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This retrospective cohort study aims to review our 18-year experience with early hepatic artery thrombosis (e-HAT) following living-donor liver transplantation (LDLT), as well as to assess the feasibility, efficacy and potential risks of endovascular management of e-HAT in the first 48 hours (hrs) post-LDLT. Medical records of 730 patients who underwent LDLT were retrospectively reviewed. In all cases who had developed e-HAT, treatment modalities employed and their outcomes were evaluated. Thirty-one patients developed e-HAT(4.2%). Definite technical success and 1-year survival rates of surgical revascularization[11/31 cases(35.5%)] were 72.7% & 72.7%, whereas those of endovascular therapy[27/31 cases(87.1%)] were 70.4% & 59.3%, respectively. Endovascular therapy was carried out in the first 48hrs post-transplant in 9/31 cases(29%) [definite technical success:88.9%, 1-year survival:55.6%]. Four procedure-related complications were reported in 3 of those 9 cases(33.3%). In conclusion, post-LDLT e-HAT can be treated by surgical revascularization or endovascular therapy, with comparable results. Endovascular management of e-HAT in the first 48hrs post-LDLT appears to be feasible and effective, but is associated with a relatively higher risk of procedure-related complications, compared to surgical revascularization. Hence, it can be reserved as a second-line therapeutic option in certain situations where surgical revascularization is considered futile, potentially too complex, or potentially more risky.
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Procedimentos Endovasculares , Transplante de Fígado , Trombose , Estudos de Viabilidade , Artéria Hepática/cirurgia , Humanos , Transplante de Fígado/efeitos adversos , Doadores Vivos , Estudos Retrospectivos , Trombose/etiologia , Resultado do TratamentoRESUMO
IGF2BP 1, 2 and 3 control the fate of many transcripts. Immunoprecipitation studies demonstrated the IGF2BPs to bind to IGF1R mRNA, and our laboratory has recently shown them to post-transcriptionally regulate IGF1R. This study sought to identify a microRNA regulating the IGF2BPs and consequently IGF1R. All three IGF2BPs were among the top-ranked predicted targets of let-7i. Let-7i was downregulated in HCC tissues, and transfection of HuH-7 with let-7i inhibited malignant cell behaviors and decreased IGF2BPs transcripts. Direct binding of let-7i to IGF2BP2 and IGF2BP3 3'UTRs was confirmed, and the effect of let-7i caused a decrease in the IGF2BPs' target gene, the IGF1R. IGF1R mRNA was inversely correlated with let-7i in HCC tissues and was reduced upon let-7i transfection into HuH-7. Reporter assays validated IGF1R as a target of let-7i. Therefore, let-7i may control HCC tumorigenesis by regulating IGF1R directly and indirectly by interrupting the interplay between IGF1R and the IGF2BPs.
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Carcinoma Hepatocelular/metabolismo , Neoplasias Hepáticas/metabolismo , MicroRNAs/genética , Proteínas de Ligação a RNA/metabolismo , Receptores de Somatomedina/metabolismo , Regiões 3' não Traduzidas , Adulto , Carcinoma Hepatocelular/patologia , Linhagem Celular Tumoral , Feminino , Regulação Neoplásica da Expressão Gênica , Humanos , Neoplasias Hepáticas/patologia , Masculino , MicroRNAs/metabolismo , Pessoa de Meia-Idade , Proteínas de Ligação a RNA/genética , Receptor IGF Tipo 1 , Receptores de Somatomedina/genéticaRESUMO
To study the feasibility of endovascular management of early hepatic artery thrombosis (HAT) after living-donor liver transplantation (LDLT) and to clarify its role as a less invasive alternative to open surgery. A retrospective review of 360 recipients who underwent LDLT. Early HAT developed in 13 cases (3.6%). Diagnosis was performed using Doppler, CT angiography, and digital subtraction angiography. Intra-arterial thrombolysis (IAT) was performed using streptokinase or tPA. In case of underlying stricture, PTA was attempted. If the artery did not recanalize, continuous infusion was performed and monitored using Doppler US. Initial surgical revascularization was successful in 2/13 cases. IAT was performed in 11/13 cases. The initial success rate was 81.8% (9/11), the failure rate was 18.2% (2/11). Rebound thrombosis developed in 33.3% (3/9). Hemorrhage developed after IAT in 2/11 cases (18.2%). Definite endovascular treatment of HAT was achieved in 6/11 cases (54.5%) and definite treatment (surgical, endovascular or combined) in 9/13 cases (69%). (Follow-up 4 months-4 years). Endovascular management of early HAT after LDLT is a feasible and reliable alternative to open surgery. It plays a role as a less invasive approach with definite endovascular treatment rate of 54.5%.
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Procedimentos Endovasculares/métodos , Artéria Hepática/cirurgia , Transplante de Fígado/efeitos adversos , Trombose/terapia , Adulto , Angioplastia , Artéria Hepática/diagnóstico por imagem , Humanos , Doadores Vivos , Masculino , Pessoa de Meia-Idade , Estudos Retrospectivos , Terapia Trombolítica , Resultado do Tratamento , Ultrassonografia DopplerRESUMO
AIM: We have previously characterized oncogenic properties of IGF2BP1 in HCC, and its regulation by short noncoding RNAs (ncRNAs). Recent evidence suggests that IGF2BP1 itself may regulate long ncRNAs (lncRNAs). Therefore, this study aimed at exploring the interplay between IGF2BP1 and various upstream and downstream ncRNAs and its link to HCC pathogenesis. MATERIALS AND METHODS: Bioinformatic analysis was used to identify up- and downstream ncRNAs interacting with IGF2BP1. Huh-7 cells were transfected with siRNAs against IGF2BP1 and microRNA mimics. Relative gene expression was determined using RTqPCR and IGF2BP1 protein was quantified by western blot. Luciferase binding assay was used to explore the targeting of IGF2BP1 3'UTR. HCC tumorigenesis was measured by MTT assay, BrdU-incorporation assay, colony-forming assay, and scratch assay. KEY FINDINGS: Bioinformatic analysis identified three oncogenic lncRNAs - namely H19, FOXD2-AS1, and SNHG3 - potentially regulated by IGF2BP1. Knockdown of IGF2BP1 decreased the expression of all three oncogenic lncRNAs and inhibited malignant cell behaviors. miR-186 was revealed as a possible upstream regulator of IGF2BP1. miR-186 mimics decreased IGF2BP1 mRNA and protein levels. miR-186 was significantly lower while IGF2BP1 was elevated in cancerous tissues from ten HCC patients compared to five healthy controls. In addition, miR-186 mimics caused a downregulation of the oncogenic lncRNAs H19, SNHG3, and FOXD2-AS1 and a concomitant decrease in cell viability, proliferation, migration, and clonogenicity. SIGNIFICANCE: miR-186 may exert tumor suppressor effects in HCC by repressing oncogenic lncRNAs H19, SNHG3, and FOXD2-AS1 through its effect on IGF2BP1.
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Carcinoma Hepatocelular , Neoplasias Hepáticas , MicroRNAs , RNA Longo não Codificante , Proteínas de Ligação a RNA , Humanos , Carcinogênese/genética , Carcinoma Hepatocelular/patologia , Linhagem Celular Tumoral , Proliferação de Células/genética , Regulação Neoplásica da Expressão Gênica , Neoplasias Hepáticas/patologia , MicroRNAs/genética , MicroRNAs/metabolismo , RNA Longo não Codificante/genética , RNA Longo não Codificante/metabolismo , Proteínas de Ligação a RNA/genética , Proteínas de Ligação a RNA/metabolismoRESUMO
Schistosoma mansoni infection (SMI) is suspected to be directly and indirectly involved in hepato-carcinogenesis. This study evaluated the association of a previous SMI with hepatocellular carcinoma (HCC) development, patients, tumor characteristics, treatment outcomes, and survival. This observational study included patients with HCC with and without previous SMI who presented to the multidisciplinary HCC clinic, Kasr-Alainy hospital (November 2009 to December 2019). It also included 313 patients with liver cirrhosis without HCC. Clinical and laboratory features of the patients (complete blood count, liver/renal functions , alpha-fetoprotein, and hepatitis B/C status), tumor characteristics (Triphasic CT and/or dynamic MRI), liver stiffness (transient elastography), HCC treatment outcome, and overall survival were studied. This study included 1446 patients with HCC; 688(47.6%) composed group-1, defined by patients having a history of SMI, and 758(52.4%) were in group-2 and without history of SMI. Male sex, smoking, diabetes mellitus, splenomegaly, deteriorated performance status, synthetic liver functions, and platelet count were significantly higher in group-1. The groups did not differ with regard to liver stiffness, tumor characteristics, or the occurrence of post-HCC treatment hepatic decompensation or recurrence. HCC treatment response was better in group-2. Group-1 showed lower sustained virological response to hepatitis C direct-acting antivirals (DAAs) compared with group-2 (60% versus 84.3%, respectively, P = 0.027). Prior SMI was associated with HCC (adjusted odds ratio = 1.589, 95% confidence interval = 1.187-2.127), and it was concluded that it increases the risk of HCC. In addition, it significantly affects the performance status, laboratory characteristics, response to DAAs, and overall survival.
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Carcinoma Hepatocelular , Hepatite C Crônica , Neoplasias Hepáticas , Esquistossomose mansoni , Antivirais/uso terapêutico , Carcinoma Hepatocelular/tratamento farmacológico , Carcinoma Hepatocelular/patologia , Hepatite C Crônica/complicações , Humanos , Neoplasias Hepáticas/tratamento farmacológico , Neoplasias Hepáticas/patologia , Masculino , Esquistossomose mansoni/complicações , Esquistossomose mansoni/tratamento farmacológico , Esquistossomose mansoni/epidemiologiaRESUMO
BACKGROUND: Solid organ transplantation (SOT) service has been disrupted during the current coronavirus disease 2019 (COVID-19) pandemic, which deferred the service in most centers worldwide. As the pandemic persists, there will be an urgency to identify the best and safest practices for resuming activities as areas re-open. Resuming activity is a difficult issue, in particular, the decision of reopening after a period of slowing down or complete cessation of activities. OBJECTIVES: To share our experience in resuming living donor liver transplantation (LDLT) in the context of the COVID-19 pandemic in the Liver Transplantation Unit of El-Manial Specialized Hospital, Cairo University, Egypt, and to review the obstacles that we have faced. MATERIAL AND METHODS: This study is a single-center study. We resumed LDLT by the 26th of August 2020 after a period of closure from the 1st of March 2020. We have taken a lot of steps in order to prevent COVID-19 transmission among transplant patients and healthcare workers (HCWs). RESULTS: In our study, we reported three LDLT recipients, once resuming the transplantation till now. All our recipients and donors tested negative for SARS-CoV-2 by nasopharyngeal RT-PCR a day before the transplantation. Unfortunately, one of them developed COVID-19 infection. We managed rapidly to isolate him in a single room, restricting one team of HCWs to deal with him with strict personal protective measures. Finally, the patient improved and was discharged in a good condition. The second patient ran a smooth course apart from FK neurotoxicity which improved with proper management. The third patient experienced a sharp rise in bilirubin and transaminases on day 14 that was attributed to drug toxicity vs. rejection and managed by discontinuing the offending drugs and pulse steroids. In addition, one of our head nurses tested positive for SARS-CoV-2 that was manageable with self-isolation. CONCLUSION: Careful patient, donor, personnel screening is mandatory. Adequate supply of personal protective equipments, effective infection control policies, and appropriate administrative modifications are needed for a safe return of LDLT practice.
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BACKGROUND: Anti-programmed death therapy has thrust immunotherapy into the spotlight. However, such therapy has a modest response in hepatocellular carcinoma (HCC). Epigenetic immunomodulation is a suggestive combinatorial therapy with immune checkpoint blockade. Non-coding ribonucleic acid (ncRNA) driven regulation is a major mechanism of epigenetic modulation. Given the wide range of ncRNAs that co-opt in programmed cell-death protein 1 (PD-1)/programmed death ligand 1 (PD-L1) regulation, and based on the literature, we hypothesized that miR-155-5p, miR-194-5p and long non-coding RNAs (lncRNAs) X-inactive specific transcript (XIST) and MALAT-1 are involved in a regulatory upstream pathway for PD-1/PD-L1. Recently, nutraceutical therapeutics in cancers have received increasing attention. Thus, it is interesting to study the impact of oleuropein on the respective study key players. AIM: To explore potential upstream regulatory ncRNAs for the immune checkpoint PD-1/PD-L1. METHODS: Bioinformatics tools including microrna.org and lnCeDB software were adopted to detect targeting of miR-155-5p, miR-194-5p and lncRNAs XIST and MALAT-1 to PD-L1 mRNA, respectively. In addition, Diana tool was used to predict targeting of both aforementioned miRNAs to lncRNAs XIST and MALAT-1. HCC and normal tissue samples were collected for scanning of PD-L1, XIST and MALAT-1 expression. To study the interaction among miR-155-5p, miR-194-5p, lncRNAs XIST and MALAT-1, as well as PD-L1 mRNA, a series of transfections of the Huh-7 cell line was carried out. RESULTS: Bioinformatics software predicted that miR-155-5p and miR-194-5p can target PD-L1, MALAT-1 and XIST. MALAT-1 and XIST were predicted to target PD-L1 mRNA. PD-L1 and XIST were significantly upregulated in 23 HCC biopsies compared to healthy controls; however, MALAT-1 was barely detected. MiR-194 induced expression elevated the expression of PD-L1, XIST and MALAT-1. However, overexpression of miR-155-5p induced the upregulation of PD-L1 and XIST, while it had a negative impact on MALAT-1 expression. Knockdown of XIST did have an impact on PD-L1 expression; however, following knockdown of the negative regulator of X-inactive specific transcript (TSIX), PD-L1 expression was elevated, and abolished MALAT-1 activity. Upon co-transfection of miR-194-5p with siMALAT-1, PD-L1 expression was elevated. Co-transfection of miR-194-5p with siXIST did not have an impact on PD-L1 expression. Upon co-transfection of miR-194 with siTSIX, PD-L1 expression was upregulated. Interestingly, the same PD-L1 expression pattern was observed following miR-155-5p co-transfections. Oleuropein treatment of Huh-7 cells reduced the expression profile of PD-L1, XIST, and miR-155-5p, upregulated the expression of miR-194-5p and had no significant impact on the MALAT-1 expression profile. CONCLUSION: This study reported a novel finding revealing that opposing acting miRNAs in HCC, have the same impact on PD-1/PD-L1 immune checkpoint by sharing a common signaling pathway.
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BACKGROUND We assessed the alterations in portal hemodynamics associated with donor right hepatectomy and its effects on functional regeneration of the remnant liver. MATERIAL AND METHODS This prospective study included 30 adult living donors who underwent right hepatectomy in the Liver Transplantation Unit, Faculty of Medicine, Cairo University from June 2015 to October 2016. During donor surgery, portal venous pressure (PVP) was measured using an antithrombotic catheter inserted into the main portal vein, and was measured before and after clamping of the right portal vein. Postoperatively, liver function tests were done daily until normalization. The outcome measures were the time to normalization of liver function tests and effect of residual volume and steatosis on PVP. RESULTS All donors had normal PVP before clamping and changed significantly after clamping (p<0.001). After clamping, 25 donors (83%) had a PVP above 12 mmHg; i.e. had high portal pressure. The median percentage of change was 55%. There were obvious increases in liver enzymes and bilirubin after surgery, but albumin and international normalized ratio showed progressive decreases postoperatively. The percent change in PVP was positively correlated with the levels of liver enzymes, time to normalization of liver enzymes, albumin, and bilirubin, and with the degree of steatosis, bit it was negatively correlated with residual liver volume. CONCLUSIONS During living donor liver transplantation, PVP increases by over 50% after clamping of the right portal vein of the donor's liver. This increase is associated with temporary delay of normalization of liver function of the donors.
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Transplante de Fígado/métodos , Fígado/fisiologia , Doadores Vivos , Pressão na Veia Porta/fisiologia , Veia Porta/fisiologia , Adulto , Feminino , Hepatectomia , Humanos , Fígado/irrigação sanguínea , Testes de Função Hepática , Masculino , Estudos Prospectivos , Adulto JovemRESUMO
BACKGROUND: Several studies have defined the optimal portal pressure suitable for adequate graft renewal in liver transplantation (LT) but none have studied an Egyptian population to our knowledge. OBJECTIVES: Determine the level of portal venous pressure (PVP) for adequate graft function, and study the effect of PVP modulation on the outcome of LT in an Egyptian population. DESIGN: Cross-sectional, prospectively collected data. SETTING: Liver transplantation unit. PATIENTS AND METHODS: The study included adult cirrhotic pa.tients who underwent right lobe liver donor living transplantation (LDLT) at our transplantation center. Intraoperative Doppler was performed on all LDLT patients. Two PVP measurements were obtained during the recipient operation: before PV clamping and after graft re-perfusion. These PVP measurements were correlated with the results of intraoperative and postoperative Doppler findings and graft function. Mortality in the early postoperative period ( less than 1 month) and development of small-for-size syndrome (SFSS) were recorded. MAIN OUTCOME MEASURES: PVP, graft injury, and the effect of PVP modulation on the outcome of LT were the primary outcome measures. Secondary outcome measures were to correlate PVP to portal vein hemodynamics and intraoperative mean hepatic artery, peak systolic velocity, and also to correlate PVP with the postoperative graft function and mean postoperative platelet count. SAMPLE SIZE AND CHARACTERISTICS: 69 adult patients with end-stage liver disease. RESULTS: Post-reperfusion PVP was lower than pre-clamping PVP. The mean pre-clamping and post-reperfusion values were higher in patients who experienced early mortality and in patients with smaller grafts. A PVP greater than 16.5 mm Hg at the end of the operation predicted the development of SFSS (sensitivity=91.7% and specificity=50.5%). Cases of high PVP that were modulated to a lower level had a smooth and uneventful postoperative outcome. CONCLUSION: PVP is a significant hemodynamic factor that influences the functional status of the transplanted liver, including the development of SFSS, in the Egyptian population. PVP modulation may improve the outcome of LDLT. LIMITATIONS: Further study with a larger sample is needed to confirm these results. CONFLICT OF INTEREST: None.
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Transplante de Fígado/métodos , Doadores Vivos , Pressão na Veia Porta/fisiologia , Veia Porta/fisiologia , Adulto , Estudos Transversais , Egito , Feminino , Hemodinâmica , Humanos , Fígado/cirurgia , Cirrose Hepática/cirurgia , Transplante de Fígado/mortalidade , Masculino , Pessoa de Meia-Idade , Período Pós-Operatório , Estudos Prospectivos , Sensibilidade e Especificidade , Resultado do Tratamento , Ultrassonografia DopplerRESUMO
miR-let-7a is a tumor suppressor miRNA with reduced expression in most cancers. Methylation of MIRLET7A3 gene was reported to be the cause of this suppression in several cancers; however, it was not explicitly investigated in hepatocellular carcinoma (HCC). We aimed at investigating miR-let-7a expression and molecular mode in HCC, identifying drug-targetable networks, which might be affected by its abundance. Our results illustrated a significant repression of miR-let-7a, which correlated with hypermethylation of its gene of origin MIRLRT7A3. This was further supported by the induction of miR-let-7a expression upon treatment of HCC cells with a DNA-methyltransferase inhibitor. Using a computational approach, insulin-like growth factor (IGF)-II and IGF-2 mRNA binding proteins (IGF2BP)-2/-3 were identified as potential targets for miR-let-7a that was further confirmed experimentally. Indeed, miR-let-7a mimics diminished IGF-II as well as IGF2BP-2/-3 expression. Direct binding of miR-let-7a to each respective transcript was confirmed using a luciferase reporter assay. In conclusion, this study suggests that DNA hypermethylation leads to epigenetic repression of miR-let-7a in HCC cells, which induces the oncogenic IGF-signaling pathway.
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OBJECTIVES: Liver transplant performed for hepatocellular carcinoma must adhere to criteria for the size and number of focal hepatic lesions to lower the incidence of recurrence and achieve survival rates comparable to patients transplanted for other indications. Since the Milan criteria were established in 1996, there have been many less restrictive criteria yielding similar results. Our aim was to identify the prognostic factors for patient survival and for recurrence of hepatocellular carcinoma for patients within and beyond the Milan criteria. MATERIALS AND METHODS: This retrospective and prospective analysis was conducted in 60 adult patients who underwent right lobe living-donor liver transplant for cirrhosis complicated by hepatocellular carcinoma at Dar Al Fouad Hospital, 6th of October City, Egypt, between August 2001 and June 2012. The median follow-up was 39.5 months. RESULTS: Overall 1-, 3-, and 5-year survival rates were 98.3%, 93.5%, and 71.4%. Overall disease-free survival rates at 1, 3, and 5 years were 96.6%, 93.5%, and 64.2%. There was no statistically significant difference in overall survival time between patients within and beyond the Milan criteria. Factors affecting recurrence were the tumor grade, lobar distribution, size of the largest nodule, and the total tumor burden in the explanted liver. Recurrence adversely affected survival. CONCLUSIONS: Using our criteria of a single tumor ≤ 6 cm, or 2 to 3 tumors with the largest ≤ 4.5 cm, or 4 to 5 tumors with the largest ≤ 3 cm and total tumor size ≤ 8 cm resulted in overall survival comparable to patients within the Milan criteria.
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Carcinoma Hepatocelular/cirurgia , Neoplasias Hepáticas/cirurgia , Transplante de Fígado/métodos , Doadores Vivos , Adulto , Idoso , Carcinoma Hepatocelular/mortalidade , Carcinoma Hepatocelular/patologia , Intervalo Livre de Doença , Egito , Feminino , Humanos , Imunossupressores/uso terapêutico , Estimativa de Kaplan-Meier , Neoplasias Hepáticas/mortalidade , Neoplasias Hepáticas/patologia , Transplante de Fígado/efeitos adversos , Transplante de Fígado/mortalidade , Masculino , Pessoa de Meia-Idade , Gradação de Tumores , Recidiva Local de Neoplasia , Estudos Prospectivos , Estudos Retrospectivos , Fatores de Risco , Fatores de Tempo , Resultado do Tratamento , Carga TumoralRESUMO
Endothelial cell protein C receptor (EPCR) enhances the generation of activated protein C by the thrombin-thrombomodulin complex. A soluble form of EPCR (sEPCR) is present in plasma. Two polymorphisms in the EPCR gene (6936A/G and 4678G/C) have been reported to influence the risk of venous thromboembolism. We aimed to investigate the relation between EPCR gene polymorphisms (6936A/G and 4678C/G) and deep venous thrombosis (DVT) and their relations to sEPCR level. This study involved 90 patients with DVT and 90 age and sex-matched healthy controls. Plasma levels of sEPCR were measured in 45 cases of the primary DVT by ELISA. PCR-restriction fragment length polymorphism (RFLP) was used for detection of EPCR polymorphisms (6936A/G and 4678G/C). Regarding 6936A/G, our results demonstrated that mutant genotypes (AG, GG) were associated with an increased risk for DVT [Pâ<â0.001, odds ratio (OR) 4.125, 95% confidence interval (95% CI) 2.198-7.740] as well as its mutant allele G (Pâ<â0.001, OR 2.549, 95% CI 1.601-4.061). The mutant genotypes were associated with increased levels of sEPCR. Although in 4678G/C, our results demonstrated that the mutant genotype (CC) was considered as a protective factor against DVT (Pâ=â0.014, OR 0.289, 95% CI 0.108-0.776) as well as its mutant allele C (Pâ=â0.02, OR 0.600, 95% CI 0.388-0.927), but it had no effect on sEPCR level. Our data suggest that 6936A/G polymorphism is a risk factor for DVT and is associated with elevated plasma levels of sEPCR, while 4678G/C polymorphism plays a role in protection against DVT.
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Antígenos CD/genética , Polimorfismo de Nucleotídeo Único , Receptores de Superfície Celular/genética , Trombose Venosa/genética , Adolescente , Adulto , Idoso , Antígenos CD/sangue , Receptor de Proteína C Endotelial , Feminino , Genótipo , Humanos , Masculino , Pessoa de Meia-Idade , Fatores de Proteção , Receptores de Superfície Celular/sangue , Fatores de Risco , Trombose Venosa/sangue , Trombose Venosa/etiologia , Adulto JovemRESUMO
The insulin-like growth factor (IGF)-axis has been paradigmatically involved in hepatocellular carcinoma (HCC) tumor initiation, progression and drug resistance. Consequently, members of the IGF-axis and most importantly, IGF-1 receptor (IGF-1R) have been considered as intriguing targets for HCC therapy. Few miRNAs have been recently reported to be associated with IGF-1R regulation. The present study aimed to investigate the role of microRNA (miRNA/miR)-486-5p in the regulation of IGF-1R and its downstream signaling cascades. miR-486-5p was markedly downregulated in hepatitis C virus-induced HCC tissues and Huh-7 cells. Forcing the expression of miR-486-5p in Huh-7 cells resulted in the repression of IGF-1R, mammalian target of rapamycin (mTOR), signal transducer and activator of transcription 3 (STAT3) and c-Myc mRNA levels. Ectopic expression of miR-486-5p in Huh-7 cells markedly repressed cellular viability, proliferation, migration and clonogenicity in a similar pattern to IGF-1R small interfering RNAs, and were evaluated using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide, BrdU incorporation, wound healing and colony forming assays, respectively. Overall, the study findings demonstrated that miR-486-5p acts as a tumor suppressor in HCC through the repression of essential members of the IGF-axis, including IGF-1R and its downstream mediators mTOR, STAT3 and c-Myc.
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Background and Aims: The role of miR-34a in hepatocellular carcinoma (HCC) is controversial and several unresolved issues remain, including its expression pattern and relevance to tumor etiology, tumor stage and prognosis, and finally, its impact on apoptosis. Methods: miR-34a expression was assessed in hepatitis C virus (HCV)-induced non-metastatic HCC tissues by RT-Q-PCR. Huh-7 cells were transfected with miR-34a mimics and the impact of miR-34a was examined on 84 pro-apoptotic/anti-apoptotic genes using PCR array; its net effect was tested on cell viability via MTT assay. Results: miR-34a expression was up-regulated in HCC tissues. Moreover, miR-34a induced a large set of pro-apoptotic/anti-apoptotic genes, with a net result of triggering apoptosis and repressing cell viability. Conclusions: HCC-related differential expression of miR-34a could be etiology-based or stage-specific, and low expression of miR-34a may predict poor prognosis. This study's findings also emphasize the role of miR-34a in apoptosis.
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AIM: To investigate the effect of microRNA on insulin-like growth factor binding protein-3 (IGFBP-3) and hence on insulin-like growth factor-II (IGF-II) bioavailability in hepatocellular carcinoma (HCC). METHODS: Bioinformatic analysis was performed using microrna.org, DIANA lab and Segal lab softwares. Total RNA was extracted from 23 HCC and 10 healthy liver tissues using mirVana miRNA Isolation Kit. microRNA-17-5p (miR-17-5p) expression was mimicked and antagonized in HuH-7 cell lines using HiPerFect Transfection Reagent, then total RNA was extracted using Biozol reagent then reverse transcribed into cDNA followed by quantification of miR-17-5p and IGFBP-3 expression using TaqMan real-time quantitative PCR. Luciferase reporter assay was performed to validate the binding of miR-17-5p to the 3'UTR of IGFBP-3. Free IGF-II protein was measured in transfected HuH-7 cells using IGF-II ELISA kit. RESULTS: Bioinformatic analysis revealed IGFBP-3 as a potential target for miR-17-5p. Screening of miR-17-5p and IGFBP-3 revealed a moderate negative correlation in HCC patients, where miR-17-5p was extensively underexpressed in HCC tissues (P = 0.0012), while IGFBP-3 showed significant upregulation in the same set of patients (P = 0.0041) compared to healthy donors. Forcing miR-17-5p expression in HuH-7 cell lines showed a significant downregulation of IGFBP-3 mRNA expression (P = 0.0267) and a significant increase in free IGF-II protein (P = 0.0339) compared to mock untransfected cells using unpaired t-test. Luciferase assay validated IGFBP-3 as a direct target of miR-17-5p; luciferase activity was inhibited by 27.5% in cells co-transfected with miR-17-5p mimics and the construct harboring the wild-type binding region 2 of IGFBP-3 compared to cells transfected with this construct alone (P = 0.0474). CONCLUSION: These data suggest that regulating IGF-II bioavailability and hence HCC progression can be achieved through targeting IGFBP-3 via manipulating the expression of miRNAs.
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OBJECTIVES: Vascular problems such as thrombosis and stenosis of the hepatic artery, portal vein, and hepatic vein are serious complications after living-donor liver transplant and can cause increased morbidity, graft loss, and patient death. The aim of this study was to assess the incidence, treatment, and outcome of recipient vascular complications after living-donor liver transplant in a single Egyptian center. MATERIALS AND METHODS: Between November 2006 and March 2014, we performed 226 living-donor liver transplants for 225 patients at Dar Al Fouad Hospital in 6th of October City in Egypt. Review of all patients with vascular complications was performed. RESULTS: In 20 of 225 recipients (8.9%), there were vascular complications that occurred from day 0 to 14 (mean, 5.6 ± 3.4 d). Complications included isolated hepatic artery thrombosis in 7 patients (35%), isolated portal vein thrombosis in 6 patients (30%), isolated hepatic vein stenosis in 3 patients (15%), and isolated hepatic artery stenosis in 1 patient (5%). Combined portal vein thrombosis and hepatic artery thrombosis occurred in 2 patients (10%), and combined portal vein thrombosis and hepatic vein stenosis occurred in 1 patient (5%). Complications were identified with duplex ultrasonography and confirmed with computed tomographic angiography and direct angiography when needed. Multidisciplinary treatment included percutaneous transarterial or transvenous thrombolysis with or without balloon dilation and stenting, open surgical exploration with thrombectomy, vascular revision, or retransplant. There were no intraoperative deaths, but mortality occurred in 15 of 20 patients (75%). Survival ranged from 6 days to 70 months. Preoperative portal vein thrombosis was observed in 3 of 7 patients (43%) who had postoperative portal vein thrombosis. CONCLUSIONS: Major vascular complications in living-donor liver transplant recipients have poor outcome despite early detection and prompt multidisciplinary intervention. Preoperative recipient portal vein thrombosis is a risk factor for postoperative portal vein thrombosis.
Assuntos
Transplante de Fígado/efeitos adversos , Doadores Vivos , Doenças Vasculares/etiologia , Adulto , Idoso , Terapia Combinada , Diagnóstico Precoce , Egito/epidemiologia , Feminino , Humanos , Incidência , Transplante de Fígado/métodos , Transplante de Fígado/mortalidade , Masculino , Pessoa de Meia-Idade , Equipe de Assistência ao Paciente , Valor Preditivo dos Testes , Estudos Retrospectivos , Fatores de Risco , Fatores de Tempo , Tomografia Computadorizada por Raios X , Resultado do Tratamento , Ultrassonografia Doppler Dupla , Doenças Vasculares/diagnóstico , Doenças Vasculares/mortalidade , Doenças Vasculares/terapiaRESUMO
OBJECTIVES: In middle hepatic vein dominant livers, the anterior segment of the right lobe of the liver (segments V and VIII) drains mainly into the middle hepatic vein. In these donors, when right lobe grafts are procured without the middle hepatic vein, the graft may harbor large segment V and/or VIII veins that need reconstruction to avoid graft congestion and subsequent graft dysfunction. Draining these middle hepatic vein tributaries using autologous or cryopreserved vessels is a solution, despite the possible difficulties of their preparation. However, these vessels are not always available. Our objective was to evaluate the effectiveness and safety of using a synthetic vascular graft. MATERIALS AND METHODS: Between January 2012 and October 2013, eighteen adult recipients underwent living-donor liver transplant using right lobe grafts without the middle hepatic vein at Dar Al Fouad Hospital, 6th of October City, Egypt. All grafts had a large tributary of the middle hepatic vein. Eight-mm ringed expanded polytetrafluoroethylene vascular grafts were used to drain 15 segment V vein tributaries and 3 segment VIII vein tributaries directly to the inferior vena cava. Follow-up was done using duplex ultrasound to evaluate the patency of the vascular graft and the liver congestion and the liver function tests including liver enzymes. RESULTS: Intraoperative Duplex ultrasound confirmed patency and absent segmental congestion in all 18 recipients. The vascular graft patency was 17/18 at 1 week (94.4%) and 15/18 at 1 month (83.3%). No recipients developed graft infection at 1 month. CONCLUSIONS: Synthetic vascular expanded polytetrafluoroethylene grafts could be used effectively and safely in middle hepatic vein tributary reconstruction to overcome the unavailability of autologous or cryopreserved vessel grafts or just to avoid the additional burden of recovering autologous grafts thus simplifying the procedure.
Assuntos
Implante de Prótese Vascular , Veias Hepáticas/cirurgia , Transplante de Fígado/métodos , Doadores Vivos , Procedimentos de Cirurgia Plástica , Adulto , Prótese Vascular , Implante de Prótese Vascular/efeitos adversos , Implante de Prótese Vascular/instrumentação , Egito , Feminino , Veias Hepáticas/diagnóstico por imagem , Veias Hepáticas/fisiopatologia , Humanos , Testes de Função Hepática , Transplante de Fígado/efeitos adversos , Masculino , Pessoa de Meia-Idade , Politetrafluoretileno , Desenho de Prótese , Procedimentos de Cirurgia Plástica/efeitos adversos , Procedimentos de Cirurgia Plástica/instrumentação , Fatores de Tempo , Resultado do Tratamento , Ultrassonografia Doppler Dupla , Grau de Desobstrução VascularRESUMO
MicroRNAs (miRs) have a major role in the pathogenesis of hepatocellular carcinoma (HCC). As the insulin-like growth factor (IGF) axis is a highly tumorigenic pathway in HCC, the present study attempted to target it with miRs. Potential targeting of crucial members of the IGF axis by miRNAs at the 3'-untranslated region (3'-UTR) was predicted using bioinformatic tools, such as microrna.org, Diana lab and Targetscan, while 5'-UTR targeting was predicted using bibiserv software. Expression profiling of obtained miRNAs was performed using quantitative polymerase chain reaction (qPCR) in 22 non-metastatic HCC biopsy samples and 10 healthy tissues. To investigate the impact of miRNAs on their potential downstream targets, transfection of miRNAs was performed in HuH-7 cells and the targets' expression was quantified using qPCR. Transcripts of insulin-like growth factor-1 receptor (IGF-1R), insulin-like growth factor binding protein-3 (IGFBP-3) and IGF-II were found to be potentially targeted at the 5'-UTR and 3'-UTR regions by the single clustered hepatic metastamiRs miR-96-5p and miR-182-5p. The two miRNAs showed a similar expression pattern in HCC tissues compared to those in healthy tissues. Forced expression of miR-96-5p and miR-182-5p in the HCC cell line HuH-7 had inducing effects on IGFBP-3 and IGF-II transcripts. Of note, the two miRs had differential effects on IGF-1R, where miR-96-5p induced IGF-1R mRNA expression and miR-182-5p inhibited its expression. The present study revealed the pleiotropic impact of the single clustered hepatic metastamiRs miR-96-5p and miR-182-5p on IGF-1R, and an inducing effect on IGF-II and IGFBP-3 in hepatocellular carcinoma.
Assuntos
Carcinoma Hepatocelular/genética , Proteína 3 de Ligação a Fator de Crescimento Semelhante à Insulina/biossíntese , Fator de Crescimento Insulin-Like II/biossíntese , Neoplasias Hepáticas/genética , MicroRNAs/genética , Receptor IGF Tipo 1/biossíntese , Carcinoma Hepatocelular/patologia , Linhagem Celular Tumoral , Regulação Neoplásica da Expressão Gênica , Humanos , Proteína 3 de Ligação a Fator de Crescimento Semelhante à Insulina/genética , Fator de Crescimento Insulin-Like II/genética , Neoplasias Hepáticas/patologia , Receptor IGF Tipo 1/genética , TransfecçãoRESUMO
Hepatocellular carcinoma (HCC) is characterized by the aberrant expression of a number of genes that govern crucial signaling pathways. The insulin-like growth factor (IGF) axis is important in this context, and the precise regulation of expression of members of this axis is known to be lost in HCC. miR-155 is a well-established oncogene in numerous types of cancer. However, to the best of our knowledge, its effect on the regulation of the IGF axis has not been investigated to date. The present study aimed to elucidate the interactions between miR-155 and key components of the IGF axis, in addition to examining its effect on HCC development. Reverse transcription-quantitative polymerase chain reaction was used to measure the expression of miR-155 in HCC and cirrhotic tissues, in addition to HCC cell lines. Furthermore, the effect of the induction of miR-155 expression on the expression of three members of the IGF axis [IGF II, IGF type-1 receptor (IGF-1R) and IGF-binding protein 3 (IGFBP-3)], was analyzed. Finally, the effect of miR-155 on HCC cell proliferation, migration and clonogenicity was also examined. Quantification of the expression of miR-155 demonstrated that it is upregulated in HCC. Induction of the expression of miR-155 in HCC cell lines led to the upregulation of IGF-II and IGF-IR, and the downregulation of IGFBP-3. In addition, the proliferation, migration and clonogenicity of HCC was increased following induction of miR-155 expression. miR-155 is an oncomiR, which upregulates the oncogenes, IGF-II and IGF-IR, and downregulates the tumor suppressor, IGFBP-3, thereby resulting in increased HCC cell carcinogenicity. Therefore, miR-155 may be a therapeutic target in HCC.
RESUMO
This study aimed to identify a single miRNA or miR (microRNA) which regulates the three insulin-like growth factor-2-mRNA-binding proteins (IGF2BP1, 2 and 3). Bioinformatics predicted miR-1275 to simultaneously target the three IGF2BPs, and screening revealed miR-1275 to be underexpressed in hepatocellular carcinoma (HCC) tissues. Transfection of HuH-7 cells with miR-1275 suppressed IGF2BPs expression and all three IGF2BPs were confirmed as targets of miR-1275. Ectopic expression of miR-1275 and knockdown of IGF2BPs inhibited malignant cell behaviors, and also reduced IGF1R protein and mRNA. Finally IGF1R was validated as a direct target of miR-1275. These findings indicate that the tumor-suppressor miR-1275 can control HCC tumor growth partially through simultaneously regulating the oncogenic IGF2BPs and IGF1R.