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1.
Braz Oral Res ; 34: e030, 2020.
Artículo en Inglés | MEDLINE | ID: mdl-32236319

RESUMEN

The abnormal increase in proliferation rate of human periodontal ligament stem cells (PDLSCs) is considered to be involved in the pathogenesis of periodontitis, a disease in which the IL-10-mediated anti-inflammatory pathway plays a critical role. This study aimed to investigate the involvement of microRNA-466l in periodontitis and to explore the possible interaction between IL-10 and microRNA-466l. PDLSCs were obtained from periodontitis-affected teeth and healthy control teeth. The expression of microRNA-466l and IL-10 mRNA was measured in PDLSCs using RT-qPCR. The proliferation ability of PDLSCs was analyzed using CCK-8 assays. Overexpression of microRNA-466l in a PDLSC cell line was established using two different types of PDLSCs, and the effect of microRNA-466l overexpression on IL-10 expression and cell proliferation were detected by western blot and CCK-8 assays, respectively. We found that expression levels of microRNA-466l and IL-10 mRNA were significantly lower (P < 0.05) in PDLSCs derived from periodontitis-affected teeth compared to those derived from healthy teeth. However, the cell proliferation ability was significantly higher in the PDLSCs derived from periodontitis-affected teeth. Meanwhile microRNA-466l overexpression decreased cell proliferation rates of both types of PDLSCs and upregulated IL-10 expression. Together, these data suggest that microRNA-466l can upregulate IL-10 and reduce the proliferation rate of PDLSCs.


Asunto(s)
Proliferación Celular/fisiología , Interleucina-10/metabolismo , Interleucina-10/uso terapéutico , MicroARNs/metabolismo , Periodontitis/terapia , Células Madre/metabolismo , Adulto , Western Blotting , Diferenciación Celular , Humanos , Periodontitis/genética , Periodontitis/metabolismo , Regulación hacia Arriba
2.
Life Sci ; 251: 117595, 2020 Jun 15.
Artículo en Inglés | MEDLINE | ID: mdl-32240681

RESUMEN

AIMS: The activation of hepatic stellate cells (HSCs) plays a central role in liver fibrosis progression. Phospholipase D (PLD) enzymes participate in multiple cellular activities. However, whether and how PLD regulates HSCs activation remain elusive. MAIN METHODS: The expression of intrahepatic PLD1 and PLD2 was determined in CCl4-induced mouse liver fibrosis models by western blot and immunohistochemistry. Cell model of liver fibrogenesis was constructed using rat HSCs line (HSC-T6) treated with recombinant transforming growth factor ß1 (TGFß1). Fibrogenesis was evaluated on the aspects of proliferation, expression of pro-fibrogenic markers and migration. The effects mediated by PLD1-mTOR axis on TGFß1-induced fibrogenesis were evaluated using HSC-T6 treated with small-molecular PLD1 inhibitors, PLD1-SiRNA, rapamycin (mTOR inhibitor) and MHY1485 (mTOR activator). KEY FINDINGS: Significant increase of PLD1, not PLD2 was documented in CCl4-induced cirrhotic compared to normal liver tissues. Suppression of PLD1 activities by PLD inhibitors or down-regulation of PLD1 expression in HSC-T6 could significantly restrain TGFß1-induced fibrogenesis, as reflected by decreased cell proliferation and reduced expression of pro-fibrogenic markers. Besides, either PLD1 inhibitor or PLD1-SiRNA significantly inhibited mTOR activity of HSC-T6. Moreover, PLD1 inhibitors not only exhibited similar effects with rapamycin in TGFß1-induced fibrogenesis, but also blunted MHY1485 enhanced cell proliferation of HSC-T6. SIGNIFICANCE: The PLD1-mTOR axis of HSCs could be therapeutically targeted in advanced liver fibrosis.


Asunto(s)
Células Estrelladas Hepáticas/metabolismo , Cirrosis Hepática/fisiopatología , Fosfolipasa D/metabolismo , Serina-Treonina Quinasas TOR/metabolismo , Animales , Línea Celular , Proliferación Celular/fisiología , Modelos Animales de Enfermedad , Progresión de la Enfermedad , Masculino , Ratones , Ratones Endogámicos C57BL , Ratas
3.
Life Sci ; 249: 117503, 2020 May 15.
Artículo en Inglés | MEDLINE | ID: mdl-32142767

RESUMEN

AIMS: To investigate the role and mechanism of insulin-like growth factor 1(IGF-1)-mediated EMT on multiple myeloma (MM) growth and metastasis. MATERIALS AND METHODS: The expression data from GEO datasets were utilized to explore the expression levels of IGF-1 and epithelial-mesenchymal transition (EMT) markers in MM. Western blotting and flow cytometry analysis were performed to detect the protein levels of EMT markers as well as key components of the PI3K/Akt pathway. Cell proliferation ability was assessed using colony formation assay and EdU incorporation assays. Transwell migration and invasion assays were performed to assess cell metastasis properties. Vimentin was knocked down by using electro-transfection with small interfering RNA (siRNA) to detect the effect of IGF-1-mediated EMT on MM cell growth and metastasis. KEY FINDINGS: First of all, the analysis of GEO database revealed that IGF-1 was excessively expressed and closely correlated with the expression of the EMT markers in MM patients. Furthermore, we demonstrated that IGF-1 enhanced the acquisition of mesenchymal features in a time-dependent manner. Additionally, in vitro studies revealed that IGF-1-mediated mesenchymal phenotype promoted MM migration, invasion and colony formation. Finally, the mechanism study showed PI3K/Akt signaling pathway was involved in the IGF-1-induced EMT in MM cells. SIGNIFICANCE: IGF-1-induced mesenchymal phenotype contributed to MM progression via the PI3K/Akt pathway regulation.


Asunto(s)
Transición Epitelial-Mesenquimal/fisiología , Factor I del Crecimiento Similar a la Insulina/fisiología , Mieloma Múltiple/patología , Fosfatidilinositol 3-Quinasas/metabolismo , Proteínas Proto-Oncogénicas c-akt/metabolismo , Biomarcadores/metabolismo , Línea Celular Tumoral , Proliferación Celular/fisiología , Progresión de la Enfermedad , Regulación hacia Abajo , Humanos , Mieloma Múltiple/metabolismo , Metástasis de la Neoplasia , Transducción de Señal , Regulación hacia Arriba , Vimentina/metabolismo
4.
Anticancer Res ; 40(3): 1535-1542, 2020 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-32132054

RESUMEN

BACKGROUND/AIM: Sessile serrated polyps without dysplasia (SSPND) are characterized by crypts with serrated epithelium, albeit with irregular, corrupted shapes (CCS). PATIENTS AND METHODS: Cell proliferation was explored in the CCS from 60 SSPND and in the crypts from 12 normal colons. Sections were immuno-stained with the Ki-67 proliferation-cell (PC) marker, and with the p53 tumour-suppressor gene. RESULTS: Three predominant PC-phenotypes were found in the CCS from the 60 SSPND: 44 (73.3%) exhibited ectopic, asymmetric, randomly distributed PC-clusters, 12 (20.0%), continuous PC in one or in both slopes of the crypts, and in the remaining 4 (6.7%), single, randomly distributed PC were recorded. In contrast, the scrutiny of more than 200,000 normal colon crypts (controls) showed symmetrically aligned PC, restricted to the lower third of the crypts. p53-up-regulation in CCS was recorded in 11(18.3%) of the 60 NDSSP, but in none of the normal crypts in the 12 controls. CONCLUSION: The non-dysplastic epithelium that lines CCS in SSPND coexists with an asymmetric relocation of the PC-domains. In addition, the CCS in nearly one-fifth of the SSPND exhibited p53-up-regulated cells. Taken together, the non-dysplastic CCS epithelium in SSPND thrives with somatic mutations. The accretion of putative mutated non-dysplastic CCS might be a crucial event in the evolution of colonic SSPND towards sessile serrated adenomas.


Asunto(s)
Pólipos del Colon/patología , Proliferación Celular/fisiología , Forma de la Célula/fisiología , Pólipos del Colon/genética , Pólipos del Colon/metabolismo , Genes p53 , Humanos , Inmunoensayo , Fenotipo , Proteína p53 Supresora de Tumor/biosíntesis , Proteína p53 Supresora de Tumor/genética , Regulación hacia Arriba
5.
Gene ; 740: 144520, 2020 May 25.
Artículo en Inglés | MEDLINE | ID: mdl-32130980

RESUMEN

Osteosarcoma (OS) is the most common primary bone tumor, and mainly occurs in children and early adults. The overall pathogenesis of osteosarcoma remains unclear. Circular RNAs (circRNAs) have been demonstrated to be one of the regulators in tumors as non-coding RNAs. In present study, we identified an elevated expressed circ-0060428 in osteosarcoma cells. The results of CCK-8 and flow cytometry assay showed that circ-0060428 promoted cell proliferation and reduced cell apoptosis of osteosarcoma cells. Bioinformatics analyses predicted the binding site between miR-375 to RPBJ and circ-0060428, and dual luciferase reporter assay verified this prediction. Furthermore, miRNA inhibitor of miR-375 could recover the influence of cell proliferation and apoptosis by knocking down the expression of circ-0060428. Meanwhile, knockdown of both circ-0060428 and RBPJ resulted in a lower apoptosis rate than circ-0060428 alone. Western blot analyses indicated that circ-0060428 regulated the expression of apoptosis related proteins (Bax, Bcl-2, and cleaved-Caspase-3) by upregulating RPBJ expression in osteosarcoma cells. Altogether, we confirmed the up-regulated circ-0060428 could improve the proliferation and survival of osteosarcoma cells by sponging miR-375 to upregulate RBPJ expression. This finding supported a novel clinically biomarker and treatment target for osteosarcoma therapy.


Asunto(s)
Proliferación Celular/fisiología , Proteína de Unión a la Señal Recombinante J de las Inmunoglobulinas/metabolismo , MicroARNs/metabolismo , Osteosarcoma/fisiopatología , ARN Circular , Línea Celular Tumoral , Proliferación Celular/genética , Perfilación de la Expresión Génica , Humanos , Proteína de Unión a la Señal Recombinante J de las Inmunoglobulinas/genética , MicroARNs/genética , Osteosarcoma/metabolismo , ARN Circular/genética , ARN Circular/metabolismo
6.
Anticancer Res ; 40(3): 1307-1314, 2020 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-32132027

RESUMEN

BACKGROUND/AIM: Malignant pleural mesothelioma (MPM) is an intractable cancer, and causes of its malignant transformation are not well known. Adenosine deaminase acting on RNA (ADAR) is an RNA-editing enzyme that converts adenosine into inosine in double-stranded RNAs potentially involved in malignant development. MATERIALS AND METHODS: To examine the role of ADAR1 and ADAR2 in MPM, small interfering RNAs (siRNAs) against ADAR1 or ADAR2 were used. RESULTS: Transfection of siRNA against ADAR2 suppressed proliferation, motility, and invasiveness of MPM cells expressing both ADAR1 and ADAR2; however, siRNA against ADAR1 did not affect these cellular activities. Overexpression of ADAR2, that was incapable of binding to RNA, suppressed growth, motility, and invasion of MPM cells. However, overexpression of ADAR2 that had no enzyme activity did not alter the malignant properties of MPM cells. CONCLUSION: Enhancement of the malignant characteristics of cultured MPM cells via ADAR2 was independent of RNA-editing activity.


Asunto(s)
Adenosina Desaminasa/metabolismo , Neoplasias Pulmonares/genética , Neoplasias Pulmonares/metabolismo , Mesotelioma/genética , Mesotelioma/metabolismo , Edición de ARN , Proteínas de Unión al ARN/metabolismo , Adenosina Desaminasa/biosíntesis , Adenosina Desaminasa/genética , Línea Celular Tumoral , Movimiento Celular/fisiología , Proliferación Celular/fisiología , Humanos , Neoplasias Pulmonares/enzimología , Neoplasias Pulmonares/patología , Mesotelioma/enzimología , Mesotelioma/patología , ARN Interferente Pequeño/administración & dosificación , ARN Interferente Pequeño/genética , Proteínas de Unión al ARN/biosíntesis , Proteínas de Unión al ARN/genética , Transfección
7.
Anticancer Res ; 40(3): 1367-1374, 2020 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-32132033

RESUMEN

BACKGROUND/AIM: Ring finger protein 126 (RNF126) belongs to the family of RING E3 ubiquitin ligases. Although RNF126 has been reported to be overexpressed in several cancers, the role of RNF126 in gastric cancer remains unclear. MATERIALS AND METHODS: We investigated the RNF126 expression in 170 primary gastric cancer tissues by immunohistochemistry, and explored its prognostic impact. The effect of the RNF126 expression on the proliferation of cancer cells was evaluated in vitro. RESULTS: The RNF126 expression was significantly associated with tumor depth and presence of venous invasion. The RNF126 status was identified as an independent prognostic factor (p<0.001). RNF126 gene silencing significantly inhibited the proliferation of gastric cancer cells, induced G1 phase arrest and increased the p21 protein level. CONCLUSION: RNF126 expression has a significant prognostic value in gastric cancer. RNF126 may play an important role in tumor progression of gastric cancer.


Asunto(s)
Neoplasias Gástricas/metabolismo , Ubiquitina-Proteína Ligasas/biosíntesis , Proliferación Celular/fisiología , Humanos , Inmunohistoquímica , Pronóstico , Neoplasias Gástricas/genética , Neoplasias Gástricas/patología , Tasa de Supervivencia , Ubiquitina-Proteína Ligasas/genética
8.
Nat Commun ; 11(1): 1025, 2020 02 24.
Artículo en Inglés | MEDLINE | ID: mdl-32094341

RESUMEN

A bioengineered skeletal muscle construct that mimics structural and functional characteristics of native skeletal muscle is a promising therapeutic option to treat extensive muscle defect injuries. We previously showed that bioprinted human skeletal muscle constructs were able to form multi-layered bundles with aligned myofibers. In this study, we investigate the effects of neural cell integration into the bioprinted skeletal muscle construct to accelerate functional muscle regeneration in vivo. Neural input into this bioprinted skeletal muscle construct shows the improvement of myofiber formation, long-term survival, and neuromuscular junction formation in vitro. More importantly, the bioprinted constructs with neural cell integration facilitate rapid innervation and mature into organized muscle tissue that restores normal muscle weight and function in a rodent model of muscle defect injury. These results suggest that the 3D bioprinted human neural-skeletal muscle constructs can be rapidly integrated with the host neural network, resulting in accelerated muscle function restoration.


Asunto(s)
Bioimpresión/métodos , Regeneración Tisular Dirigida/métodos , Enfermedades Musculares/terapia , Mioblastos Esqueléticos/fisiología , Neuronas/fisiología , Animales , Materiales Biomiméticos/química , Materiales Biomiméticos/uso terapéutico , Diferenciación Celular/fisiología , Proliferación Celular/fisiología , Supervivencia Celular/fisiología , Modelos Animales de Enfermedad , Estudios de Factibilidad , Humanos , Hidrogeles/química , Masculino , Músculo Esquelético/citología , Músculo Esquelético/lesiones , Músculo Esquelético/fisiología , Enfermedades Musculares/fisiopatología , Red Nerviosa/fisiología , Unión Neuromuscular/citología , Unión Neuromuscular/fisiología , Impresión Tridimensional , Ratas , Factores de Tiempo
9.
PLoS One ; 15(2): e0228909, 2020.
Artículo en Inglés | MEDLINE | ID: mdl-32049991

RESUMEN

BACKGROUND/OBJECTIVE: Patients with non-small cell lung cancer (NSCLC) develop resistance to antitumor agents by mechanisms that involve the epithelial-to-mesenchymal transition (EMT). This necessitates the development of new complementary drugs, e.g., cannabinoid receptors (CB1 and CB2) agonists including tetrahydrocannabinol (THC) and cannabidiol (CBD). The combined use of THC and CBD confers greater benefits, as CBD enhances the effects of THC and reduces its psychotropic activity. We assessed the relationship between the expression levels of CB1 and CB2 to the clinical features of a cohort of patients with NSCLC, and the effect of THC and CBD (individually and in combination) on proliferation, EMT and migration in vitro in A549, H460 and H1792 lung cancer cell lines. METHODS: Expression levels of CB1, CB2, EGFR, CDH1, CDH2 and VIM were evaluated by quantitative reverse transcription-polymerase chain reaction. THC and CBD (10-100 µM), individually or in combination (1:1 ratio), were used for in vitro assays. Cell proliferation was determined by BrdU incorporation assay. Morphological changes in the cells were visualized by phase-contrast and fluorescence microscopy. Migration was studied by scratch recolonization induced by 20 ng/ml epidermal growth factor (EGF). RESULTS: The tumor samples were classified according to the level of expression of CB1, CB2, or both. Patients with high expression levels of CB1, CB2, and CB1/CB2 showed increased survival reaching significance for CB1 and CB1/CB2 (p = 0.035 and 0.025, respectively). Both cannabinoid agonists inhibited the proliferation and expression of EGFR in lung cancer cells, and CBD potentiated the effect of THC. THC and CBD alone or in combination restored the epithelial phenotype, as evidenced by increased expression of CDH1 and reduced expression of CDH2 and VIM, as well as by fluorescence analysis of cellular cytoskeleton. Finally, both cannabinoids reduced the in vitro migration of the three lung cancer cells lines used. CONCLUSIONS: The expression levels of CB1 and CB2 have a potential use as markers of survival in patients with NSCLC. THC and CBD inhibited the proliferation and expression of EGFR in the lung cancer cells studied. Finally, the THC/CBD combination restored the epithelial phenotype in vitro.


Asunto(s)
Cannabidiol/metabolismo , Carcinoma de Pulmón de Células no Pequeñas/metabolismo , Proliferación Celular/fisiología , Dronabinol/metabolismo , Transición Epitelial-Mesenquimal/fisiología , Neoplasias Pulmonares/metabolismo , Receptor Cannabinoide CB1/metabolismo , Receptor Cannabinoide CB2/metabolismo , Células A549 , Adulto , Anciano , Anciano de 80 o más Años , Agonistas de Receptores de Cannabinoides/farmacología , Cannabinoides/metabolismo , Línea Celular Tumoral , Proliferación Celular/efectos de los fármacos , Transición Epitelial-Mesenquimal/efectos de los fármacos , Femenino , Expresión Génica/efectos de los fármacos , Humanos , Masculino , Persona de Mediana Edad , Psicotrópicos/farmacología
10.
Anticancer Res ; 40(2): 653-664, 2020 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-32014906

RESUMEN

BACKGROUND/AIM: Peroxisome proliferator-activated receptor γ coactivator 1α (PGC-1α) is a master regulator of mitochondrial biogenesis and metabolism. We investigated the effect of PGC-1α knockdown in the human colorectal cancer cell line SW620, which highly expresses PGC-1α. MATERIALS AND METHODS: We established the PGC-1α shRNA-silenced SW620 stable cell line (PGC-1α shRNA-SW620 cells) and examined cell proliferation by cell counts and carboxyfluorescein succinimidyl ester (CFSE) staining, migration by wound-healing and transwell migration assay, and invasion by transwell assays. RESULTS: PGC-1α knockdown inhibited cell proliferation, migration, and invasion in SW620 cells. Western blot analysis showed that p-AKT, p-GSK-3ß, ß-catenin, N-cadherin and vimentin expression were all reduced, but E-cadherin had increased expression in PGC-1α shRNA-SW620 cells. We also examined cell proliferation, migration, invasion and the expression of p-AKT, p-GSK-3ß, ß-catenin, N-cadherin, vimentin, and E-cadherin in PGC-1α overexpressing SW480 cells (a low PGC-1α expressing line). We observed a complete reversal of the results seen in the knockdown. CONCLUSION: PGC-1α might regulate cell proliferation and invasion via AKT/GSK-3ß/ß-catenin pathway in SW620 and SW480 cells.


Asunto(s)
Glucógeno Sintasa Quinasa 3 beta/metabolismo , Coactivador 1-alfa del Receptor Activado por Proliferadores de Peroxisomas gamma/metabolismo , Proteínas Proto-Oncogénicas c-akt/metabolismo , beta Catenina/metabolismo , Línea Celular Tumoral , Proliferación Celular/fisiología , Neoplasias Colorrectales/metabolismo , Neoplasias Colorrectales/patología , Técnicas de Silenciamiento del Gen , Humanos , Coactivador 1-alfa del Receptor Activado por Proliferadores de Peroxisomas gamma/biosíntesis , Coactivador 1-alfa del Receptor Activado por Proliferadores de Peroxisomas gamma/genética , Transducción de Señal , Transfección
11.
Anticancer Res ; 40(2): 767-777, 2020 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-32014919

RESUMEN

BACKGROUND/AIM: Chicken ovalbumin upstream promoter-transcription factor II (COUP-TFII) plays an important role in cancer. We examined the effect of COUP-TFII overexpression on the proliferation and invasion of the human colorectal cancer SNU-C4 cells. MATERIALS AND METHODS: SNU-C4 cells were stably transfected with COUP-TFII expression plasmid to overexpress COUP-TFII (COUP-TFII-SNU-C4 cells). Cell proliferation, colony-forming ability and transwell invasion assays were performed. To elucidate the underlying molecular mechanism of COUP-TFII action, western blot analysis, p53 shRNA transfection, and Myr-Akt transfection were performed. RESULTS: Cell proliferation and colony-forming ability were significantly inhibited in COUP-TFII-SNU-C4 cells. Western blot analyses demonstrated that while the expression of p53 and PTEN was increased, the p-Akt levels were decreased in COUP-TFII-SNU-C4 cells. Knockdown of p53 partially restored the cell proliferation, but did not reverse the inhibition of invasion. Constitutive activation of Akt via Myr-Akt transfection reversed the inhibited cell proliferation and invasion by COUP-TFII. CONCLUSION: p53 is required for the inhibition of cell proliferation, and decreased phosphorylation of Akt may mediate the inhibition of cell proliferation and invasion by COUP-TFII.


Asunto(s)
Factor de Transcripción COUP II/biosíntesis , Neoplasias Colorrectales/metabolismo , Fosfohidrolasa PTEN/biosíntesis , Proteínas Proto-Oncogénicas c-akt/metabolismo , Proteína p53 Supresora de Tumor/biosíntesis , Factor de Transcripción COUP II/metabolismo , Proliferación Celular/fisiología , Neoplasias Colorrectales/patología , Humanos , Fosfohidrolasa PTEN/metabolismo , Fosforilación , Transfección , Proteína p53 Supresora de Tumor/metabolismo
12.
Invest Ophthalmol Vis Sci ; 61(2): 19, 2020 02 07.
Artículo en Inglés | MEDLINE | ID: mdl-32053728

RESUMEN

Purpose: To analyze the differences in the vitreous cytokine profiles in epiretinal membrane eyes with and without an ectopic inner foveal layer (EIFL). Methods: Sixty eyes with epiretinal membrane (32 eyes without EIFL and 28 eyes with EIFL) were included. The vitreous samples were collected during surgery for epiretinal membrane. The cytokine levels of the vitreous were measured using a multiplex bead analysis. Results: The mean logMAR visual acuity was worse (0.42 vs. 0.37; P = 0.331) and the central foveal thickness was higher in the EIFL group (496.9 µm vs. 434.2 µm; P = 0.007) than they were in the group without EIFL. The mean EIFL thickness was 164.1 ± 67.7 µm in the EIFL group. On multiplex analysis of the vitreous cytokines, the levels of CD163 (21529 pg/dL vs. 10877 pg/dL; P = 0.002) and macrophage colony-stimulating factor (206 pg/dL vs. 159 pg/dL, P = 0.004) were significantly higher in the EIFL group than they were in the group without EIFL. Conclusions: Eyes with EIFL had increased vitreous levels of M2 macrophage markers. The activation of glial cell proliferation by M2 macrophages may contribute to EIFL formation.


Asunto(s)
Citocinas/metabolismo , Membrana Epirretinal/metabolismo , Fóvea Central/química , Macrófagos/metabolismo , Anciano , Anciano de 80 o más Años , Biomarcadores/metabolismo , Proliferación Celular/fisiología , Femenino , Humanos , Masculino , Persona de Mediana Edad , Neuroglía/citología
13.
Life Sci ; 243: 117296, 2020 Feb 15.
Artículo en Inglés | MEDLINE | ID: mdl-31935390

RESUMEN

AIMS: Ovarian cancer (OC) is the most lethal gynecologic malignant tumors all over the world. HOX antisense intergenic RNA myeloid 1 (HOTAIRM1) has been reported as an important regulator in multiple tumors. However, the functions of HOTAIRM1 in OC and its possible molecular mechanisms remain unclear. MAIN METHODS: qRT-PCR analysis was performed to detect the expression levels of HOTAIRM1, miR-106a-5p and ARHGAP24 mRNA in OC tissues and cells. The functional effects of HOTAIRM1, miR-106a-5p and ARHGAP24 on OC cells were determined by MTT, colony formation, flow cytometry and Transwell assays. Luciferase reporter, RIP and RNA pull-down assays were used to examine the interaction between miR-106a-5p and HOTAIRM1 or ARHGAP24. Tumor xenografts were constructed in nude mice to confirm the roles of HOTAIRM1 in OC in vivo. KEY FINDINGS: HOTAIRM1 expression was lowered in OC tumor tissues and cells. Decreased HOTAIRM1 expression was associated with advanced FIGO stages and lymphatic metastasis. Up-regulation of HOTAIRM1 suppressed OC cell proliferation and invasion, and promoted apoptosis. Also, HOTAIRM1 slowed OC tumor growth in vivo. Moreover, HOTAIRM1 could serve as a competing endogenous RNA (ceRNA) of miR-106a-5p to derepress ARHGAP24 expression. HOTAIRM1-mediated inhibitory effect on OC progression was partly reversed following the restoration of miR-106a-5p expression. Furthermore, ARHGAP24 overexpression repressed OC progression in vitro. SIGNIFICANCE: In conclusion, our study showed that HOTAIRM1 suppressed OC progression through derepression of ARHGAP24 by sponging miR-106a-5p. This finding provides novel insights into the mechanisms of HOTAIRM1 in OC and highlights a potential therapeutic strategy for the treatment of OC.


Asunto(s)
Proliferación Celular/fisiología , Proteínas Activadoras de GTPasa/genética , MicroARNs/genética , MicroARNs/fisiología , Invasividad Neoplásica/genética , Neoplasias Ováricas/patología , Animales , Apoptosis/genética , Línea Celular Tumoral , Progresión de la Enfermedad , Femenino , Humanos , Ratones , Ratones Desnudos , Persona de Mediana Edad , Neoplasias Ováricas/genética
14.
Cell Mol Life Sci ; 77(5): 789-805, 2020 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-31897543

RESUMEN

Age-related macular degeneration (AMD) is a complex eye disease underlined by the death of photoreceptors and degeneration of retinal pigment epithelium (RPE) and choriocapillaris (CC). The mechanism(s) responsible for massive and progressive retinal degeneration is not completely known. Senescence, a state of permanent inhibition of cell growth, may be induced by many factors important for AMD pathogenesis and results in senescence-associated secretory phenotype (SASP) that releases growth factors, cytokines, chemokines, proteases and other molecules inducing inflammation and other AMD-related effects. These effects can be induced in the affected cell and neighboring cells, leading to progression of AMD phenotype. Senescent cells also release reactive oxygen species that increase SASP propagation. Many other pathways of senescence-related AMD pathogenesis, including autophagy, the cGAS-STING signaling, degeneration of CC by membrane attack complex, can be considered. A2E, a fluorophore present in lipofuscin, amyloid-beta peptide and humanin, a mitochondria-derived peptide, may link AMD with senescence. Further studies on senescence in AMD pathogenesis to check the possibility of opening a perspective of the use of drugs killing senescent cells (senolytics) and terminating SASP bystander effects (senostatics) might be beneficial for AMD that at present is an incurable disease.


Asunto(s)
Senescencia Celular/fisiología , Coroides/patología , Degeneración Macular/patología , Células Fotorreceptoras/patología , Epitelio Pigmentado de la Retina/patología , Trastornos de la Visión/patología , Proliferación Celular/fisiología , Humanos , Especies Reactivas de Oxígeno/metabolismo
15.
Cell Prolif ; 53(2): e12757, 2020 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-31916327

RESUMEN

OBJECTIVES: To testify that endothelial cells (ECs) induce astrocyte maturation by leukaemia inhibitory factor (LIF) secretion. MATERIALS AND METHODS: In vivo experiments, mice bearing floxed alleles of YAP were crossed with mice expressing a Cre recombinase driven by the endothelial Tek promoter (Tek-Cre) to finally obtain the following three genotypes: YAPf/f , Tek-Cre; YAPf/w , Tek-Cre; and YAPf/f . Retinal vascularization and astrocyte network were evaluated by whole-mount fluorescence and Western blotting. In vitro, experiments were performed in an astrocyte and human microvascular endothelial cell (HMEC-1) coculture model to analyse the mechanisms underlying the effect of endothelial YAP on astrocytes. RESULTS: In vivo, YAPf/f ;Tek-Cre mice showed delayed angiogenesis, sparse vessels and decreased glial fibrillary acidic protein (GFAP)+ astrocytes but aberrant growth of endothelial networks and immature astrocytes (platelet-derived growth factor A, PDGFRA+ astrocytes) overgrowth. In vitro, Yap deletion attenuated the LIF release that delayed the maturation of retinal astrocyte which was consistent with the results of HMEC-1-astrocyte coculture. The effect of YAP overexpression on LIF-LIFR axis in HMEC-1 interferes the GFAP expression of astrocyte. In contrast, LIF protein rescues the astrocytic GFAP expression when EC YAP was inhibited by siRNAs. CONCLUSIONS: We show that EC yes-associated protein (YAP) is not only a critical coactivator of Hippo signalling in retinal vessel development but also plays an essential role in retinal astrocyte maturation by regulating LIF production.


Asunto(s)
Proteínas Adaptadoras Transductoras de Señales/metabolismo , Astrocitos/metabolismo , Factor Inhibidor de Leucemia/metabolismo , Retina/metabolismo , Vasos Retinianos/metabolismo , Factores de Transcripción/metabolismo , Animales , Astrocitos/fisiología , Diferenciación Celular/fisiología , Proliferación Celular/fisiología , Técnicas de Cocultivo/métodos , Células Endoteliales/metabolismo , Células Endoteliales/fisiología , Femenino , Proteína Ácida Fibrilar de la Glía/metabolismo , Humanos , Masculino , Ratones , Ratones Endogámicos C57BL , Neovascularización Patológica/metabolismo , Neovascularización Patológica/patología , Neovascularización Fisiológica/fisiología , Neurogénesis/fisiología , Retina/fisiología , Vasos Retinianos/fisiología
16.
Sci Rep ; 10(1): 1193, 2020 Jan 27.
Artículo en Inglés | MEDLINE | ID: mdl-31988355

RESUMEN

Single-cell dispensing for automated cell isolation of individual cells has gained increased attention in the biopharmaceutical industry, mainly for production of clonal cell lines. Here, machine learning for classification of cell images is applied for 'real-time' cell viability sorting on a single-cell printer. We show that an extremely shallow convolutional neural network (CNN) for classification of low-complexity cell images outperforms more complex architectures. Datasets with hundreds of cell images from four different samples were used for training and validation of the CNNs. The clone recovery, i.e. the fraction of single-cells that grow to clonal colonies, is predicted to increase for all the samples investigated. Finally, a trained CNN was deployed on a c.sight single-cell printer for 'real-time' sorting of a CHO-K1 cells. On a sample with artificially damaged cells the clone recovery could be increased from 27% to 73%, thereby resulting in a significantly faster and more efficient cloning. Depending on the classification threshold, the frequency at which viable cells are dispensed could be increased by up to 65%. This technology for image-based cell sorting is highly versatile and can be expected to enable cell sorting by computer vision with respect to different criteria in the future.


Asunto(s)
Proliferación Celular/fisiología , Supervivencia Celular/fisiología , Células Clonales/metabolismo , Aprendizaje Automático , Redes Neurales de la Computación , Animales , Anticuerpos Monoclonales/uso terapéutico , Células CHO , Simulación por Computador , Cricetulus , Citometría de Imagen , Proteínas Recombinantes/uso terapéutico
17.
Dev Cell ; 52(2): 236-250.e7, 2020 Jan 27.
Artículo en Inglés | MEDLINE | ID: mdl-31991105

RESUMEN

Regulation of embryonic diapause, dormancy that interrupts the tight connection between developmental stage and time, is still poorly understood. Here, we characterize the transcriptional and metabolite profiles of mouse diapause embryos and identify unique gene expression and metabolic signatures with activated lipolysis, glycolysis, and metabolic pathways regulated by AMPK. Lipolysis is increased due to mTORC2 repression, increasing fatty acids to support cell survival. We further show that starvation in pre-implantation ICM-derived mouse ESCs induces a reversible dormant state, transcriptionally mimicking the in vivo diapause stage. During starvation, Lkb1, an upstream kinase of AMPK, represses mTOR, which induces a reversible glycolytic and epigenetically H4K16Ac-negative, diapause-like state. Diapause furthermore activates expression of glutamine transporters SLC38A1/2. We show by genetic and small molecule inhibitors that glutamine transporters are essential for the H4K16Ac-negative, diapause state. These data suggest that mTORC1/2 inhibition, regulated by amino acid levels, is causal for diapause metabolism and epigenetic state.


Asunto(s)
Sistema de Transporte de Aminoácidos A/metabolismo , Blastocisto/metabolismo , Embrión de Mamíferos/citología , Diana Mecanicista del Complejo 2 de la Rapamicina/metabolismo , Proteínas Serina-Treonina Quinasas/metabolismo , Animales , Proliferación Celular/genética , Proliferación Celular/fisiología , Células Madre Embrionarias/citología , Técnicas de Inactivación de Genes , Ratones
18.
DNA Cell Biol ; 39(2): 310-320, 2020 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-31971826

RESUMEN

Renal vascular sclerosis caused by aging plays an important role in the occurrence and development of chronic kidney disease. Clinical studies have confirmed that endurance exercise is able to delay the aging of skeletal muscle and brain tissue. However, to date, few studies have assessed whether endurance exercise is able to improve the occurrence of renal vascular sclerosis caused by natural aging and its related mechanisms. In this study, we investigated the protective effect of aerobic endurance exercise on renal vascular sclerosis in aged mice and its effect on the phosphatidylinositol 3-kinase/protein kinase B/mammalian target of rapamycin (PI3K/AKT/mTOR) pathway. The results suggested that aerobic endurance exercise preserved kidney morphology and renal function. Glomerular basement membrane thickness was evidently increased, podocyte foot processes were effaced in aged mice, and aerobic endurance exercise significantly ameliorated the overall lesion range. The protein expression of vascular endothelial growth factor (VEGF) and JG12 was lower in the senile control group (OC group). The protein expression of VEGF and JG12 was significantly increased after aerobic endurance exercise. Furthermore, aerobic endurance exercise resulted in downregulation of Bax, Caspase 3, IL-6, and senescent cells and upregulation of Bcl-2. The upregulation of PI3K and its downstream signal molecules AKT and mTOR after aerobic endurance exercise was further observed. Our observations indicated that aerobic endurance exercise may inhibit renal vascular sclerosis in aged mice by regulating the PI3K/AKT/mTOR signaling pathway.


Asunto(s)
Fosfatidilinositol 3-Quinasas/metabolismo , Proteínas Proto-Oncogénicas c-akt/metabolismo , Esclerosis/metabolismo , Serina-Treonina Quinasas TOR/metabolismo , Animales , Apoptosis/fisiología , Línea Celular Tumoral , Proliferación Celular/fisiología , Masculino , Ratones Endogámicos C57BL , Condicionamiento Físico Animal/métodos , Circulación Renal/fisiología , Transducción de Señal/fisiología
19.
Braz J Med Biol Res ; 53(1): e8883, 2020.
Artículo en Inglés | MEDLINE | ID: mdl-31939597

RESUMEN

Opa-interacting protein 5 antisense transcript 1 (OIP5-AS1) is one kind of cytoplasmic long non-coding RNA (lncRNA), which has been demonstrated to play a critical function in multiple cancers. However, the detailed mechanism of OIP5-AS1 in the regulation of cervical cancer progression is still obscure. Here, we demonstrated that lncRNA OIP5-AS1 was upregulated in cervical cancer and was correlated with poor prognosis by bioinformatics studies. OIP5-AS1 depletion inhibited cell proliferation and promoted cell apoptosis in cervical cancer cells. Furthermore, we clarified that ROCK1 was the downstream effector of OIP5-AS1 and OIP5-AS1 acted as a molecular sponge of miR-143-3p. Finally, we verified that OIP5-AS1 exerted its function in the regulation of cervical cancer progression via interacting with miR-143-3p to regulate ROCK1 expression. Our study revealed novel mechanisms about how lncRNA OIP5-AS1 executed its function in cervical cancer and thus provided potential therapeutic targets for the disease.


Asunto(s)
Apoptosis/fisiología , Proliferación Celular/fisiología , MicroARNs/metabolismo , ARN Largo no Codificante/metabolismo , Neoplasias del Cuello Uterino/patología , Quinasas Asociadas a rho/metabolismo , Apoptosis/genética , Western Blotting , Línea Celular Tumoral , Proliferación Celular/genética , Femenino , Regulación Neoplásica de la Expresión Génica , Humanos , MicroARNs/genética , ARN Largo no Codificante/genética , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Regulación hacia Arriba , Neoplasias del Cuello Uterino/metabolismo , Quinasas Asociadas a rho/genética
20.
Life Sci ; 243: 117287, 2020 Feb 15.
Artículo en Inglés | MEDLINE | ID: mdl-31926240

RESUMEN

Vascular smooth muscle cell (VSMC) accumulation and endothelial cell dysfunction are associated with pathogenesis of atherosclerosis. Long noncoding RNA taurine up-regulated gene 1 (TUG1) has been reported to play an important role in cardiovascular diseases, including atherosclerosis. However, the regulatory mechanism underlying TUG1 in atherosclerosis is far from understood. VSMC and human umbilical vein endothelial cells (HUVEC) stimulated by oxidized low-density lipoprotein (ox-LDL) were used as cellular model of atherosclerosis. Cell proliferation and apoptosis were detected by CCK-8, flow cytometry and Western blot. The expression levels of TUG1, microRNA (miR)-148b and insulin-like growth factor 2 (IGF2) were measured by quantitative real-time polymerase chain reaction or Western blot. The target association among TUG1, miR-148b and IGF2 was determined by luciferase reporter assay and RNA immunoprecipitation. The expression of TUG1 was increased in ox-LDL-treated VSMC and HUVEC. Silence of TUG1 inhibited proliferation and promoted apoptosis in ox-LDL-treated VSMC but induced proliferation promotion and apoptosis inhibition in HUVEC stimulated by ox-LDL. miR-148b was a target of TUG1 and its knockdown reversed the effect of TUG1 silence on proliferation and apoptosis of VSMC and HUVEC challenged by ox-LDL. IGF2 was a target of miR-148b and miR-148b regulated proliferation and apoptosis in ox-LDL-treated VSMC and HUVEC by targeting IGF2. TUG1 promoted IGF2 protein expression by sponging miR-148b. TUG1 knockdown attenuated ox-LDL-induced injury through regulating proliferation and apoptosis of VSMC and HUVEC by miR-148b/IGF2 axis, providing a novel mechanism for pathogenesis of atherosclerosis.


Asunto(s)
Apoptosis/fisiología , Proliferación Celular/fisiología , Factor II del Crecimiento Similar a la Insulina/fisiología , Lipoproteínas LDL/farmacología , MicroARNs/fisiología , ARN Largo no Codificante/fisiología , Células Cultivadas , Células Endoteliales de la Vena Umbilical Humana , Humanos
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