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BACKGROUND & AIMS: Noninvasive and accurate methods are needed to identify patients with clinically significant portal hypertension (CSPH). We investigated the ability of deep convolutional neural network (CNN) analysis of computed tomography (CT) or magnetic resonance (MR) to identify patients with CSPH. METHODS: We collected liver and spleen images from patients who underwent contrast-enhanced CT or MR analysis within 14 days of transjugular catheterization for hepatic venous pressure gradient measurement. The CT cohort comprised participants with cirrhosis in the CHESS1701 study, performed at 4 university hospitals in China from August 2016 through September 2017. The MR cohort comprised participants with cirrhosis in the CHESS1802 study, performed at 8 university hospitals in China and 1 in Turkey from December 2018 through April 2019. Patients with CSPH were identified as those with a hepatic venous pressure gradient of 10 mm Hg or higher. In total, we analyzed 10,014 liver images and 899 spleen images collected from 679 participants who underwent CT analysis, and 45,554 liver and spleen images from 271 participants who underwent MR analysis. For each cohort, participants were shuffled and then sampled randomly and equiprobably for 6 times into training, validation, and test data sets (ratio, 3:1:1). Therefore, a total of 6 deep CNN models for each cohort were developed for identification of CSPH. RESULTS: The CT-based CNN analysis identified patients with CSPH with an area under the receiver operating characteristic curve (AUC) value of 0.998 in the training set (95% CI, 0.996-1.000), an AUC of 0.912 in the validation set (95% CI, 0.854-0.971), and an AUC of 0.933 (95% CI, 0.883-0.984) in the test data sets. The MR-based CNN analysis identified patients with CSPH with an AUC of 1.000 in the training set (95% CI, 0.999-1.000), an AUC of 0.924 in the validation set (95% CI, 0.833-1.000), and an AUC of 0.940 in the test data set (95% CI, 0.880-0.999). When the model development procedures were repeated 6 times, AUC values for all CNN analyses were 0.888 or greater, with no significant differences between rounds (P > .05). CONCLUSIONS: We developed a deep CNN to analyze CT or MR images of liver and spleen from patients with cirrhosis that identifies patients with CSPH with an AUC value of 0.9. This provides a noninvasive and rapid method for detection of CSPH (ClincialTrials.gov numbers: NCT03138915 and NCT03766880).
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Hipertensão Portal , Humanos , Hipertensão Portal/complicações , Hipertensão Portal/diagnóstico , Cirrose Hepática/complicações , Cirrose Hepática/diagnóstico , Redes Neurais de Computação , Pressão na Veia PortaRESUMO
BACKGROUND: Cancer cells survival depends on glucose metabolism and ATP. Inhibiting glucose metabolism is a possible anticancer treatment. The phosphorylation of 2-deoxy-D-glucose (2-DG), which is a glycogen analogue, seriously affects the normal glycometabolism phosphorylation process, leading to ATP consumption. Studies showed that 2-DG could regulate RIP and c-FLIP. This paper aimed to investigate the effect of 2-DG on RIP and c-FLIP expression in HepG2 and Hep3B cells, further illustrating the effect and mechanism of 2-DG regulating RIP and c-FLIP expression on liver cancer cell apoptosis induced by TRAIL. MATERIAL AND METHODS: RIP and c-FLIP gene silencing HepG2 and Hep3B cell models were established by siRNA and detected by Western blot. Cell viability was determined by MTT and apoptosis rate was measured by flow cytometry. JC-1 fluorescent probe was used to test mitochondrial membrane potential. RESULTS: 2-DG or TRAIL alone significantly reduced HepG2 and Hep3B cell survival rate and promoted apoptosis. Compared with the single TRAIL treatment group, the combination of 2-DG and TRAIL could reduce cell survival rate, increase apoptosis rate, and decease mitochondrial membrane potential, which is dependent on Caspases. 2-DG can inhibit RIP and c-FLIP expression, leading to increased TRAIL-induced HepG2 and Hep3B cells apoptosis. CONCLUSIONS: 2-DG can down-regulate RIP and c-FLIP expression, and change Caspases activities to increase the liver cancer cell apoptosis induced by TRAIL.
Assuntos
Apoptose , Proteína Reguladora de Apoptosis Semelhante a CASP8 e FADD/metabolismo , Desoxiglucose/química , Neoplasias Hepáticas/patologia , Proteína Serina-Treonina Quinases de Interação com Receptores/metabolismo , Ligante Indutor de Apoptose Relacionado a TNF/metabolismo , Trifosfato de Adenosina/química , Citometria de Fluxo , Inativação Gênica , Células Hep G2 , Humanos , Neoplasias Hepáticas/tratamento farmacológico , Neoplasias Hepáticas/metabolismo , Potenciais da Membrana , Mitocôndrias/patologia , RNA Interferente Pequeno/metabolismoRESUMO
Inflammation plays an essential role in the development liver fibrosis.The Cyclic guanosine monophosphate-adenosine monophosphate synthase (cGAS) is a central cytoplasmic DNA sensor which can recognize cytoplasmic DNA, known to trigger stimulator of interferon genes (STING) and downstream proinflammatory factors. Here, we investigated the role of cGAS-STING signaling pathway in the pathogenesis of liver fibrosis.Differentially expressed genes (DEGs) in human liver tissue were identified using RNA-Seq analysis. As models of liver fibrosis, chronic Carbon tetrachloride (CCl4) exposure were applied in cGAS-knockout mice. LX-2 cells were co-cultured with human liver sinusoidal endothelial cells (LSECs) to explore the underlying mechanisms of hepatic sinusoidal microthrombosis in an inflammatory microenvironment. The endoscopic ultrasound-guided portal vein pressure gradient (EUS-PPG) method was used to analyze the associations between hepatic sinusoidal microthrombosis and PPG in patients with liver fibrosis and portal hypertension (PTH). The RNA-seq analysis results showed that DEGs were enriched in inflammation and endothelial cell activation. The upregulation of the cGAS-STING signaling exacerbated liver fibrosis and intrahepatic inflammation. It also exacerbated LSECs impairment and increased the contribution of hepatic sinusoidal microthrombosis to liver fibrosis in vivo and in vitro. Prothrombotic mediators and proinflammatory factors were associated with PPG in patients with liver fibrosis and portal hypertension. Therefore, activating cGAS-STING signaling pathway promotes liver fibrosis and hepatic sinusoidal microthrombosis, which may lead to increased portal vein pressure.
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Células Endoteliais , Hipertensão Portal , Animais , Camundongos , Humanos , Cirrose Hepática , Transdução de Sinais , Cromogranina A , DNA , InflamaçãoRESUMO
The hepatic venous pressure gradient (HVPG) is the gold standard for cirrhotic portal hypertension (PHT), but it is invasive and specialized. Alternative non-invasive techniques are needed to assess the hepatic venous pressure gradient (HVPG). Here, we develop an auto-machine-learning CT radiomics HVPG quantitative model (aHVPG), and then we validate the model in internal and external test datasets by the area under the receiver operating characteristic curves (AUCs) for HVPG stages (≥10, ≥12, ≥16, and ≥20 mm Hg) and compare the model with imaging- and serum-based tools. The final aHVPG model achieves AUCs over 0.80 and outperforms other non-invasive tools for assessing HVPG. The model shows performance improvement in identifying the severity of PHT, which may help non-invasive HVPG primary prophylaxis when transjugular HVPG measurements are not available.
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Inteligência Artificial , Hipertensão Portal , Diagnóstico por Imagem , Humanos , Hipertensão Portal/diagnóstico por imagem , Cirrose Hepática/complicações , Pressão na Veia PortaRESUMO
Human amniotic epithelial cells (hAECs) are a recently identified type of stem cell. Thanks to their ready availability and the lower risk of teratoma formation, hAECs have been studied and tested for a variety of human disease treatments and tissue reconstruction efforts. This aim of this study was to establish a stable tracking system to further monitor hAECs in vivo after transplantation. hAECs were isolated from the placentas of patients who visited the Hunan Province Maternity and Child Care Hospitals between Jan 2008 and Jan 2009. Using the classic transfection/infection technique, we successfully introduced green fluorescent protein (GFP) into cultured hAECs with an adeno-associated virus (AAV) vector. The initial preparation of the AAV-GFP virus stock was titrated using HT1081 cells, and further used for the infection of hAECs. GFP(+) hAECs preserve the capacity of differentiation into hepatocyte-like cells with the expression of cytokeratin-18 (CK18) and albumin (ALB). AAV-GFP virus-infected hAECs were transplanted through the spleen into severe combined immune deficiency (SCID) mice via hepatectomy. Four weeks later, the GFP and human albumin expressions were examined in multiple organs through immunofluorescence staining. In culture, over 50% of the hAECs were GFP-positive 3 days after infection. Following transplantation, AAV-GFP-infected hAECs survived and continued to express GFP in the host for up to 4 weeks. These cells were primarily found in the spleen and liver, expressing human albumin. This study provides a feasible and stable system to track hAECs. It may prove useful to further identify their biological characteristics after transplantation and to elucidate their beneficial roles for therapeutic purposes.
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Âmnio/citologia , Rastreamento de Células , Dependovirus/genética , Células Epiteliais/metabolismo , Vetores Genéticos/genética , Proteínas de Fluorescência Verde/metabolismo , Animais , Diferenciação Celular , Células Epiteliais/citologia , Feminino , Vetores Genéticos/metabolismo , Proteínas de Fluorescência Verde/genética , Humanos , Hospedeiro Imunocomprometido , Queratina-18/metabolismo , Camundongos , Gravidez , Albumina Sérica/genética , Albumina Sérica/metabolismoRESUMO
OBJECTIVE: To evaluate the in vitro differentiation of human amniotic epithelial cells (hAECs ) into hepatocyte-like cells. METHODS: Combined approach of dexamethasone, HGF, IGF and other cytokines were used to induce the differentiation of hAECs into hepatocyte-like cells. The induction lasted 2 weeks. During the induction, the expression of albumin ALB, CYP1A1, CYP1A2, IGFR, c-met and key functional genes related to liver cells as well as transcription factors HNF3, HNF4 and C/EBPa were monitored by RT-PCR. Time dependent changes of the surface marker colony ALB, AFP and CK18 were analyzed by cell flow cytometry. RESULTS: After the 2 week induction, the expressions of liver hepatocyte-like cell functional genes such as albumin, CYP1A1, CYP1A2, c-met, and transcription factors such as HNF3, HNF4, C/EBPa and HNF1 were observed. Six days after the induction, hAECs mainly were stained AFP+, and the positive rate was (15.1 ± 2.1)%. While 10 days after the induction, part of the hAECs showed AFP+/ALB+ (6.5 ± 1.4)%; and on 14th day, hAECs only showed ALB+, and the rate was (13.9 ± 2.3)%. ALB+ cell increase indicated a gradual functional maturation from the hAECs to hepatocyte-like cells. Similaritly, the number of CK18+ cells in the whole population was also increased: On 10th day, the rate was (16.1 ± 1.2)%; on 14th day, that was (21.3 ± 4.6)%, which proved the above hypothesis of the trandifferentiation. By extending the induction time, the expression of functional genes increased gradually, and a maturing process of hAECs was detected by cell surface markers. CONCLUSION: The differentiation of hAECs induced in vitro has the characteristics of hepatocyte-like cells.
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Âmnio/citologia , Diferenciação Celular , Células Epiteliais/citologia , Hepatócitos/citologia , Células Cultivadas , Dexametasona/farmacologia , Fator de Crescimento de Hepatócito/farmacologia , Humanos , Somatomedinas/farmacologiaRESUMO
Jaundice occurs in 19%-40% of the hepatocellular carcinoma (HCC) patients. HCC associated jaundice may be divided into hepatocellular and icteric types in terms of its underlying pathophysiology. The jaundice of icteric type is caused by obstruction of the bile duct through cancer embolus, blood clot, biliary sludge, tumor compression or infiltration. Jaundice and epigastric discomforts are the main clinical manifestations. In the present case, severe acute pancreatitis and acute cholangitis presenting as initial complaints of icteric type HCC were quite rare. A tumor located at the central lobe of the liver and a cancer embolus at the lower part of the common bile duct (CBD) were detected by CT scan. Curative resection of HCC with CBD exploration eradicated both the tumor and the embolus, and no recurrence was found after a 36 month follow-up.
Assuntos
Carcinoma Hepatocelular/diagnóstico , Colangite/diagnóstico , Neoplasias Hepáticas/diagnóstico , Pancreatite/diagnóstico , Carcinoma Hepatocelular/complicações , Carcinoma Hepatocelular/cirurgia , Colangite/etiologia , Diagnóstico Diferencial , Seguimentos , Humanos , Neoplasias Hepáticas/complicações , Neoplasias Hepáticas/cirurgia , Masculino , Pessoa de Meia-Idade , Pancreatite/etiologiaRESUMO
OBJECTIVE: To determine the therapeutic effect of laparoscopic splenectomy, perisoph-agogastric devascularization, and endoscopic variceal ligation (EVL) on patients with portal hypertension. METHODS: We randomly divided 105 patients into 3 groups: 40 had endoscopic band ligation (the ligation group), 35 had splenectomy and perisoph-agogastric devascularization (the laparotomy group), and the other 30 had laparoscopic splenectomy, perisoph-agogastric devascularization and endoscopic variceal ligation (the combination group). Blood samples were analyzed preoperatively and postoperatively on day 1,3,and 7,including alanine aminotransferase(ALT),aspartate aminotransferase(AST),total bilirubin(TBIL),and directed bilirubin(DBIL). The length of stay, blood loss, operation time, anal exhaust time, azygos vein diameter, blood flow velocity and blood flow, recurrence of esophageal varices and rehaemorrhagia were compared. RESULTS: Between the combination group and the laparotomy group, the serum levels of TbIL and Dbil had difference on 1st postoperative day(P<0.05). AST had difference on 7th postoperative day(P<0.05). The length of stay, blood loss, operation time, and anal exhaust time had significant difference(P<0.05). Among the combination group, the laparotomy group and the ligation group, the azygos vein blood flow before and after the treatment, recurrence of esophageal varices and rehaemorrhagia had no difference(P<0.05). CONCLUSION: Laparoscopic splenectomy, perisoph-agogastric devascularization and endoscopic variceal ligation have less trauma, lower recurrence rate, fewer complications and rapid recovery, and may reduce the azygous vein blood flow. It can be used safely for portal hypertension.
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Endoscopia/métodos , Varizes Esofágicas e Gástricas/cirurgia , Hipertensão Portal/cirurgia , Laparoscopia/métodos , Adulto , Varizes Esofágicas e Gástricas/complicações , Feminino , Humanos , Hipertensão Portal/complicações , Ligadura/métodos , Masculino , Pessoa de Meia-Idade , Esplenectomia/métodosRESUMO
Liver fibrosis is a common response to chronic liver injury, ultimately leading to cirrhosis. The activation of hepatic stellate cells (HSCs) plays a dominant role in liver fibrosis. The regulatory roles of long noncoding RNAs (lncRNAs) in multiple human diseases have been observed. This study was dedicated to investigating the regulatory effects of the lncRNA nuclear paraspeckle assembly transcript 1 (Neat1) on liver fibrosis and HSC activation. Upregulation of Neat1 and cytohesin 3 (Cyth3) and downregulation of miR-148a-3p and miR-22-3p were observed in mouse fibrotic liver tissues. Knockdown of Neat1 or Cyth3 attenuated liver fibrosis and collagen deposition in vivo and the activation of HSCs in vitro. An miR-148a-3p and miR-22-3p inhibitor facilitated HSC activation and collagen fiber expression. Neat1 directly targeted miR-148a-3p and miR-22-3p to modulate Cyth3 expression. Knockdown of Neat1 inhibited Cyth3 expression via the competing endogenous RNA (ceRNA) mechanism of sponging miR-148a-3p and miR-22-3p to regulate liver fibrosis and HSC activation. The ceRNA regulatory network may promote a better understanding of liver fibrogenesis, contribute to an original agreement of liver fibrosis etiopathogenesis and provide insights into the development of a novel domain of lncRNA-directed therapy against liver fibrosis.
Assuntos
Cirrose Hepática/metabolismo , MicroRNAs/metabolismo , RNA Longo não Codificante/biossíntese , Receptores Citoplasmáticos e Nucleares/biossíntese , Regulação para Cima/fisiologia , Animais , Tetracloreto de Carbono/toxicidade , Linhagem Celular , Progressão da Doença , Células Estreladas do Fígado/efeitos dos fármacos , Células Estreladas do Fígado/metabolismo , Células Estreladas do Fígado/patologia , Cirrose Hepática/induzido quimicamente , Cirrose Hepática/patologia , Masculino , Camundongos , Camundongos Endogâmicos C57BL , RNA Longo não Codificante/antagonistas & inibidores , Receptores Citoplasmáticos e Nucleares/antagonistas & inibidores , Regulação para Cima/efeitos dos fármacosRESUMO
OBJECTIVE: This study was designed to investigate the effect of long non-coding RNA (lncRNA) OIP5-AS1 on cell migration and invasion of gallbladder cancer (GBC) and its specific mechanism. METHODS: The expressions of lncRNA OIP5-AS1 and miR-143-3p in GBC cell lines (GBC-SD, SGC996 and NOZ) and gallbladder epithelial cells (HGBE cells) were measured by qRT-PCR. After loss- and gain-of-function experiments for OIP5-AS1 and miR-143-3p in GBC-SD cells, CCK-8 was applied to examine cell viability, cell scratch assay to measure cell migration, and transwell chamber to inspect cell invasion capacity. The interaction between OIP5-AS1 and miR-143-3p was predicted by StarBase. Then, luciferase reporter gene assay and RNA pull-down were used to verify the targeting relationship between miR-143-3p and OIP5-AS1. RESULTS: OIP5-AS1 was highly expressed and miR-143-3p was downregulated in GBC cell lines, when compared with HGBE cells. Overexpression of OIP5-AS1 or downregulation of miR-143-3p facilitated GBC-SD cell invasion, proliferation and migration, while different expression patterns were found in GBC-SD cells in response to OIP5-AS1 suppression or miR-143-3p overexpression. OIP5-AS1 negatively mediated miR-143-3p. MiR-143-3p upregulation partially reversed the inhibitory effect of OIP5-AS1 knockdown on GBC-SD cell activities. CONCLUSION: LncRNA OIP5-AS1 accelerates the progression of GBC by suppressing miR-143-3p.
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BACKGROUND: The most common malignant tumor of the digestive system is HCC. However, the mechanism and pathogenesis of HCC occurrence and progress are still unknown. LncRNA is closely related to the occurrence and progress of HCC. It is important to investigate the effect and role of lncRNA in HCC. MATERIALS AND METHODS: LncRNA microarray assay was used to screen the differential expression profile of lncRNA. SNHG11, miR-184 and GO2 expression was analyzed by RT-PCR. The ability of SNHG11 to serve as a sponge for miRNA and the fact that miR-184 directly targets mRNA were revealed by dual luciferase assay and RIP. Apoptosis and autophagy related proteins were detected by Western blot. Cell proliferation, invasion, migration, and apoptosis were detected by CCK-8 assay, wound healing assay, transwell assay, and flow cytometry. RESULTS: LncRNA microarray assay and RT-PCR results revealed that the expression of SNHG11 was increased in HCC tumor tissues and also upregulated in HCC cells. SNHG11 had a connection with poor survival rate in HCC. In addition, dual luciferase assay and RIP results revealed that SNHG11 serves as a sponge for miR-184 and miR-184 directly targets AGO2. Pearson correlation analysis showed that SNHG11 with miR-184 and miR-184 with AGO2 were negative correlations, and SNHG11 with AGO2 was a positive correlation. Cell function assay and Western blot showed SNHG4/miR-184/AGO2 regulatory loop was critical for HCC cell proliferation, migration, apoptosis, and autophagy. CONCLUSION: Our study demonstrated that the expression of SNHG11 is higher in HCC; moreover, SNHG11 promotes proliferation, migration, apoptosis, and autophagy by regulating AGO2 via miR-184 in HCC. Our verification of the role of SNHG11 may provide a novel biomarker for the diagnosis, therapy, and prognosis of HCC.
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Hepatocellular carcinoma (HCC) is the fifth most common cancer worldwide. LncRNA small nucleolar RNA host gene 14 (SNHG14) functions as an oncogene in a variety of cancers. However, the role of SNHG14 in HCC remains elusive. The aim of this study is to unravel the functional role and regulatory mechanism of SNHG14 in HCC. A cohort of 40 HCC tumor tissues and paired adjacent normal tissues were collected. Histopathological changes were analyzed by hematoxylin and eosin and immunohistochemistry. qRT-PCR and western blotting were performed to determine the levels of SNHG14, PABPC1, and PTEN signaling molecules. CCK-8, immunofluorescence, and colony formation assays were conducted to monitor cell proliferation. Wound healing and tube formation assays were employed to determine cell migration and angiogenesis. ChIP assay was performed to investigate the enrichment of H3K27 acetylation in PABPC1 promoter. Xenograft mice model was constructed to further verify the SNHG14/PABPC1 axis in vivo. SNHG14 was highly expressed in HCC tissues and cells, which promoted cell proliferation, migration, and angiogenesis in Hep3B and HepG2 cells. PABPC1 functioned as a downstream effector of SNHG14. SNHG14 dramatically induced upregulation of PABPC1 via H3K27 acetylation. In addition, SNHG14/PABPC1 promoted cell proliferation and angiogenesis via PTEN signaling pathway in vitro and in vivo. SNHG14 promoted cell proliferation and angiogenesis via upregulating PABPC1 through H3K27 acetylation and modulating PTEN signaling in the tumorigenesis of HCC.
Assuntos
Carcinoma Hepatocelular/genética , Proteína I de Ligação a Poli(A)/genética , RNA Nucleolar Pequeno/genética , Acetilação , Adulto , Idoso , Idoso de 80 Anos ou mais , Apoptose/genética , Linhagem Celular Tumoral , Movimento Celular/genética , Proliferação de Células/genética , China , Progressão da Doença , Feminino , Regulação Neoplásica da Expressão Gênica/genética , Células Hep G2 , Histonas/metabolismo , Humanos , Neoplasias Hepáticas/genética , Masculino , MicroRNAs/genética , Pessoa de Meia-Idade , PTEN Fosfo-Hidrolase/genética , PTEN Fosfo-Hidrolase/metabolismo , Proteína I de Ligação a Poli(A)/metabolismo , RNA Longo não Codificante/genética , RNA Nucleolar Pequeno/metabolismo , Proteínas de Ligação a RNA/metabolismo , Transdução de Sinais/genéticaRESUMO
OBJECTIVE: To investigate the effects of over-expression of miR-144 on invasion of SMMC-7721 cells and Toll-like receptor (TLR)/myeloid differentiation factor 88 (MyD88) pathway in hepatocellular carcinoma cells. METHODS: The expressions of miR-144 was examined in normal human hepatocyte line HL-7702 and hepatocarcinoma cell line SMMC-7721 using realtime quantitative PCR (qRT-PCR). SMMC-7721 cells were divided into blank group, miR-144 NC group and miR-144 mimics group, and the expressions of miR-144 in each group were detected with qRT-PCR. Cell count kit-8 (CCK8) was used to assess the survival of SMMC-7721 cells, and the cell invasion was evaluated using Transwell assay. The expressions of matrix metalloproteinase-2 (MMP-2), matrix metalloproteinase-9 (MMP-9) and TLR/MyD88 pathway-related proteins in the cells were detected with Western blotting; the effect of 40 µ mol/L MyD88 inhibitor on TLR/MyD88 pathway-related proteins was examined in SMMC-7721 cells. RESULTS: Compared with normal human hepatocytes, SMMC-7721 cells expressed a significantly lower level of miR-144 (P < 0.05). CCK-8 assay showed that test showed that miR-144 over-expression significantly decreased the cell survival rate (P < 0.05), lowered the number of invasive cells, and decreased the expression of MMP-2 and MMP-9 in SMMC-7721 cells (P < 0.05). The expressions of Toll-like receptor 4 (TLR4), MyD88, phosphorylated nuclear factor-kappa B (pNF-κB) and NF-κB protein decreased significantly in miR-144 mimics group and TJ-M2010-2 group (P < 0.05) and were comparable between the two groups (P > 0.05). CONCLUSIONS: Overexpression of miR-144 decreases SMMC-7721 cell survival and invasion by inhibiting TLR/MyD88 pathway.
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Neoplasias Hepáticas , Linhagem Celular Tumoral , Humanos , Metaloproteinase 2 da Matriz , MicroRNAs , Fator 88 de Diferenciação Mieloide , NF-kappa B , Transdução de Sinais , Receptores Toll-LikeRESUMO
Hepatocellular carcinoma is one of the most common solid tumors in the digestive system. The prognosis of patients with hepatocellular carcinoma is still poor due to the acquisition of multi-drug resistance. TNF Related Apoptosis Inducing Ligand (TRAIL), an attractive anticancer agent, exerts its effect of selectively inducing apoptosis in tumor cells through death receptors and the formation of the downstream death-inducing signaling complex, which activates apical caspases 3/8 and leads to apoptosis. However, hepatocellular carcinoma cells are resistant to TRAIL. Non-coding RNAs, including long non-coding RNAs (lncRNAs) and miRNAs have been regarded as major regulators of normal development and diseases, including cancers. Moreover, lncRNAs and miRNAs have been reported to be associated with multi-drug resistance. In the present study, we investigated the mechanism by which TRAIL resistance of hepatocellular carcinoma is affected from the view of non-coding RNA regulation. We selected and validated candidate miRNAs, miR-24 and miR-221, that regulated caspase 3/8 expression through direct targeting, and thereby affecting TRAIL-induced tumor cell apoptosis TRAIL resistance of hepatocellular carcinoma. In addition, we revealed that CASC2, a well-established tumor suppressive long non-coding RNA, could serve as a "Sponge" of miR-24 and miR-221, thus modulating TRAIL-induced tumor cell apoptosis TRAIL resistance of hepatocellular carcinoma. Taken together, we demonstrated a CASC2/miR-24/miR-221 axis, which can affect the TRAIL resistance of hepatocellular carcinoma through regulating caspase 3/8; through acting as a "Sponge" of miR-24 and miR-221, CASC2 may contribute to improving hepatocellular carcinoma TRAIL resistance, and finally promoting the treatment efficiency of TRAIL-based therapies.
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Carcinoma Hepatocelular/patologia , Caspase 3/metabolismo , Caspase 8/metabolismo , Resistencia a Medicamentos Antineoplásicos , MicroRNAs/metabolismo , Ligante Indutor de Apoptose Relacionado a TNF/farmacologia , Proteínas Supressoras de Tumor/metabolismo , Sequência de Bases , Resistencia a Medicamentos Antineoplásicos/efeitos dos fármacos , Células Hep G2 , Humanos , Neoplasias Hepáticas/patologiaRESUMO
Clinically significant portal hypertension (CSPH) is associated with an incremental risk of esophageal varices and overt clinical decompensations. However, hepatic venous pressure gradient (HVPG) measurement, the gold standard for defining CSPH (HVPG≥10â¯mmâ¯Hg) is invasive and therefore not suitable for routine clinical practice. This study aims to develop and validate a radiomics-based model as a noninvasive method for accurate detection of CSPH in cirrhosis. The prospective multicenter diagnostic trial (CHESS1701, ClinicalTrials.gov identifier: NCT03138915) involved 385 patients with cirrhosis from five liver centers in China between August 2016 and September 2017. Patients who had both HVPG measurement and contrast-enhanced CT within 14â¯days prior to the catheterization were collected. The noninvasive radiomics model, termed rHVPG for CSPH was developed based on CT images in a training cohort consisted of 222 consecutive patients and the diagnostic performance was prospectively assessed in 163 consecutive patients in four external validation cohorts. rHVPG showed a good performance in detection of CSPH with a C-index of 0·849 (95%CI: 0·786-0·911). Application of rHVPG in four external prospective validation cohorts still gave excellent performance with the C-index of 0·889 (95%CI: 0·752-1·000, 0·800 (95%CI: 0·614-0·986), 0·917 (95%CI: 0·772-1·000), and 0·827 (95%CI: 0·618-1·000), respectively. Intraclass correlation coefficients for inter- and intra-observer agreement were 0·92-0·99 and 0·97-0·99, respectively. A radiomics signature was developed and prospectively validated as an accurate method for noninvasive detection of CSPH in cirrhosis. The tool of rHVPG assessment can facilitate the identification of CSPH rapidly when invasive transjugular procedure is not available.
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Biomarcadores , Hipertensão Portal/diagnóstico por imagem , Hipertensão Portal/etiologia , Cirrose Hepática/complicações , Cirrose Hepática/diagnóstico por imagem , Tomografia Computadorizada por Raios X , Feminino , Humanos , Hipertensão Portal/sangue , Processamento de Imagem Assistida por Computador , Cirrose Hepática/sangue , Masculino , Variações Dependentes do Observador , Curva ROC , Reprodutibilidade dos Testes , Tomografia Computadorizada por Raios X/métodos , Tomografia Computadorizada por Raios X/normasAssuntos
Âmnio , Células Cultivadas , Diferenciação Celular , Células Epiteliais , Hepatócitos , HumanosRESUMO
OBJECTIVE: To investigate variations of plasma endothelin (ET) and its clinical significance in portal hypertensive patients with esophageal variceal hemorrhage. METHODS: Sixty-six patients with portal hypertension were randomly divided into 2 groups. Group I (32 patients) received general therapy and Group II (34 patients) received general therapy and UTI after hemorrhage. The plasma ET concentration and liver function were determined at 1, 2, 4, 7, 10, and 14 d after the hemorrhage. Another 20 patients without the hemorrhage were elected as the control group. RESULTS: At 7 and 14 d after the hemorrhage, the levels of TBIL, ALT and AST were elevated at first and then decreased in Groups I and II. The decrease of TBIL, ALT and AST levels was significantly faster in Group II than in Group I (P < 0.05, P < 0.01, P < 0.05, respectively) on 14 d after the hemorrhage. At 1 d after the hemorrhage the ET concentration was markedly increased in Group I and II as compared with the control group (P < 0.01). Then it was gradually decreased on 10 d after the hemorrhage. The ET concentration in Group II was decreased more rapidly than that in Group I on 2, 4 and 7 d after the hemorrhage (P < 0.05; P < 0.01; P < 0.05, respectively). The ET concentration was positively correlated to TBIL levels in groups I and II (r = 0.734, P < 0.01). And the decreased index of ET concentration was negatively correlated to the increased index of TBIL (r = -0.486, P < 0.05). CONCLUSION: The increased plasma ET in portal hypertensive patients with hemorrhage may contribute to liver injury. UTI can protect the liver function by inhibiting ALT, AST, TBIL and ET level.
Assuntos
Endotelina-1/sangue , Varizes Esofágicas e Gástricas/etiologia , Glicoproteínas/uso terapêutico , Hipertensão Portal/complicações , Adulto , Idoso , Varizes Esofágicas e Gástricas/sangue , Feminino , Humanos , Hipertensão Portal/sangue , Falência Hepática/prevenção & controle , Masculino , Pessoa de Meia-Idade , Inibidores da Tripsina/uso terapêuticoRESUMO
OBJECTIVE: To observe the survival, migration, and effect of human amniotic epithelial cells (hAECs) on hepatc fibrosis in immune rats so as to provide the experimental theory for the clinical treatment with hAECs. METHODS: Sixty-four 10-week-old male Sprague Dawley rats (weighing, 220-280 g) were randomly divided into 4 groups, sixteen rats in each group. Rat hepatic fibrosis model was induced in groups A, B, and C; hepatic fibrosis rats were injected with 4 x 10(6) hAECs in group A, and with normal saline in group B, and no treatment was given in group C; group D served as control group. After 2 weeks of transplantation, the expression of human Alu gene repeat sequence was detected by DNA-PCR method and human leucocyte antigen G (HLA-G) by immunohistochemical staining in heart, liver, spleen, kidney, lung, and brain in group A, and then the percentage of positive expression was compared between organs except spleen. Semi-quantitative analysis was done for liver fibrosis with HE staining according to Chevallier semi-quantitative histological liver fibrosis scoring system, and immunohistochemical staining for TGF-ß1 was used to record immunohistochemical score (ISH), the concentrations of aspartate transaminase (AST), alanine aminotransferase (ALT), and albumin (ALB) were determined to analyze hepatic fibrosis. RESULTS: Alu gene repeat sequence and HLA-G could be detected in liver, heart, brain, lung, and kidney in group A, the percentage of positive expression in the liver was significantly higher than that in the other organs (P < 0.05). The histological semi-quantitative score of group A (10.47 ± 3.20) was significantly lower than that of groups B and C [(13.84 ± 3.46) and (13.85 ± 3.16)](P < 0.05), but no significant difference was found between groups B and C (P > 0.05). The ISH scores in groups A, B, C, and D were 3.60 ± 1.50, 5.38 ± 2.60, 5.50 ± 2.40, and 1.87 ± 1.36, respectively; groups A, B, and C were significantly higher than group D, and group A was significantly lower than groups B and C (P < 0.05), but there was no significant difference between groups B and C (P > 0.05). The concentrations of ALT and AST in groups A, B, and C were significantly higher than those in group D, and group A was significantly lower than groups B and C (P < 0.05), but there was no significant difference between groups B and C (P > 0.05). The concentration of ALB in groups A, B, and C was significantly lower than that in group D, and group A was significantly higher than groups B and C (P<0.05), but there was no significant difference between groups B and C (P > 0.05). CONCLUSION: hAECs can survive in immune rats by intrasplenic transplantation and migrate to liver, heart, brain, lung, and kidney, and the liver shows the largest migration. The transplantation of hAECs in immune rat with cirrhosis can alleviate hepatic fibrosis and improve the serum indexes of liver function.
Assuntos
Âmnio/patologia , Cirrose Hepática Experimental/metabolismo , Cirrose Hepática Experimental/patologia , Fígado/patologia , Alanina Transaminase/sangue , Alanina Transaminase/metabolismo , Âmnio/metabolismo , Animais , Aspartato Aminotransferases/sangue , Aspartato Aminotransferases/metabolismo , Células Epiteliais/metabolismo , Fibrose/terapia , Humanos , Fígado/metabolismo , Masculino , Ratos , Ratos Sprague-Dawley , Fator de Crescimento Transformador beta1RESUMO
The incidence and mortality of liver cancer increased year by year. Our country presents high incidence of liver cancer. MicroRNAs have tissue sensitivity as tumor biomarkers that play a role by promoting tumor growth as oncogenes or inhibit malignant cell growth as tumor suppressor genes. Studies showed that miR-15b abnormal expression in the tumor and can be treated as one of the tumor molecular markers. However, miR-15b expression and role in the liver cancer cells have not been elucidated. This study intended to explore the mechanism of miR-15b effect on liver cancer occurrence and development. Liver cancer cell line HepG2 was transfected with miR-15b mimic or inhibitor. Real-time PCR was applied to detect miR-15b expression. MTT was used to test cell proliferation. Transwell assay was performed to determine cell invasive ability. Real-time PCR and Western blot were used to detect BCL2 expression. MiR-15b mimic transfection promoted miR-15b overexpression and inhibited HepG2 cell proliferation significantly (P < 0.05). MiR-15b overexpression downregulated BCL2 mRNA and protein expression obviously (P < 0.05). On the contrary, miR-15b inhibitor transfection markedly reduced miR-15b expression in liver cancer cells (P < 0.05), promoted tumor cell proliferation, and increased BCL2 mRNA and protein expression. MiR-15b expression changes did not affect cell invasion (P > 0.05). MiR-15b can inhibit HepG2 cell proliferation and down-regulate BCL2 mRNA and protein expression.
Assuntos
Biomarcadores Tumorais/metabolismo , Carcinoma Hepatocelular/metabolismo , Proliferação de Células , Neoplasias Hepáticas/metabolismo , MicroRNAs/metabolismo , Proteínas Proto-Oncogênicas c-bcl-2/metabolismo , Biomarcadores Tumorais/genética , Carcinoma Hepatocelular/genética , Carcinoma Hepatocelular/patologia , Movimento Celular , Regulação Neoplásica da Expressão Gênica , Células Hep G2 , Humanos , Neoplasias Hepáticas/genética , Neoplasias Hepáticas/patologia , MicroRNAs/genética , Invasividade Neoplásica , Proteínas Proto-Oncogênicas c-bcl-2/genética , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Transdução de Sinais , TransfecçãoRESUMO
TRAIL (TNF-related apoptosis-inducing ligand) is a member of the tumor necrosis factor superfamily that can induce tumor selective death by up-regulating death receptor 4 (DR4) and DR5 expression. The study aimed to explore the role of RIP and c-FLIP genes in TRAIL induced liver cancer cell HepG2 and Hep3B apoptosis and related mechanism. RIP and c-FLIP silenced HepG2 and Hep3B cell model were established through siRNA. Western blot was applied to test c-FLIP, RIP, DR4, DR5, FADD, Caspase-3/8/9, ERK1/2, and DFF45 protein expression. Caspase-8 kit was used to detect Caspase-8 expression. Flow cytometry was performed to measure cell apoptosis rate. Acid phosphatase method was applied to determine cell cycle. TRAIL had no significant effect on Caspase-3/8/9, DR4, DR5, ERK1/2, and DFF45 protein expression, but up-regulated c-FLIP and RIP protein expression and reduced FADD expression level. After treated by the chemotherapy drug mitomycin and adriamycin, c-FLIP and RIP expression decreased significantly, while FADD increased. After knockout c-FLIP and RIP gene, HepG2 and Hep3B cell apoptosis rate induced by TRAIL increased obviously. Meanwhile, cell subG1 percentage increased markedly and exhibited G1 phase growth retardation. In addition, after two kinds of gene knockout, Caspase-8 was activated and produce Caspase-3 P20 and P24, leading DFF45 appeared DNA fragment P17 and P25. c-FLIP and RIP can inhibit Caspase-8 activation and prompting HepG2 and Hep3B resistant to cell apoptosis induced by TRAIL.