes was highly consistent with that in oncogenes and tumor suppressor genes, respectively. The length of the core sequence was a main factor for the MSI/LOH rate. The MSI of 12 single loci was significantly associated with lymph node metastasis. The MSI in TP53-1 and the LOH in MGMT-10 were significantly associated with early stages of tumor infiltration depth. The LOH in MGMT-10, PTN-2 and MCC-17 was significantly associated with TNM stage. The LOH in TP53-1 and ERBB2-12 was associated with adenocarcinoma. The MSI/LOH in 6 single loci of 5 tumor-related genes was associated with poor prognosis of GC. The present study demonstrated that the MSI/LOH of loci in tumor-associated genes was associated with 4 clinicopathological characteristics and outcomes of GC. These results may provide potential specific biomarkers for the clinical prediction and treatment of GC.

7.
ACS Appl Mater Interfaces ; 13(8): 9932-9941, 2021 Mar 03.
Artículo en Inglés | MEDLINE | ID: mdl-33595272

RESUMEN

Rational design and controllable synthesis of multiple metal components according to chemical composition and morphology are essential for obtaining desirable electrochemical performance for efficient hydrogen production because of the morphology and synergistic effects of different components. Herein, we report an approach to facilely fabricate bimetal compounds with a well-defined hollow nanoprism structure using a self-templated strategy to synthesize novel hierarchical NiCo-layered double hydroxide (NiCo-LDH) nanosheets as precursors followed by in situ phosphorization. Among the as-synthesized products of different mole ratios of Ni/Co, the NiCo2-B-P nanoprisms that integrate the advantages of a hollow structure, an optimal Ni-Co synergistic effect, and a unique B-doped CoP/Ni2P bimetallic phosphide derived from NiCo-LDH nanosheets exhibit excellent hydrogen evolution reaction (HER) activity in an alkaline solution at 10 mA cm-2 with the lowest overpotential of 78 mV and long-term stability. This study may offer an appropriate structure and compositional design of bimetallic alkaline HER catalysts.

8.
Placenta ; 101: 115-123, 2020 11.
Artículo en Inglés | MEDLINE | ID: mdl-32950919

RESUMEN

Preeclampsia (PE) is the second most common complication that threatens the health of pregnant women and their foetuses; however, the underlying mechanisms are poorly understood. Circular RNAs (circRNAs) are involved in various human diseases, and an increasing number of studies have revealed the vital role of circRNAs in PE. Here, we investigated the biological function of circRNA-SFXN1 (CircSFXN1) in PE and the associated molecular mechanisms. Microarray data analysis revealed that CircSFXN1 was highly expressed in PE placentas compared to control placentas; this finding was confirmed by qRT-PCR. In vitro, CircSFXN1 overexpression significantly inhibited the invasion of TEV-1 trophoblasts and blocked the angiogenesis of human umbilical vein endothelial cells (HUVECs), while CircSFXN1 knockdown promoted trophoblast invasion and stimulated HUVEC angiogenesis. For in vivo evaluation, pregnant Sprague-Dawley rats were randomly selected for tail vein injection with sFLT1-expressing adenovirus, which resulted in elevated blood pressure and increased proteinuria; si-CircSFXN1 reversed these increases. Mechanistic analyses via RNA-protein pulldown, RNA immunoprecipitation (RIP) and dual-luciferase reporter assays showed that CircSFXN1 recruits sFLT1 and modulates the biological behaviour of trophoblasts by binding sFLT1. In summary, we identified a novel circRNA that regulates tumorigenic activities, suggesting a new pathway governing CircSFXN1/sFLT1-mediated regulation of trophoblast behaviour.


Asunto(s)
Preeclampsia/metabolismo , ARN Circular/fisiología , Transportador 1 de Sodio-Glucosa/genética , Trofoblastos/fisiología , Receptor 1 de Factores de Crecimiento Endotelial Vascular/metabolismo , Adulto , Animales , Estudios de Casos y Controles , Línea Celular , Femenino , Células Endoteliales de la Vena Umbilical Humana , Humanos , Neovascularización Fisiológica , Embarazo , Distribución Aleatoria , Ratas Sprague-Dawley
9.
RSC Adv ; 10(20): 12113-12118, 2020 Mar 19.
Artículo en Inglés | MEDLINE | ID: mdl-35496630

RESUMEN

Reported here is a novel cyclocondensation of aryl methyl ketones and triethyl orthoformate for the simple synthesis of m-terphenyls. In the presence of a catalytic amount of TfOH, alkyl- and chloro-substituted acetophenones produced a series of terphenyls through a tandem reaction which merged six steps into a one-pot procedure. Moreover, the corresponding ester products were obtained when using other substituted acetophenones as the starting materials under the same reaction conditions.

10.
Exp Ther Med ; 18(6): 4287-4294, 2019 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-31777536

RESUMEN

Microsatellite instability (MSI) and loss of heterozygosity (LOH), which cause genomic instability, contribute to cancer pathogenesis. However, only few studies have evaluated the association of a single microsatellite locus of the TP53 gene with the mutation spectra of TP53 exons. A total of 256 patients with colorectal cancer were enrolled in the present study. MSI/LOH alterations of a microsatellite in the TP53 intron (TP53ALU) were assessed via short tandem repeat scanning. The exon mutation profile was evaluated by direct sequencing. The mutation rate of TP53 exons was significantly higher in tumors with LOH alterations of TP53 introns compared with those in tumors with a microsatellite-stable status in the TP53 intron (P=0.0047). TNM stage II was significantly more frequent in MSI vs. LOH or MSS of the TP53 intron (P=0.027 and P=0.048, respectively). Thus, microsatellite alterations may be valuable predictors of TP53 exon mutation and the TNM stage of colorectal cancers.

11.
Phytomedicine ; 45: 36-40, 2018 Jun 01.
Artículo en Inglés | MEDLINE | ID: mdl-29550178

RESUMEN

BACKGROUND: Drugs derived from botany have been playing essential role in both clinical treatment and pharmaceutical industry, unfortunately our worry is still that its quality and therapeutic efficacy are inconsistent. Recently many scientists launched a new project on quality (Q)-marker of medicinal herbs, this study was thus designed to generate a novel concept of quality (Q)-markers: molecular connectivity index (MCI), and to test and verify the new concept of molecular connectivity index (MCI). METHODS: The first-order term (1χ) was selected to calculate and study quality (Q)-marker for TCM. Houttuynia cordata Thunb. (HCT) was adopted as a model to verify the hypothesis. Volatile oils of HCT were determined using gas chromatography-mass (GC-MS). SIMCA 13.0 and SPSS 21.0 were used to deal with the data. RESULTS: The minimum of the MCI values was 1.273, belonging to the peak 15, but the maximum (12.822) belonged to the peak 34, and the average value of fifty volatile oils was 5.798. The results demonstrated that MCI was the principle component, and monoterpenoid and sesquiterpenoid were also the principle components in oils. Fig. 2a shows peak 5, 24, 34 were the significant ingredients, while Fig. 2b shows peak 2, 5, 24 were the significant components. CONCLUSION: The data demonstrated that MCI was associated with the structure of molecules and the therapeutic efficacy, MCI could directly exhibit the relationship between ingredients and effectiveness of Traditional Chinese Medicine (TCM). So MCI could be a potential and promising parameter for quality (Q)-marker. Therefore, MCI may be developed as a novel potential concept to control the quality of TCM.


Asunto(s)
Biomarcadores Farmacológicos/análisis , Medicamentos Herbarios Chinos/química , Houttuynia/química , Aceites Volátiles/análisis , Biomarcadores Farmacológicos/química , Medicamentos Herbarios Chinos/farmacología , Medicamentos Herbarios Chinos/normas , Cromatografía de Gases y Espectrometría de Masas , Medicina Tradicional China/normas , Modelos Teóricos , Monoterpenos/análisis , Monoterpenos/química , Aceites Volátiles/química , Plantas Medicinales/química , Control de Calidad , Sesquiterpenos/análisis , Sesquiterpenos/química
12.
Biomed Pharmacother ; 96: 148-152, 2017 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-28972887

RESUMEN

Acute lung injury (ALI) and its severe form acute respiratory distress syndrome remain the leading cause of morbidity and mortality. LianQinJieDu (LQJD), which is a traditional Chinese medicine, has been clinically used for antiviral drug. The present study investigated whether Lianqinjiedu(LQJD) ameliorates lipopolysaccharide(LPS)-induced acute lung injury in rats and aimed to determine the anti-inflammatory effects of LQJD. Rat model with ALI induced by intraperitoneal injection of LPS was treated by oral administration of LQJD. The recruitment of body temperature and the histopathology of lung tissue from all groups were evaluated to grade the severity of the inflammation. The inflammatory cytokine levels, including tumor necrosis factor-α (TNF-α)and interleukin-6 (IL-6), were examined by ELISA assay, and the TLR4 and NF-κBp65 expression levels were evaluated by Real time-PCR and western blotting. It was observed that LQJD reduced the LPS-induced body temperature, inflammatory cytokines level, and lung injuries, and blocked the activation of TLR4/NF-κBp65 signaling in lung tissue. This study demonstrates that LQJD has a protective effect on LPS-induced inflammatory rats through the signaling pathway of TLR4 and NF-κBp65.


Asunto(s)
Lesión Pulmonar Aguda/tratamiento farmacológico , Medicamentos Herbarios Chinos/uso terapéutico , Lipopolisacáridos/toxicidad , FN-kappa B/antagonistas & inhibidores , Transducción de Señal/efectos de los fármacos , Receptor Toll-Like 4/antagonistas & inhibidores , Lesión Pulmonar Aguda/inducido químicamente , Lesión Pulmonar Aguda/metabolismo , Animales , Relación Dosis-Respuesta a Droga , Medicamentos Herbarios Chinos/aislamiento & purificación , Medicamentos Herbarios Chinos/farmacología , Inflamación/inducido químicamente , Inflamación/tratamiento farmacológico , Inflamación/metabolismo , FN-kappa B/metabolismo , Ratas , Ratas Sprague-Dawley , Transducción de Señal/fisiología , Receptor Toll-Like 4/metabolismo
13.
Zhongguo Zhong Yao Za Zhi ; 42(2): 390-395, 2017 Jan.
Artículo en Chino | MEDLINE | ID: mdl-28948749

RESUMEN

The molecular connectivity index was adopted to explore the characteristics of supramolecular imprinting template of herbs distributed to liver meridian, in order to provide scientific basis for traditional Chinese medicines(TCMs) distributed to liver meridian. In this paper, with "12th five-year plan" national planning textbooks Science of Traditional Chinese Medicine and Chemistry of Traditional Chinese Medicine as the blueprint, literatures and TCMSP sub-databases in TCM pharmacology of northwest science and technology university of agriculture and forestry were retrieved to collect and summarize active constituents of TCM distributed to liver meridian, and calculate the molecular connectivity index. The average molecular connectivity index of ingredients distributed to liver meridian was 9.47, which was close to flavonoid glycosides' (9.17±2.11) and terpenes (9.30±3.62). Therefore, it is inferred that template molecule of liver meridian is similar to physicochemical property of flavonoid glycosides and terpenes, which could be best matched with imprinting template of liver meridian.


Asunto(s)
Medicamentos Herbarios Chinos/farmacología , Hígado/fisiología , Medicina Tradicional China , Meridianos , Humanos
14.
Anticancer Res ; 37(8): 4507-4514, 2017 08.
Artículo en Inglés | MEDLINE | ID: mdl-28739746

RESUMEN

BACKGROUND/AIM: Microsatellite instability (MSI) is a hallmark of genomic instability. In gastric cancer (GC), MSI phenotype is an important molecular subgroup that is often closely correlated with elevated mutation rates on the whole genome level. However, on a single gene level, it is still unknown whether the MSI status of a gene is correlated with the mutational profile of the gene itself. MATERIALS AND METHODS: We analyzed intron MSI status and exon mutational profile of TP53 through short tandem repeat (STR) scanning and direct sequencing respectively in gastric cancers and their matched normal tissues. RESULTS: MSI status of the TP53 intron was significantly associated with the mutational profile of seven TP53 exon positions (p=0.0416), male patients (p=0.0095), and drinking (p=0.0474), and showed a mild correlation with longer survival time (p=0.0584) and increasing age (p=0.0611). CONCLUSION: TP53 exons tended to mutation in the status of TP53 intron MSI.


Asunto(s)
Exones , Genes p53 , Intrones , Inestabilidad de Microsatélites , Mutación , Neoplasias Gástricas/genética , Adulto , Anciano , Anciano de 80 o más Años , Análisis Mutacional de ADN , Femenino , Genotipo , Humanos , Masculino , Persona de Mediana Edad , Clasificación del Tumor , Factores de Riesgo , Neoplasias Gástricas/mortalidad , Neoplasias Gástricas/patología
15.
Arch Pharm Res ; 38(2): 282-9, 2015 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-24733674

RESUMEN

Cantharidin (CTD), a chemical compound secreted by blister beetles, has been shown with anti-tumor property in many cancer cells. In this study, our data showed that CTD exerts potent anti-angiogenesis activity in a dose-dependent manner. CTD dose dependently suppressed human umbilical vascular endothelial cells proliferation, migration, and tube formation in vitro. Furthermore, CTD concentration dependently inhibited angiogenesis in chick embryo CAM model in vivo. At the molecular level, CTD abrogated VEGF-induced activation of STAT3 and suppressed the phosphorylation of JAK1 and ERK in a dose-dependent manner. Furthermore, CTD blocked the phosphorylation of AKT in a time-dependent manner. Taken together, these findings clearly demonstrate for the first time that CTD can inhibit angiogenesis and may have applications in the development of new anti-angiogenesis drugs.


Asunto(s)
Inhibidores de la Angiogénesis/farmacología , Cantaridina/farmacología , Janus Quinasa 1/metabolismo , Sistema de Señalización de MAP Quinasas/efectos de los fármacos , Proteínas Proto-Oncogénicas c-akt/metabolismo , Factor de Transcripción STAT3/metabolismo , Factor A de Crecimiento Endotelial Vascular/farmacología , Animales , Técnicas de Cultivo de Célula , Movimiento Celular/efectos de los fármacos , Proliferación Celular/efectos de los fármacos , Embrión de Pollo , Membrana Corioalantoides/irrigación sanguínea , Membrana Corioalantoides/efectos de los fármacos , Relación Dosis-Respuesta a Droga , Células Endoteliales/efectos de los fármacos , Células Endoteliales/metabolismo , Células Endoteliales/patología , Células Endoteliales de la Vena Umbilical Humana , Humanos , Fosforilación , Factor A de Crecimiento Endotelial Vascular/fisiología
16.
Nan Fang Yi Ke Da Xue Xue Bao ; 31(7): 1169-74, 2011 Jun.
Artículo en Chino | MEDLINE | ID: mdl-21764687

RESUMEN

OBJECTIVE: To construct a PCR chip with a gene panel for predicting and diagnosing metastatic colorectal cancer. METHODS: The PCR chip was constructed by integrating 29 genes related to colorectal cancer metastasis identified by gene chip analysis and 3 housekeeping genes into a gene panel. The PCR chip was used for detecting the mRNA expressions of the integrated genes in colorectal cell lines, cancerous specimen and adjacent normal mucosa. The primers for amplification were refined and optimized by several rounds of preliminary reactions. RESULTS: The PCR chip containing the 29 candidate genes and 3 housekeeping genes was successfully constructed, which showed specific amplifications of the genes. The results of the PCR chip for detecting the mRNA of the 29 genes related to colorectal cancer metastasis showed a concordance rate of 86% (25 out of 29) with the gene chip data. Application of the PCR chip in the examination of the clinical specimens identified 15 differentially expressed genes between metastatic colorectal cancer and colorectal cancer without metastasis. CONCLUSION: The constructed PCR chip is reliable in the prediction of metastasis of colorectal cancer, and provides a molecular means for evaluating the prognosis of colorectal cancer metastasis.


Asunto(s)
Neoplasias Colorrectales/genética , Neoplasias Colorrectales/patología , Perfilación de la Expresión Génica , Reacción en Cadena de la Polimerasa/métodos , Regulación Neoplásica de la Expresión Génica , Humanos , Metástasis de la Neoplasia/diagnóstico , Análisis de Secuencia por Matrices de Oligonucleótidos , Pronóstico
17.
Nan Fang Yi Ke Da Xue Xue Bao ; 30(9): 2025-9, 2010 Sep.
Artículo en Chino | MEDLINE | ID: mdl-20855242

RESUMEN

OBJECTIVE: To identify the enhancers of human lung specific X protein (LUNX) and their regulation at the transcription level in vitro. METHODS: Three enhancer fragments (E1:+3770~+3959bp; E2: +6454~+6555bp; E3: +14553~+14652 bp) predicted by bioinformatics software were isolated from the human genomic DNA by PCR amplification. Luciferase assay was performed to detect the activities of the enhancers in transcriptional regulation. RESULTS: PCR products were confirmed by DNA sequencing. The amplified enhancers digested by Kpn I/Xho I and BamH I/Sal I, to generate the sticky-end fragments were inserted into PGL3-promoter in a reporter vector, and 6 luciferase expression vectors were obtained. All the reporter plasmids and pGL3-promoter were transiently transfected into HEK293 cells with an internal control of pSV-ß-Galactosidase reporter vector. The enhancer activity of each construct was evaluated by luciferase assay of the cell extracts after transfection for 48 h. The results showed that the 3 fragments, when located upstream, did not increase transcription of reporter gene, but when at the downstream, E1 and E3 increased the transcription by 2.83 and 1.59 folds of that of pGL3-promoter, respectively. CONCLUSION: LUNX gene sequences from +3770 to +3959 bp and +14553 to +14652 bp possess the capacity to enhance gene transcription.


Asunto(s)
Elementos de Facilitación Genéticos , Glicoproteínas/genética , Fosfoproteínas/genética , Transcripción Genética , Secuencia de Bases , Clonación Molecular , Regulación de la Expresión Génica , Células HEK293 , Humanos , Datos de Secuencia Molecular
18.
Clin Exp Metastasis ; 27(7): 517-27, 2010 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-20617370

RESUMEN

Single cell progenies (SCPs) inherit biological properties from their isogenetic mother cells. If a single cancer cell can give rise to progenies, which can be passaged sustainably in vitro and produce tumor in xenotransplantation, the cell should be cancer initiating cell. CD133 (Prominin-1, Prom1) is the marker of human colorectal cancer (CRC) stem cells and probably a marker of metastatic cancer stem cells (CSCs). Thirty-three SCPs of CRC cell line SW480 were isolated by limited dilution methods, thirty of which are CD133 positive and three negative. All of the CD133(+) SCPs are tumorigenic, and the subcutaneous tumors expanded rapidly, while only 1 of 3 CD133(-) SCPs developed a minimal tumor in nude mice. Orthotopic transplantation experiments showed that CD133(+) SCPs possessed heterogeneity in intestinal wall invasion, lymph node and liver metastases. CD133(+) SCPs varied in cell growth, invasive ability, epithelial-mesenchymal-transition and expression of CSCs markers (CD133, CD44, and CXCR4) associated with metastatic potential. CD133(-) SCPs did not produce secondary transplanted tumor, intestinal invasion and metastasis. The results indicated CD133(+) subpopulation of SW480 SCPs bear heterogeneous invasive and metastatic ability, and CRC-CSCs might be a heterogeous subpopulation.


Asunto(s)
Antígenos CD/inmunología , Neoplasias Colorrectales/patología , Glicoproteínas/inmunología , Invasividad Neoplásica , Metástasis de la Neoplasia , Péptidos/inmunología , Antígeno AC133 , Animales , Secuencia de Bases , Western Blotting , Línea Celular Tumoral , Neoplasias Colorrectales/inmunología , Cartilla de ADN , Citometría de Flujo , Humanos , Inmunohistoquímica , Ratones , Ratones Endogámicos BALB C , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa
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