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1.
Zhongguo Shi Yan Xue Ye Xue Za Zhi ; 29(2): 494-499, 2021 Apr.
Artículo en Chino | MEDLINE | ID: mdl-33812420

RESUMEN

OBJECTIVE: To investigate the effect of long non-coding RNA-TUC338 on the proliferation and migration of lymphoma cells. METHODS: The expression of TUC338 in different lymphoma cells was detected by fluorescence quantitative PCR, cell proliferation by sulforhodamine B (SRB) assay, migration of lymphoma cells by transwell assay, and protein expression in PI3K/AKT signaling pathway by Western blot. RESULTS: The expression levels of TUC338 in lymphoma cells Daudi, U937, BC-3, and Raji significantly increased in comparison with human normal T lymphocytes H9 (t=13.277, 10.103, 16.200, and 26.687, P=0.002, 0.005, 0.001, and 0.000). Compared with NC-siRNA group, the number of cells crossing the chamber of TUC338-siRNA group was significantly reduced (t=30.508, P=0.000), the protein expression levels of p-PI3K and p-AKT significantly decreased (t=16.872 and 18.371, P=0.000 and 0.000), and OD530 absorbance values at 24 h, 48 h, and 72 h were significantly lower (P<0.05). CONCLUSION: The expression of TUC338 significantly increases in lymphoma cells, and silence of TUC338 effectively inhibits the activation of PI3K/AKT signaling pathway, thereby inhibiting the proliferation and migration of lymphoma cells, which has a potential application value in diagnosis and treatment of lymphoma.


Asunto(s)
ARN Largo no Codificante , Línea Celular Tumoral , Movimiento Celular , Proliferación Celular , Humanos , Fosfatidilinositol 3-Quinasas/metabolismo , Proteínas Proto-Oncogénicas c-akt/metabolismo , ARN Largo no Codificante/genética , Transducción de Señal
2.
Nat Commun ; 12(1): 1490, 2021 03 05.
Artículo en Inglés | MEDLINE | ID: mdl-33674568

RESUMEN

The brain of mammals lacks a significant ability to regenerate neurons and is thus particularly vulnerable. To protect the brain from injury and disease, damage control by astrocytes through astrogliosis and scar formation is vital. Here, we show that brain injury in mice triggers an immediate upregulation of the actin-binding protein Drebrin (DBN) in astrocytes, which is essential for scar formation and maintenance of astrocyte reactivity. In turn, DBN loss leads to defective astrocyte scar formation and excessive neurodegeneration following brain injuries. At the cellular level, we show that DBN switches actin homeostasis from ARP2/3-dependent arrays to microtubule-compatible scaffolds, facilitating the formation of RAB8-positive membrane tubules. This injury-specific RAB8 membrane compartment serves as hub for the trafficking of surface proteins involved in astrogliosis and adhesion mediators, such as ß1-integrin. Our work shows that DBN-mediated membrane trafficking in astrocytes is an important neuroprotective mechanism following traumatic brain injury in mice.


Asunto(s)
Astrocitos/metabolismo , Lesiones Traumáticas del Encéfalo/metabolismo , Cicatriz/metabolismo , Neuropéptidos/genética , Neuropéptidos/metabolismo , Complejo 2-3 Proteico Relacionado con la Actina , Actinas/metabolismo , Animales , Encéfalo/metabolismo , Encéfalo/patología , Lesiones Encefálicas/metabolismo , Lesiones Encefálicas/patología , Lesiones Traumáticas del Encéfalo/patología , Movimiento Celular , Sistema Nervioso Central/metabolismo , Modelos Animales de Enfermedad , Gliosis/metabolismo , Gliosis/patología , Ratones , Ratones Noqueados , Neuroprotección , Transcriptoma , Proteínas de Unión al GTP rab/metabolismo
3.
Zhonghua Yi Xue Za Zhi ; 101(13): 956-965, 2021 Apr 06.
Artículo en Chino | MEDLINE | ID: mdl-33789378

RESUMEN

Objective: To investigate the role of microRNA-139-5p (miR-139-5p) in the occurrence and development of esophageal squamous cell carcinoma (ESCC) and its effects on cell proliferation and invasion of ESCC cells and its molecular mechanisms. Methods: Seventy-five cases of ESCC tissues and paired normal tissues were obtained from thoracic surgery of the First Affiliated Hospital of Zhengzhou University from February 2017 to March 2018. Experiment was divided into two group: ESCC (n=75) and normal esophageal tissues (n=75).GEO datasets and real-time quantitative PCR (qRT-PCR) were used to detect the expression of miR-139-5p in ESCC tissues and cells. miR-139-5p inhibitor, miR-139-5p mimic, negative control, control siRNA, T-box transcliption factor 1(TBX1) siRNA, pcDNA3.1 and pcDNA3.1-TBX1 were transfected into ESCC Eca109 and TE1 cells. qRT-PCR was used to detect the expressions of miR-139-5p and TBX1 in transfected ESCC cells. Cell counting kit 8 (CCK-8) and Transwell chamber were employed to detect cell proliferation and invasion of ESCC cells, respectively. Dual-Luciferase Reporter assay was used to analyze the interaction between miR-139-5p with TBX1. qRT-PCR, Western blot and immunohistochemistry were utilized to detect the expression of TBX1 in ESCC tissues. Western blot was used to detect the expressions of E-cadherin, N-cadherin and Vimentin after transfection. Results: The level of miR-139-5p in ESCC tissues was significantly lower than that in normal tissues (1.17±0.43 vs 5.16±3.62,P<0.001). Log-rank test showed that the survival rate of ESCC patients with high miR-139-5p level (n = 43) was significantly higher than that with low miR-139-5p level (n=32) (67.44% vs 25.00%, P = 0.005). The expression level of miR-139-5p in ESCC cells was significantly lower than that of normal esophageal epithelial cell Het-1A (all P<0.001). The proliferation and invasion ability of ECA109 and TE1 cells with high expression of miR-139-5p were significantly lower than those transfected with negative control (NC) (all P<0.05). Dual-Luciferase Reporter assay showed that miR-139-5p could bind to the 3'-untranslated region of TBX1. miR-139-5p mimic or inhibitor suppressed or promoted the expression of TBX1 protein in Eca109 and TE1 cells, respectively (all P<0.05). Downregulation of TBX1 significantly suppressed proliferation and invasion of ECA109 and TE1 cells, while overexpression of TBX1 significantly promoted proliferation and invasion of ECA109 and TE1 cells (all P<0.05). In addition, pcDNA3.1-TBX1 partially reversed the inhibition of miR-139-5p-mediated invasion ability (all P<0.05), while TBX1 siRNA partially reversed the enhancement of miR-139-5p inhibitor-mediated invasion ability (all P<0.05). Conclusion: miR-139-5p suppressed ESCC cell proliferation and invasion by targeting TBX1.


Asunto(s)
Neoplasias Esofágicas , Carcinoma de Células Escamosas de Esófago , Neoplasias de Cabeza y Cuello , MicroARNs , Línea Celular Tumoral , Movimiento Celular , Proliferación Celular , Neoplasias Esofágicas/genética , Carcinoma de Células Escamosas de Esófago/genética , Regulación Neoplásica de la Expresión Génica , Humanos , MicroARNs/genética , Invasividad Neoplásica
4.
Int J Nanomedicine ; 16: 1961-1976, 2021.
Artículo en Inglés | MEDLINE | ID: mdl-33727809

RESUMEN

Introduction: Metastatic breast cancer seriously harms women's health and is currently the tumour type with the highest mortality rate in women. Recently, the combinatorial therapeutic approaches that integrate anti-cancer drugs and genetic agents is an attractive and promising strategy for the treatment of metastatic breast cancer. Moreover, such a combination strategy requires better drug carriers that can effectively deliver the cargo to the breast cancer cells and achieve controlled release in the cells to achieve better therapeutic effects. Methods: The tumour-targeted and redox-responsive mesoporous silica nanoparticles (MSNs) functionalised with DNA aptamers (AS1411) as a co-delivery system was developed and investigated for the potential against metastatic breast cancer. Doxorubicin (Dox) was loaded onto the MSNs, while AS1411 and a small interfering RNA (siTIE2) were employed as gatekeepers via attachment to the MSNs with redox-sensitive disulfide bonds. Results: The controlled release of Dox and siTIE2 was associated with intracellular glutathione. AS1411 mediated the targeted delivery of Dox by increasing its cellular uptake in metastatic breast cancer, ultimately resulting in a lower IC50 in MDA-MB-231 cells (human breast cancer cell line with high metastatic potency), improved biodistribution in tumour-bearing mice, and enhanced in vivo anti-tumour effects. The in vitro cell migration/invasion assay and in vivo anti-metastatic study revealed synergism in the co-delivery system that suppresses cancer cell metastasis. Conclusion: The tumour-targeted and redox-responsive MSN prepared in this study are promising for the effective delivery and controlled release of Dox and siTIE2 for improved treatment of metastatic breast cancer.


Asunto(s)
Neoplasias de la Mama/tratamiento farmacológico , Doxorrubicina/uso terapéutico , Nanopartículas/química , ARN Interferente Pequeño/administración & dosificación , Dióxido de Silicio/química , Animales , Antineoplásicos/farmacología , Antineoplásicos/uso terapéutico , Neoplasias de la Mama/patología , Muerte Celular/efectos de los fármacos , Línea Celular Tumoral , Movimiento Celular/efectos de los fármacos , Supervivencia Celular/efectos de los fármacos , Preparaciones de Acción Retardada/farmacología , Preparaciones de Acción Retardada/uso terapéutico , Doxorrubicina/farmacología , Portadores de Fármacos/química , Endocitosis/efectos de los fármacos , Femenino , Células HEK293 , Humanos , Ratones Endogámicos BALB C , Ratones Desnudos , Nanopartículas/ultraestructura , Invasividad Neoplásica , Metástasis de la Neoplasia , Oxidación-Reducción , Porosidad , ARN Interferente Pequeño/farmacología , Distribución Tisular/efectos de los fármacos
5.
Int Heart J ; 62(2): 371-380, 2021 Mar 30.
Artículo en Inglés | MEDLINE | ID: mdl-33731513

RESUMEN

Coronary artery disease (CAD) is one of the heavy health burdens worldwide. Aberrant proliferation of vascular smooth muscle cells (VSMCs) contributes to the occurrence and development of CAD. This study aimed at exploring differentially expressed microRNAs (miRNAs) and their regulatory mechanisms in the development of CAD.The miRNA expression profile of GSE28858 was obtained from the Gene Expression Omnibus database. Differentially expressed miRNAs (DEmiRNAs) between CAD and healthy control samples were analyzed using limma package in R. Target genes of DEmiRNAs were predicted, and a miRNA-target gene network was constructed. The relationship between miR-665 and transforming growth factor beta receptor 1 (TGFBR1) was selected for further analysis. The interaction between miR-665 and TGFBR1 was confirmed by dual luciferase reporter assay. Effects of miR-665 on cell viability and apoptosis of VSMCs were evaluated by cell counting kit-8 (CCK-8) assay and flow cytometry, respectively. Besides, western blot assays for BCL2L11 and caspase 3 were also conducted.A total of 38 upregulated miRNAs and 28 downregulated miRNAs were identified. The expression level of miR-665 was significantly downregulated in patients with CAD. TGFBR1 was proved to be a target gene of miR-665. Besides, ectopic expression of miR-665 obviously inhibited VSMC growth and promoted VSMC apoptosis. TGFBR1 overexpression in VSMCs transfected with miR-665 mimic could restore the effect of miR-665 on the proliferation and apoptosis of VSMCs.MiR-665 might participate in the proliferation and apoptosis of VSMCs by targeting TGFBR1.


Asunto(s)
Enfermedad de la Arteria Coronaria/genética , Regulación de la Expresión Génica , MicroARNs/genética , Miocitos del Músculo Liso/metabolismo , Receptor Tipo I de Factor de Crecimiento Transformador beta/genética , Apoptosis , Movimiento Celular , Proliferación Celular , Células Cultivadas , Enfermedad de la Arteria Coronaria/metabolismo , Enfermedad de la Arteria Coronaria/patología , Regulación hacia Abajo , Femenino , Humanos , Masculino , MicroARNs/biosíntesis , Miocitos del Músculo Liso/patología , Receptor Tipo I de Factor de Crecimiento Transformador beta/biosíntesis , Regulación hacia Arriba
6.
Int Heart J ; 62(2): 396-406, 2021 Mar 30.
Artículo en Inglés | MEDLINE | ID: mdl-33731537

RESUMEN

Endothelial injury and inflammation have been found to be essential in the pathogenesis of coronary artery disease (CAD). Circulating exosomes are of great value as novel biomarkers for CAD. However, the role of circulating exosomes in the pathogenesis of CAD remains unclear. Thus, in this study, we aimed to examine whether circulating exosomes from CAD are involved in the endothelial injury and inflammation. The serum-derived exosomes were isolated from CAD and controls using an ExoQuick reagent, and these were then quantified by measuring the protein levels using BCA methods. The uptake of exosomes by human umbilical vein endothelial cells (HUVECs) was observed by laser scanning microscope and analyzed via flow cytometry. Then, HUVECs were treated with vehicle, exosomes from CAD (CAD-exo), and controls (ctrl-exo) in the absence and presence of vascular endothelial growth factor (VEGF). Cell viability, migration, and angiogenesis were evaluated using CCK-8 assay, scratch assay, and tube formation assay. Inflammatory factors including IL-1ß, IL-6, TNF-α, ICAM-1, and VCAM-1 levels were detected via qPCR. As per our findings, no significant differences were noted in uptake of ctrl-exo and CAD-exo by HUVECs. CAD-exo suppressed cell viability in a dose-dependent manner. Compared with ctrl-exo, CAD-exo-treated HUVECs significantly suppressed migration and angiogenesis. However, CAD-exo had a stronger inhibitory effect on VEGF-induced migration and angiogenesis compared with ctrl-exo. Moreover, IL-1ß, TNF-α, and ICAM-1 were determined to be significantly upregulated in HUVECs treated with CAD-exo, but IL-6 and VCAM-1 expressions were not affected. Overall, our results suggest that CAD-exo are involved in endothelial injury and inflammation, which may, in turn, cause endothelial dysfunction and potentially promote the development of CAD.


Asunto(s)
Enfermedad de la Arteria Coronaria/sangre , Exosomas/metabolismo , Movimiento Celular , Proliferación Celular , Células Cultivadas , Enfermedad de la Arteria Coronaria/patología , Femenino , Citometría de Flujo , Células Endoteliales de la Vena Umbilical Humana/metabolismo , Células Endoteliales de la Vena Umbilical Humana/patología , Humanos , Inflamación/metabolismo , Inflamación/patología , Masculino , Persona de Mediana Edad
7.
Biomed Environ Sci ; 34(3): 213-221, 2021 Mar 20.
Artículo en Inglés | MEDLINE | ID: mdl-33766217

RESUMEN

Objective: Cervical cancer (CC) is one of the most common malignant tumors in gynecology. This study aimed to investigate the prognostic significance of serum microRNA (miR)-378a-3p in CC and the effect of miR-378a-3p on tumor growth. Methods: Real-time quantitative polymerase chain reaction analysis was used to measure the expression of miR-378a-3p in serum from patients with CC and healthy control subjects as well as from CC tissues and adjacent normal tissues. The association between serum miR-378a-3p levels and clinicopathological factors was analyzed. The correlation between miR-378a-3p levels and overall survival (OS) of CC patients was determined by Kaplan-Meier analysis. The CC cell proliferation and migration abilities after transfection of miR-378a-3p mimics were detected by Cell Counting Kit-8 and scratch wound healing assays, respectively. Tumor volume and weight in mice treated with miR-378a-3p were measured using a caliper and an electronic balance. Results: MiR-378a-3p expression was downregulated in the serum and tissues of CC patients compared to that in healthy control subjects and normal tissues, respectively. Low expression of miR-378a-3p was positively correlated with large tumor size, advanced tumor stage, and lymph node metastasis. The OS of patients with low expression of miR-378a-3p was significantly lower than that of patients with high expression. Overexpression of miR-378a-3p suppressed the proliferation and migration of CC cells. In vivostudies indicated that overexpression of miR-378a-3p was associated with decreased tumor volume and weight in mice. Conclusion: MiR-378a-3p downregulation is associated with the development and prognosis of CC, suggesting that it may be a potential biomarker for CC.


Asunto(s)
Regulación hacia Abajo , Regulación Neoplásica de la Expresión Génica , MicroARNs/sangre , Neoplasias del Cuello Uterino/metabolismo , Animales , Biomarcadores/sangre , Movimiento Celular , Proliferación Celular , Femenino , Humanos , Ratones , Ratones Endogámicos BALB C , Persona de Mediana Edad
8.
Zhonghua Zhong Liu Za Zhi ; 43(3): 299-305, 2021 Mar 23.
Artículo en Chino | MEDLINE | ID: mdl-33752309

RESUMEN

Objective: To explore the role and molecular mechanism of trophoblastic cell surface antigen 2 (Trop2) in the invasion and migration of ovarian cancer. Methods: Through the data mining of Cancer Cell Line Encyclopedia and TCGA database, the clinical significance of Trop2 expression was analyzed. Western blot was used to detect Trop2 protein expression in ovarian cancer cell lines including A3O, A1780 and SKOV3. SKOV3 cells were used to construct Trop2-short hairpin RNA (shRNA) cell model. Quantitative reverse transcriptase-polymerase chain reaction (qRT-PCR) was used to detect the SKOV3 mRNA expression in SKOV3-shRNA and SKOV3-NC cells. Cell counting kit-8 (CCK8) was used to detect the proliferation of SKOV3-shRNA cells and SKOV3-NC cells. Flow cytometry was used to detect cell cycle and apoptosis in two groups of cells. Transwell array was used to detecte the invasion and migration of SKOV3-shRNA cells and SKOV3-NC cells. Western blot was used to detect the protein expressions of AKT, p-AKT, ß-catenin, caspase3, bcl-2, E-cadherin and vimentin. Results: Trop2 mRNA highly expressed in ovarian cancer, and was related to the tumor stage and patient survival. Compared with A3O cells, Trop2 overexpressed in A1780 and SKOV3 cells (P<0.05). The relative expression levels of Trop2 mRNA in SKOV3-NC group and SKOV3-shRNA group were 1.18±0.24 and 0.42±0.08, with statistically significant difference (P<0.05). The results of CCK-8 array showed that the cell viability of SKOV3-NC group was significantly higher than that of SKOV3-shRNA group (P<0.05). The proportion of G(0)/G(1) cells in SKOV3-NC and SKOV3-shRNA groups were (38.67±4.22)% and (60.24±8.17)%, respectively. G(0)/G(1) arrest was observed in SKOV3-shRNA cells (P<0.05). The apoptosis rate of SKOV3-shRNA group was (26.32±1.81)%, significantly higher than (6.54±1.32)% of SKOV3-NC group (P<0.05). The number of migrating SKOV3 cells in the SKOV3-shRNA and SkOV3-NC groups were 1 255.83±108.44 and 1 679.71±213.92, while the number of invading cells were 242.49±52.09 and 473.54±73.11, respectively. Compared with the SKOV3-NC group, the number of migrating and invading SKOV3-shRNA group was significantly reduced (all P<0.05). The expressions of p-AKT2, Bcl-2, vimentin and ß-catenin were down-regulated, and the expressions of caspase 3 and E-cadherin were up-regulated in SKOV3-shRNA cells. There was no significant change in the total protein level of AKT. Conclusions: Trop2 expression is related to ovarian cancer stage and postoperative survival. Trop2 can promote ovarian cancer cell proliferation and metastasis by activating the AKT/ß-catenin signaling pathway and knockdown of Trop2 inhibits the progression of ovarian cancer.


Asunto(s)
Neoplasias Ováricas , Antígenos de Superficie , Apoptosis , Carcinoma Epitelial de Ovario , Línea Celular Tumoral , Movimiento Celular , Proliferación Celular , Femenino , Humanos , Neoplasias Ováricas/genética
9.
Hua Xi Kou Qiang Yi Xue Za Zhi ; 39(1): 81-87, 2021 Feb 01.
Artículo en Inglés, Chino | MEDLINE | ID: mdl-33723941

RESUMEN

OBJECTIVES: A study was conducted to investigate the molecular mechanism of chromodomain helicase/ATPase DNA binding protein 1-like gene (CHD1L) influencing the invasion and metastasis of tongue squamous cell carcinoma and to provide a new target for clinical inhibition of invasion and metastasis of tongue squamous cell carcinoma. METHODS: Ualcan website was used to analyze the expression of CHD1L in normal epithelial tissue and primary head and neck squamous cell carcinoma and to analyze the effect of lymph node metastasis on the expression of CHD1L in tissues with head and neck squamous cell carcinoma. The relationship between CHD1L expression and the survival rate of patients with head and neck squamous cell carcinoma was tested by the GEPIA website. Western blot was used to quantify the levels of CHD1L protein in human tongue squamous cell carcinoma CAL27 and immortalized human skin keratinocyte cell HaCaT. After knocking down CAL27 in human tongue squamous cell carcinoma cells with an RNA interference plasmid, the cells were designated as SiCHD1L/CAL27 and Scr/CAL27. Western blot was utilized to detect the expression of CHD1L in each group of cells. The change in CAL27 cell proliferation ability was tested by EdU proliferation test after CHD1L knockdown. The change of cell migration ability of each group cells was tested through the wound healing assay. Western blot was used to detect epithelial-mesenchymal transition (EMT) marker E-cadherin and Vimentin protein expression levels. RESULTS: Ualcan database showed that the expression of CHD1L in primary head and neck squamous cell carcinoma tissues was higher than in normal epithelial tissues and in head and neck squamous cell carcinoma tissues with lymph node metastasis. GEPIA website analysis showed that the overall survival rate of patients with head and neck squamous cell carcinoma with high expression of CHD1L was significantly lower than that of patients with low expression. Western blot results showed that CHD1L expression in human tongue squamous carcinoma cells CAL27 was higher than that of human normal skin cells HaCaT. CHD1L expression in SiCHD1L/CAL27 cells was much lower than that in Scr/CAL27 cells. Results of EdU proliferation experiments showed the significant reduction in the cell proliferation ability of the SiCHD1L/CAL27 cells. Results of the wound healing experiments showed the reduction in the migration capacity of the SiCHD1L/CAL27 cells. The expression of E-cadherin increased, whereas that of Vimentin decreased, in SiCHD1L/CAL27 cells. CONCLUSIONS: CHD1L promoted the EMT, proliferation, migration, and invasion ability of tongue squamous cell carcinoma cells.


Asunto(s)
Carcinoma de Células Escamosas , Neoplasias de Cabeza y Cuello , Neoplasias de la Lengua , Adenosina Trifosfatasas , Carcinoma de Células Escamosas/genética , Línea Celular Tumoral , Movimiento Celular , Proliferación Celular , ADN Helicasas , Proteínas de Unión al ADN , Transición Epitelial-Mesenquimal , Regulación Neoplásica de la Expresión Génica , Humanos , Invasividad Neoplásica/genética , Lengua , Neoplasias de la Lengua/genética
10.
J Biomed Nanotechnol ; 17(1): 100-114, 2021 Jan 01.
Artículo en Inglés | MEDLINE | ID: mdl-33653500

RESUMEN

Ionizing radiation (IR) therapy for malignant tumors can damage adjacent tissues, leading to severe wound complications. Plasma-derived exosome treatment has recently emerged as a safe and impactful cell-free therapy. Herein, we aimed to determine whether plasma-derived exosomes could improve the healing of post-radiation wound. Rat plasma-derived exosomes (RP-Exos) were locally injected on cutaneous wounds created on the backs of irradiated rats and boosted the healing process as well as the deposition and remodeling of the extracellular matrix with collagen formation. Subsequently, the effects of RP-Exos were further evaluated on irradiated fibroblasts in vitro. The results suggested that exosomes promoted fibroblast proliferation, migration, cell cycle progression, and cell survival. Moreover, transcriptome sequencing analysis and quantitative polymerase chain reaction validation were performed to identify potential mechanisms. RPExos enhanced the expression of cell proliferation and radioresistance-related genes, and yet downregulated ferroptosis pathway in irradiated fibroblasts. Inhibition of ferroptosis by RP-Exos was further confirmed through colorimetric assay, fluorescence probe and flow cytometry in ferroptosis-induced fibroblasts. Our results suggest that RP-Exos regulate cell proliferation and ferroptosis in irradiated fibroblasts, thereby boosting the healing of irradiated wound. These findings support plasma-derived exosomes as a potential therapeutic method for post-radiation wound complications.


Asunto(s)
Exosomas , Ferroptosis , Células Madre Mesenquimatosas , Animales , Movimiento Celular , Proliferación Celular , Fibroblastos , Plasma , Ratas
11.
Artículo en Chino | MEDLINE | ID: mdl-33691362

RESUMEN

Objective: To investigate the inhibitory effect and molecular mechanism of microRNA-30d (miR-30d) in the process of proliferation, migration and invasion of malignant mesothelioma cell line MSTO-211H. Methods: In April 2017, the human MSTO-211H cells was used to establish miR-30d overexpressed MSTO-211H cell model by transfection of miR-30d mimics. The qRT-PCR was performed to detect the expression level of miR-30d in the cells transfected miR-30d mimics. The effects of miR-30d on the proliferation, apoptosis, migration and invasion of MSTO-211H cells were analyzed by CCK-8 experiment, flow cytometry, cell scratch experiment and Transwell method. Results: After transfection of miR-30d, the expression level of miR-30d in the MSTO-211H+miR-30d cells group was significantly higher than MSTO-211H+miR NC cells group (P<0.01) . The cell activity of MSTO-211H+miR-30d group (105.13%±2.35%) was significantly lower than MSTO-211H+miR NC cells group (115.40%±1.35%) , and the level of apoptosis (3.97%±0.36%) was significantly higher than MSTO-211H+miR NC cells group (1.47%±0.10%) (P<0.01) . The relative migration areas at 12 and 24 h of MSTO-211H+miR-30d cells group (9.35±3.16 µm(2) and 58.19±1.82 µm(2)) were significantly lower than MSTO-211H+miR NC cells group (54.42±5.26 µm(2) and 88.32±1.96 µm(2)) (P<0.01) . Compared with the MSTO-211H+miR NC cells group, the numbers of cell migration and cell invasion were reduced in the MSTO-211H+miR-30d cells group (P<0.01) . Conclusion: miR-30d can regulate the progression of malignant pleural mesothelioma by inhibiting the proliferation, apoptosis, migration and invasion of MSTO-211H cells.


Asunto(s)
Regulación Neoplásica de la Expresión Génica , MicroARNs , Apoptosis , Línea Celular Tumoral , Movimiento Celular , Proliferación Celular , Humanos , MicroARNs/genética , Invasividad Neoplásica
12.
Zhong Nan Da Xue Xue Bao Yi Xue Ban ; 46(1): 18-24, 2021 Jan 28.
Artículo en Inglés, Chino | MEDLINE | ID: mdl-33678632

RESUMEN

OBJECTIVES: To investigate the effects of propofol on the proliferation and invasion of glioma U87 cells and to explore the possible anti-tumor mechanisms. METHODS: The glioma U87 cells was divided into a blank group, a positive control group, and the propofol groups (1.00, 2.00 or 5.00 mmol/L). Cell counting kit-8 (CCK-8) was used to detect cell proliferation; Transwell method was used to detect the effect of propofol on invasion and migration of U87 cells; real-time PCR was used to detect the expression of microRNA-134 (miR-134); Western blotting was used to detect the expression levels of reproduction-related protein Ki-67, invasion-related protein metalloproteinase-2 (MMP-2), metalloproteinase-9 (MMP-9) and phosphatidylinositol 3 kinase (PI3K)/protein kinase B (Akt) signaling pathway-related protein. RESULTS: Compared with the blank group, the proliferation, invasion and migration capacity of U87 cells were reduced in the positive control group and the propofol groups after 48 hours (all P<0.05), along with the decreased expression of Ki-67, MMP-2 and MMP-9 and the ratio of p-PI3K/PI3K and p-Akt/Akt (all P<0.05), while the level of miR-134 was increased significantly (P<0.05). Compared with the positive control group and the 1.00 mmol/L propofol-treated group, the proliferation, invasion and migration capacity of U87 cells, the expression of Ki-67, MMP-2 and MMP-9, and the ratio of p-PI3K/PI3K and p-Akt/Akt was decreased significantly after 48 hours (all P<0.05), while the level of miR-134 was increased significantly in the 2.00 and 5.00 mmol/L propofol-treated groups (both P<0.05). Compared with the 2.00 mmol/L propofol-treated group, the proliferation, invasion and migration capacity of U87 cells, the expression of Ki-67, MMP-2 and MMP-9, and the ratio of p-PI3K/PI3K and p-Akt/Akt was decreased significantly after 48 hours in the 5.00 mmol/L propofol-treated group (all P<0.05), while the level of miR-134 was increased significantly (P<0.05). CONCLUSIONS: Propofol can decrease the proliferation rate, and the invasion and migration abilities of U87 cells, which may be achieved by up-regulation of miR-134 and suppression of PI3K/Akt signaling pathway.


Asunto(s)
Glioma , MicroARNs , Propofol , Línea Celular Tumoral , Movimiento Celular , Proliferación Celular , Glioma/genética , Humanos , Metaloproteinasa 2 de la Matriz/genética , MicroARNs/genética , Fosfatidilinositol 3-Quinasas/genética , Propofol/farmacología , Proteínas Proto-Oncogénicas c-akt/genética
13.
Zhonghua Nei Ke Za Zhi ; 60(4): 362-367, 2021 Apr 01.
Artículo en Chino | MEDLINE | ID: mdl-33765707

RESUMEN

Objective: To investigate the effect of focal adhesion kinase related non kinase (FRNK) on the activation and migration of hepatic stellate cells (HSCs). Methods: Human liver tissue was divided into healthy control group and fibrosis group from March 2019 to September 2019 in Affiliated Hospital of Guizhou Medical University. C57BL/6 mice were divided into wild type (WT) and FRNK gene knockout type (FRNK-/-) groups. The liver fibrosis model was established with carbon tetrachloride (CCl4). After that, FRNK gene overexpression (Ad-FRNK) was constructed with adenovirus vector. HE and Masson staining were used to evaluate the pathological changes and fiber deposition of liver tissue. Western blot was used to detect the expression of PY397-FAK and α-SMA protein. Mouse primary HSCs were extracted, and the effect of FRNK on HSCs migration was detected by wound healing, activation of Rac and Rho was detected by Western blot. Results: The expression of PY397-FAK protein in human liver tissue with hepatic fibrosis was significantly higher than that in healthy control group (0.88±0.09 vs. 0.73±0.09). FRNK was significantly lower than that in control group(0.68±0.09 vs. 0.79±0.11). After animal model was set up, the degree of liver fibrosis in FRNK-/-mice (153±13)% was more serious than that in WT (100%) group. The expression of PY397-FAK and α-SMA protein was significantly elevated (2.50±0.23 vs. 0.75±0.09, 1.46±0.20 vs. 0.92±0.10). After FRNK gene was re-expressed (100%), the degree of liver fibrosis was mainly reversed [(74±6)%], and the expression of PY397-FAK and α-SMA was accordingly decreased(0.68±0.11 vs. 1.12±0.19,0.68±0.10 vs. 0.85±0.06). In vitro, FRNK inhibited the migration of HSCs [WT∶FRNK-/-∶Ad-FRNK,(339±49)%∶(580±53)%∶(259±33)%] and the activation of Rac and Rho proteins (Rac: 0.54±0.07 vs. 0.91±0.10 vs. 0.77±0.12,Rho:0.45±0.05 vs. 0.64±0.06 vs. 0.53±0.07), all P<0.01. Conclusions: FRNK can inhibit the activation and migration of HSCs which contributed to liver fibrosis. The potential mechanism is related to down regulation of PY397-FAK and inhibition of Rac and Rho activation.


Asunto(s)
Células Estrelladas Hepáticas , Cirrosis Hepática , Animales , Movimiento Celular , Regulación hacia Abajo , Proteína-Tirosina Quinasas de Adhesión Focal/metabolismo , Células Estrelladas Hepáticas/metabolismo , Ratones , Ratones Endogámicos C57BL
14.
Zhonghua Shao Shang Za Zhi ; 37(3): 296-300, 2021 Mar 20.
Artículo en Chino | MEDLINE | ID: mdl-33765727

RESUMEN

Wound healing is a complex and critical process, which includes three stages: inflammation, proliferation, and remodeling. The epidermal cells are precisely regulated in this process. On one hand, keratinocytes around the wound edge migrate and proliferate to form a new basement membrane to cover the wound. On the other hand, the epidermal stem cells are activated with the proliferation and differentiation being enhanced, and the terminal differentiation and apoptosis being inhibited; and together with keratinocytes, epidermal stem cells promote the process of re-epithelialization under the regulation of various factors. In the epidermis, there is a group of resident T cell subsets, dendritic epidermal lymphocytes (DETCs) that play a key role in protecting the function of epidermal tissue. DETCs are activated after recognizing unknown antigens, the activated DETCs secret cytokines such as insulin-like growth factor Ⅰ, keratinocyte growth factor-1/2, granulocyte-macrophage colony stimulating factor, interferon-γ, and transforming growth factor-ß, which promote epidermal homeostasis and re-epithelialization by regulating the dynamic balance among keratinocytes migration, proliferation, and apoptosis, and the differentiation of epidermal stem cells around the wound edge. This article discusses the biological characteristics of DETCs and their roles in the maintenance of epidermal homeostasis and wound healing.


Asunto(s)
Epidermis , Cicatrización de Heridas , Diferenciación Celular , Movimiento Celular , Células Epidérmicas , Queratinocitos
15.
Mol Med Rep ; 23(2): 1, 2021 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-33655326

RESUMEN

Angiotensin II (AngII) is a central signaling molecule of the renin­angiotensin system that serves a vital role in myocardial fibrosis (MF). The present study aimed to investigate the effects of matrix metalloproteinase (MMP)3 on MF progression. To induce cellular fibrosis, H9C2 rat myocardial cells were treated with AngII for 24 h. Subsequently, cells were treated with levocarnitine, or transfected with small interfering (si)RNA­negative control or siRNA­MMP3 (1/2/3). Cell viability, apoptosis and migration were assessed by performing Cell Counting Kit­8, flow cytometry and Transwell assays, respectively. Reverse transcription­quantitative PCR (RT­qPCR) and western blotting were performed to determine the expression levels of MF biomarkers, including disease­, apoptosis­ and oxidative stress­related genes. Compared with the control group, AngII significantly inhibited H9C2 cell viability and migration, and significantly increased H9C2 cell apoptosis (P<0.05). However, compared with AngII­treated H9C2 cells, MMP3 knockdown significantly inhibited fibrotic H9C2 cell viability and migration, but increased fibrotic H9C2 cell apoptosis (P<0.05). The RT­qPCR results demonstrated that MMP3 knockdown significantly downregulated the expression levels of AXL receptor tyrosine kinase, AngII receptor type 1, α­smooth muscle actin and Collagen I in AngII­treated H9C2 cells (P<0.05). Moreover, compared with AngII­treated cells, MMP3 knockdown significantly decreased Bcl­2 expression levels , but significantly increased caspase­3 and p53 expression levels in AngII­treated cells (P<0.05). Additionally, compared with AngII­treated cells, MMP3 knockdown significantly decreased MMP3, MMP9, STAT3, p22Phox and p47Phox expression levels in AngII­treated cells (P<0.05). The present study indicated that MMP3 knockdown altered myocardial fibroblast cell viability, migration and apoptosis by regulating apoptosis­ and oxidative stress­related genes, thus delaying MF progression.


Asunto(s)
Angiotensina II/farmacología , Apoptosis , Fibrosis/genética , Corazón/fisiopatología , Metaloproteinasa 3 de la Matriz/metabolismo , Angiotensina II/metabolismo , Animales , Movimiento Celular , Supervivencia Celular , Células Cultivadas , Fibrosis/inducido químicamente , Fibrosis/metabolismo , Fibrosis/fisiopatología , Corazón/efectos de los fármacos , Miocardio/metabolismo , Ratas , Transducción de Señal
17.
J Transl Med ; 19(1): 99, 2021 03 06.
Artículo en Inglés | MEDLINE | ID: mdl-33676540

RESUMEN

BACKGROUND: Glioma, the most common primary brain tumor, account Preparing figures for 30 to 40% of all intracranial tumors. Herein, we aimed to study the effects of M2 macrophage-derived exosomal microRNAs (miRNAs) on glioma cells. METHODS: First, we identified seven differentially expressed miRNAs in infiltrating macrophages and detected the expression of these seven miRNAs in M2 macrophages. We then selected hsa-miR-15a-5p (miR-15a) and hsa-miR-92a-3p (miR-92a) for follow-up studies, and confirmed that miR-15a and miR-92a were under-expressed in M2 macrophage exosomes. Subsequently, we demonstrated that M2 macrophage-derived exosomes promoted migration and invasion of glioma cells, while exosomal miR-15a and miR-92a had the opposite effects on glioma cells. Next, we performed the target gene prediction in four databases and conducted target gene validation by qRT-PCR, western blot and dual luciferase reporter gene assays. RESULTS: The results revealed that miR-15a and miR-92a were bound to CCND1 and RAP1B, respectively. Western blot assays demonstrated that interference with the expression of CCND1 or RAP1B reduced the phosphorylation level of AKT and mTOR, indicating that both CCND1 and RAP1B can activate the PI3K/AKT/mTOR signaling pathway. CONCLUSION: Collectively, these findings indicate that M2 macrophage-derived exosomal miR-15a and miR-92a inhibit cell migration and invasion of glioma cells through PI3K/AKT/mTOR signaling pathway.


Asunto(s)
Neoplasias Encefálicas/metabolismo , Exosomas/metabolismo , Glioma/metabolismo , Macrófagos/metabolismo , MicroARNs/metabolismo , Transducción de Señal , Neoplasias Encefálicas/patología , Línea Celular Tumoral , Movimiento Celular , Biología Computacional , Ciclina D1/metabolismo , Glioma/patología , Humanos , Microscopía Electrónica de Transmisión , Nanopartículas/química , Invasividad Neoplásica , Fosforilación , Proteínas Proto-Oncogénicas c-akt/metabolismo , Reacción en Cadena en Tiempo Real de la Polimerasa , Células THP-1 , Serina-Treonina Quinasas TOR/metabolismo , Proteínas de Unión al GTP rap/metabolismo
18.
J Vis Exp ; (168)2021 02 13.
Artículo en Inglés | MEDLINE | ID: mdl-33645552

RESUMEN

Development of the palate is a dynamic process, which involves vertical growth of bilateral palatal shelves next to the tongue followed by elevation and fusion above the tongue. Defects in this process lead to cleft palate, a common birth defect. Recent studies have shown that palatal shelf elevation involves a remodeling process that transforms the orientation of the shelf from a vertical to a horizontal one. The role of the palatal shelf mesenchymal cells in this dynamic remodeling has been difficult to study. Time-lapse-imaging-based quantitative analysis has been recently used to show that primary mouse embryonic palatal mesenchymal (MEPM) cells can self-organize into a collective movement. Quantitative analyses could identify differences in mutant MEPM cells from a mouse model with palate elevation defects. This paper describes methods to isolate and culture MEPM cells from E13.5 embryos-specifically for time-lapse imaging-and to determine various cellular attributes of collective movement, including measures for stream formation, shape alignment, and persistence of direction. It posits that MEPM cells can serve as a proxy model for studying the role of palatal shelf mesenchyme during the dynamic process of elevation. These quantitative methods will allow investigators in the craniofacial field to assess and compare collective movement attributes in control and mutant cells, which will augment the understanding of mesenchymal remodeling during palatal shelf elevation. Furthermore, MEPM cells provide a rare mesenchymal cell model for investigation of collective cell movement in general.


Asunto(s)
Movimiento Celular , Separación Celular/métodos , Embrión de Mamíferos/citología , Mesodermo/citología , Paladar (Hueso)/citología , Imagen de Lapso de Tiempo , Animales , Rastreo Celular , Células Cultivadas , Criopreservación , Modelos Animales de Enfermedad , Disección , Femenino , Ratones , Cicatrización de Heridas
19.
Biomed Environ Sci ; 34(2): 139-151, 2021 Feb 20.
Artículo en Inglés | MEDLINE | ID: mdl-33685573

RESUMEN

Objective: The underlying mechanism of Ezrin in ovarian cancer (OVCA) is far from being understood. Therefore, this study aimed to assess the role of Ezrin in OVCA cells (SKOV3 and CaOV3) and investigate the associated molecular mechanisms. Methods: We performed Western blotting, reverse transcription-quantitative polymerase chain reaction, MTT, cell colony, cell wound healing, transwell migration and invasion, RhoA and Rac active pull down assays, and confocal immunofluorescence experiments to evaluate the functions and molecular mechanisms of Ezrin overexpression or knockdown in the proliferation and metastasis of OVCA cells. Results: The ectopic expression of Ezrin significantly increased cell proliferation, invasiveness, and epithelial-mesenchymal transition (EMT) in OVCA cells. By contrast, the knockdown of endogenous Ezrin prevented OVCA cell proliferation, invasiveness, and EMT. Lastly, we observed that Ezrin can positively regulate the active forms of RhoA rather than Rac-1 in OVCA cells, thereby promoting robust stress fiber formation. Conclusion: Our results indicated that Ezrin regulates OVCA cell proliferation and invasiveness by modulating EMT and induces actin stress fiber formation by regulating Rho-GTPase activity, which provides novel insights into the treatment of the OVCA.


Asunto(s)
Proteínas del Citoesqueleto/metabolismo , Neoplasias Ováricas/patología , Línea Celular Tumoral , Movimiento Celular , Proliferación Celular , Proteínas del Citoesqueleto/genética , Transición Epitelial-Mesenquimal , Femenino , Regulación Neoplásica de la Expresión Génica , Humanos , Invasividad Neoplásica , Neoplasias Ováricas/genética , Neoplasias Ováricas/metabolismo , Fibras de Estrés/genética , Fibras de Estrés/metabolismo , Proteína de Unión al GTP rhoA/genética , Proteína de Unión al GTP rhoA/metabolismo
20.
Medicine (Baltimore) ; 100(11): e20722, 2021 Mar 19.
Artículo en Inglés | MEDLINE | ID: mdl-33725920

RESUMEN

ABSTRACT: Bladder cancer-associated transcript 1 (BLACAT1) is one of the most common cancer-associated long non-coding RNAs (lncRNAs), which has been reported as a tumor promotor in several malignancies. Previously, BLACAT1 was found to be overexpressed in glioma tissues and cell lines. Functional assays determined that BLACAT1 promoted glioma cell proliferation, migration, invasion and epithelial-mesenchymal transition, suggesting that BLACAT1 might serve as an oncogene in glioma. In the present study, we aimed to investigate its clinical significance and prognostic value in glioma patients.A total of 137 paired glioma tissue samples and adjacent normal brain tissue samples were collected from 137 glioma patients who underwent surgery from May 2014 to February 2019. The Student t test was applied to determine the statistical significance of the observed differences between 2 groups. Survival curves were constructed and differences among groups were calculated using the Kaplan-Meier method.The relative expression of BLACAT1 in glioma samples was significantly higher than that of matched normal tissues (P < .001). The expression level of tissue BLACAT1 was statistically correlated with tumor size (P = .04), Karnofsky Performance Status (KPS) (P = .006), and WHO grade (P = .017). Kaplan-Meier analysis with the log-rank test revealed that BLACAT1 up-regulation was correlated with shorter overall survival time of patients with glioma (Log Rank test, P = .012). In multivariate Cox analysis, BLACAT1 expression was found to be an independent prognostic factor for overall survival in patients with glioma (HR = 2.739; 95% CI: 1.785-8.229; P = .035). Our study demonstrates that up-regulation of BLACAT1 is able to predict aggressive clinicopathologic characteristics and poor prognosis of glioma patients. These findings may have significant implications for potential treatment options and prognosis for patients with glioma.


Asunto(s)
Regulación Neoplásica de la Expresión Génica/genética , Glioma/genética , ARN Largo no Codificante/metabolismo , Regulación hacia Arriba/genética , Encéfalo/metabolismo , Estudios de Casos y Controles , Movimiento Celular/genética , Proliferación Celular/genética , Transición Epitelial-Mesenquimal/genética , Femenino , Glioma/mortalidad , Humanos , Estimación de Kaplan-Meier , Estado de Ejecución de Karnofsky , Masculino , Persona de Mediana Edad , Invasividad Neoplásica/genética , Pronóstico , Modelos de Riesgos Proporcionales
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