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Accumulating evidence has confirmed the links between transfer RNA (tRNA) modifications and tumor progression. The present study is the first to explore the role of tRNA methyltransferase 5 (TRMT5), which catalyzes the m1G37 modification of mitochondrial tRNAs in hepatocellular carcinoma (HCC) progression. Here, based on bioinformatics and clinical analyses, we identified that TRMT5 expression was upregulated in HCC, which correlated with poor prognosis. Silencing TRMT5 attenuated HCC proliferation and metastasis both in vivo and in vitro, which may be partially explained by declined extracellular acidification rate (ECAR) and oxygen consumption rate (OCR). Mechanistically, we discovered that knockdown of TRMT5 inactivated the hypoxia-inducible factor-1 (HIF-1) signaling pathway by preventing HIF-1α stability through the enhancement of cellular oxygen content. Moreover, our data indicated that inhibition of TRMT5 sensitized HCC to doxorubicin by adjusting HIF-1α. In conclusion, our study revealed that targeting TRMT5 could inhibit HCC progression and increase the susceptibility of tumor cells to chemotherapy drugs. Thus, TRMT5 might be a carcinogenesis candidate gene that could serve as a potential target for HCC therapy.
Subject(s)
Humans , Carcinoma, Hepatocellular/pathology , Cell Hypoxia , Cell Line, Tumor , Gene Expression Regulation, Neoplastic , Hypoxia-Inducible Factor 1, alpha Subunit/metabolism , Liver Neoplasms/pathology , Signal Transduction/genetics , tRNA Methyltransferases/metabolismABSTRACT
【Objective】 To investigate the cell death-inducing effect of methyl rosmarinate (MR) on human hepatoma Hep-3B and SK-Hep1 cells and their potential mechanisms. 【Methods】 The effects of MR on the viability of Hep-3B, SK-Hep1 and MIHA cells were determined by cell counting kit-8 (CCK-8) assay. The morphological changes of three kinds of cells treated with different concentrations of MR were observed by optical microscopy. EdU assay and flow cytometry were used to detect the proliferation and apoptosis of Hep-3B and SK-Hep1 cells. Transwell assay was used to study the effects of MR on the migration and invasion of Hep-3B and SK-Hep1 cells. Western blotting was used to evaluate the protein expression levels of apoptosis, EMT and Akt/mTOR signaling pathways. 【Results】 After treated with different concentrations of MR (0~200 μmol/L) for 48 h, Hep-3B and SK-Hep1 cells activities were significantly decreased in a concentration-dependent manner (P<0.01), while there was no significant effect on MIHA cell activity (P>0.05), and the IC50 of Hep-3B and SK-Hep1 cells were 102.5 and 99.3 μmol/L, respectively. MR treatment (0-150 μmol/L) significantly inhibited the proliferation of Hep-3B and SK-Hep1 cells (P<0.05), while cell detachment and shrinkage were observed by optical microscopy on the Hep-3B and SK-Hep1 cells, while the morphology of MIHA cells was not changed. Compared with the control group, MR (100, 150 μmol/L) induced apoptosis in Hep-3B and SK-Hep1 cells, the expression levels of the pro-apoptotic proteins Bax and cleaved PARP were significantly increased (P<0.05), while the expression level of the anti-apoptotic protein Bcl-2 was significantly decreased (P<0.05). MR (100, 150 μmol/L) also inhibited the migration and invasion of HCC cells, significantly increased the expression of E-cadherin and decreased the expression of N-cadherin and Vimentin compared with the control group (P<0.05). Finally, Western blotting results showed that the expressions of p-Akt and p-mTOR in Hep-3B and SK-Hep1 treated by MR were significantly reduced in a dose-dependent manner, suggesting that MR may play an anti-cancer role by inhibiting Akt/mTOR signaling pathway. 【Conclusion】 MR can promote apoptosis in Hep-3B and SK-Hep1 cells, which may be closely related to the inhibition of Akt/mTOR signaling pathway.
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Being the most common solid malignant tumor in the digestive system and the third leading cause of cancer-related death worldwide， hepatocellular carcinoma （HCC） is characterized by insidious onset， early recurrence/metastasis and poor prognosis. With the advantages of targeted precision， high specificity， minimal drug resistance， remarkable therapeutic efficacy and fewer side effects， molecular targeted drugs have become the hotspot and focus of tumor therapy research in recent years. As more is learned about the mechanism and clinical efficacy， some molecular targeted drugs have been recommended by HCC treatment guidelines. This paper reviewed the mechanism of HCC targeted therapy， molecular targeted drugs， relevant therapeutic protocols and outcomes so as to provide reference and evidence for subsequent research.
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ObjectiveTo observe the clinical efficacy of Jiawei Xiaochaihutang combined with microwave ablation （MWA） in the treatment of primary hepatocellular carcinoma （HCC） and its influence on tumor microenvironment. MethodA total of 128 patients were randomly divided into control group （64 cases： 2 cases of dropout，2 cases of elimination，and 60 cases of completion） and observation group （64 cases： 3 cases of dropout，2 cases of elimination，and 59 cases of completion）. Both groups were given comprehensive treatment after MWA surgery. Patients in control group took Biejiajian Wan orally （3 g/time，3 times/d）， and those in observation group took Jiawei Xiaochaihutang （1 dose/d）. The treatment lasted for 3 consecutive months. The size of solid tumor before and after treatment was evaluated to record the progression-free survival （PFS）. The alpha-fetoprotein-L13 （AFP-L3），des-γ-carboxy prothrombin （DCP），Golgi protein 73 （GP73），tumor necrosis factor-α （TNF-α），transforming growth factor-β （TGF-β），vascular endothelial growth factor （VEGF） and matrix metalloproteinase-2 （MMP-2） levels，as well as performance status （PS），liver function and syndrome of liver depression and Qi stagnation scores were also detected before and after treatment. In addition， the incidence of side effects of grade Ⅲ and above was compared. ResultThe total effective rate of solid tumor in observation group was 91.53% （54/59），higher than that （76.67%， 46/60） in control group（χ2=4.895，P<0.05）. The PFS in observation group was （7.16±0.95） months， longer than that （6.24±0.89 months） in control group （P<0.01）. The effective rate of traditional Chinese medicine （TCM） syndrome in observation and control groups were 88.14% （52/59）and 70.00% （42/60）， respectively （χ2=5.897，P<0.05）. The observation group （57.63%，34/59） had higher marked effective rate of TCM syndrome than control group （31.67%，19/60） （χ2=8.116，P<0.01）. The AFP-13，DCP，GP73，TNF-α，TGF-β，VEGF and MMP-2 levels and the PS，liver function and syndrome of liver depression and Qi stagnation scores in observation group were lower than those in control group （both P<0.01）. The cumulative incidence of side effects of grade Ⅲ and above in observation and control groups was 16.95% and 33.33%， respectively（χ2=4.261，P<0.05）. ConclusionConsolidation treatment of HCC after MWA surgery with Jiawei Xiaochaihutang relieved symptoms and side effects，improved PS and liver function，regulated tumor microenvironment，inhibited tumor markers and prolonged survival time. The clinical effect was better than that of Biejia decoction pill， and thus it was worthy of clinical use.
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The American Transplant Congress (ATC) is an influential academic congress in the field of organ transplantation. In this article, the hotspots of liver transplantation in 2020 ATC were summarized, including the latest research progress in donor liver procurement and quality assessment, donor liver preservation and ischemia-reperfusion injury (IRI), liver transplantation for hepatocellular carcinoma and other hepatic malignancies, complications after liver transplantation, transplantation immunology, perioperative management and donor-derived infection, pediatric liver transplantation and cell therapy, etc.
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Objective: To assess the value of preoperative clinical data and magnetic resonance (MR) imaging features in predicting early recurrence (recurrence in two years) after surgical resection of hepatocellular carcinoma (HCC). Methods: This retrospective study included 244 patients with HCC who underwent a surgical resection between January 2015 and January 2018 at Tianjin Medical University Cancer Institute and Hospital. The role of preoperative clinical data and MR imaging features on early recurrence after surgical tumor resection were evaluated using univariate and multivariate analyses. All patients were followed up regularly after discharge. The endpoint was considered to be intrahepatic recurrence within 2 years. Results: In the univariate analyses, the maximum diameter of the tumor, tumor capsule, peritumoral parenchyma enhancement, rim enhancement, two-trait predictor of venous invasion (TTPVI), tumor necrosis, satellite nodules, dynamic enhancement pattern, diffusion-weighted imaging (DWI) /T2WI mismatch and other MR imaging features, as well as alpha-fetoprotein (AFP), TNM stage, alanine aminotransferase (ALT), glutamatergic aminotransferase (AST), direct bilirubin (DBIL), γ-glutamyl transferase (γ-GT) and other clinical data were correlated with the early recurrence of HCC. In the multivariate Cox regression analysis, the tumor capsule (HR=0.372, P400 μg/L (HR=2.234, P400 μg/L were found to be independent factors of the early postoperative recurrence of HCC. This research has established a predictive model for the early recurrence of HCC after surgical resection using a non-invasive method, which can help clinicians to develop individualized treatment protocols and improve patient outcomes.
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@#[Abstract] Objective: To explore the effect of CAAP1 on apoptosis, proliferation, migration and invasion of hepatocellular carcinoma (HCC) HepG2 cells and its mechanism. Methods: The pcDNA3/CAAP1 (CAAP1 over-expression) and pSilencer 2.1-U6 neo/shR-CAAP1 (CAAP1 knockdown) plasmids were constructed and transfected into HepG2 cells. The mRNA and protein levels of CAAP1 were detected by qPCR and WB, respectively. The cells were divided into four groups, namely overexpression control group (pcDNA3), CAAP1 over-expression group (pcDNA3/CAAP1), silence control group (pSilencer 2.1-U6 neo, pSilencer) and CAAP1 silence group (pSilencer 2.1-U6 neo/shR-CAAP1, shR-CAAP1). Flow cytometry was used to analyze the apoptosis, and WB was used to detect the protein expression of cleaved caspase 3 in each group. CCK-8 assay was used to detect the proliferation of HepG2 cells, Colony formation assay was used to detect the clonogenesis, and Transwell assay and wound healing assay were used to detect the invasion and migration abilities of HepG2 cells in each group. The effect of CAAP1 on overall survival (OS) of HCC patients was analyzed after searching TCGA database. Results: PcDNA3/CAAP1 with CAAP1 over-expression and shR-CAAP1 with CAAP1 knockdown were successfully constructed. It was confirmed that pcDNA3/CAAP1 could increase the mRNA and protein expressions of CAAP1, while shR-CAAP1 could decrease the mRNA and protein expressions of CAAP1 (all P<0.05). The cell apoptotic rate in pcDNA3/CAAP1 group decreased by 32% as compared to pcDNA3 group, and the cleaved caspase 3 protein expression was significantly decreased (all P<0.05); while the cell apoptotic rate in shR-CAAP1 group increased by 73% as compared to pSilencer group, and the cleaved caspase 3 protein expression was significantly increased (all P<0.05). The cell proliferation in pcDNA3/CAAP1 group significantly increased (P<0.05), while the cell proliferation in shR-CAAP1 group significantly decreased (P<0.05). The cell migration number increased by 48%, the cell migration distance increased by 59% (P<0.05) and the cell invasion number increased by 52% in pcDNA3/CAAP1 group (all P<0.05). The cell migration number decreased by 53%, cell migration distance decreased by 29% and cell invasion number decreased by 45% in shR-CAAP1 group (all P<0.05). TCGA database analysis showed that the high expression of CAAP1 was negatively correlated with the OS of HCC patients (P<0.05). Conclusion: CAAP1 can promote the proliferation, migration and invasion of HepG2 cells by inhibiting its apoptosis, and it may be closely related to the occurrence and development of HCC.
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@#[Abstract] Objective: To investigate the expression of human endogenous retrovirus subfamily H long terminal repeat associating protein 2 (HHLA2) in hepatocellular carcinoma (HCC) tissues and its correlation with the clinicopathological characteristics and prognosis of patients with HCC. Methods: Based on TCGA database, the correlation between HHLA2 mRNA expression and B7 family genes in human HCC tissues was analyzed. HHLA2 expression in 90 pairs of HCC tissues and their adjacent tissues was detected by tissue microarry and immunohistochemical staining. Wilcoxon rank sum test was used to compare the difference of HHLA2 expression between HCC tissues and its adjacent tissues. The chi-square test was used to analyze the relationship between HHLA2 expression in human HCC tissues and clinicopathological features of the patients. Kaplan-Meier survival analysis was performed to analyze the correlation between HHLA2 expression and patients’ overall survival (OS), and the Cox model was used to evaluate the prognostic value of different indices. Results: The expression level of HHLA2 mRNA in HCC tissues was correlated with B7 family CD274, C10orf54, PDCD1LG2, ICOSLG and CD276. The expression level of HHLA2 in HCC tissues was significantly correlated with tumor size (χ2=4.531, P<0.05). The OS of HCC patients with high HHLA2 expression was significantly shorter than that of the patients with lower HHLA2 expression (HR=1.878, 95%CI: 1.066-3.309, P<0.05). The COX model showed that tumor size (HR=2.493, 95%CI: 1.310-4.742, P<0.01) could be used as an independent risk factor for the prognostic prediction of the patients. Conclusion: HHLA2 is significantly correlated with the prognosis of HCC patients, and can be used as a potential target for HCC immunotherapy.
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Objective:To explore the key gens of peripheral blood mononuclear cells in hepatocellular carcinoma（HCC-PBMC） and potentially effective Chinese herbs based on bioinformatics， and to verify the clinical efficacy of these Chinese herbs via a systematic review. Method:The chips GSE58208 and GSE36076 of HCC-PBMC were downloaded from the Gene Expression Omnibus （GEO）， followed by the identification of differentially expressed genes （DEGs） using RStudio. After protein-protein interaction （PPI） analysis by STRING， the DAVID was employed for gene ontology （GO） and Kyoto Encyclopedia of Genes and Genomes （KEGG） enrichment analysis. The DEGs of HCC-PBMC were visualized by Cytoscape. The key genes of HCC-PBMC were calculated by CytoHubba plug-in and mapped with those in Coremine Medical for screening out the potential Chinese herbs for the treatment of HCC， which were then included for subsequent systematic review. Result:A total of 203 DEGs were obtained （194 up-regulated and nine down-regulated）. As revealed by DAVID analysis， the DEGs were mainly enriched in such biological processes and signaling pathways as transcriptional regulation of DNA template， hydrolysis of ribonucleic acid phosphodiester bond， positive regulation of intranuclear mitosis and division， skeletal muscle fiber development， activation of mitogen-activated protein kinase（MAPK）activity， Fanconi anemia pathway， and metabolic pathway. The key genes of HCC-PBMC were calculated by Cytoscape to be<italic> </italic>GTPase IMAP family member 1 （GIMAP1）， GTPase IMAP family member 4 （GIMAP4）， GTPase IMAP family member 6 （GIMAP6）， GTPase IMAP family member 7 （GIMAP7）， GTPase IMAP family member 8 （GIMAP8）， interleukin-1<italic>β </italic>（IL-1<italic>β）</italic>， CX3C chemokine receptor 1 （CX3CR1）， C-C chemokine receptor type 2 （CCR2）， Toll-like receptor 7（TLR7）， and epidermal growth factor（EGF）. Through Coremine Medical analysis， it was concluded that Ginseng Radix et Rhizoma， Curcumae Longae Rhizoma， Centellae Herba， and Hedyotidis Herba were closely related to the key genes. Ginseng Radix et Rhizoma has the effects of tonifying and benefiting lung and spleen and enhancing strength， suitable for the liver depression and spleen deficiency syndrome or Qi-Yin deficiency syndrome of HCC. Hence， Si Junzitang with Ginseng Radix et Rhizoma as the sovereign medicinal was included for systematic review. It has been confirmed that Ginseng Radix Et Rhizoma was superior to western medicine alone in improving the overall clinical efficacy， alleviating TCM syndrome， elevating serum CD4<sup>+ </sup>and CD4<sup>+</sup>/CD8<sup>+ </sup>levels， and reducing the serum CD8<sup>+ </sup>and TBIL levels （<italic>P</italic><0.01）， with high safety. Conclusion:This study conducted at the gene level has provided new ideas for clinical diagnosis and treatment of HCC. The systematic review of Ginseng Radix Et Rhizoma against HCC provides a basis for the clinical prevention and treatment of HCC with TCM.
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Nanosecond pulsed electric field (nsPEF) is a novel, nonthermal, and minimally invasive modality that can ablate solid tumors by inducing apoptosis. Recent animal experiments show that nsPEF can induce the immunogenic cell death of hepatocellular carcinoma (HCC) and stimulate the host's immune response to kill residual tumor cells and decrease distant metastatic tumors. nsPEF-induced immunity is of great clinical importance because the nonthermal ablation may enhance the immune memory, which can prevent HCC recurrence and metastasis. This review summarized the most advanced research on the effect of nsPEF. The possible mechanisms of how locoregional nsPEF ablation enhances the systemic anticancer immune responses were illustrated. nsPEF stimulates the host immune system to boost stimulation and prevail suppression. Also, nsPEF increases the dendritic cell loading and inhibits the regulatory responses, thereby improving immune stimulation and limiting immunosuppression in HCC-bearing hosts. Therefore, nsPEF has excellent potential for HCC treatment.
Subject(s)
Animals , Carcinoma, Hepatocellular/therapy , Cell Line, Tumor , Immunity , Liver Neoplasms/therapy , Neoplasm Recurrence, LocalABSTRACT
Objective: Hepatic cancer is known as primary liver cancer and hepatocellular carcinoma (HCC). Newly silver nanoparticles gained importance due to its advantages and multiple potential such as molecular imaging agent, antimicrobial, wound healing, anti-inflammatory and anticancer activity. The current study deals to assess therapeutic property silver nanoparticles (AgNPs) against diethylnitrosamine (DENA), and carbon tetrachloride (CCL4) induced hepatic cancer. Methods: Thirty male albino rats (200-250g) were distributed into four groups and hepatic cancer was induced with a single intraperitoneal dose of 200 mg/kg body weight of DENA. Two weeks later, animals received subcutaneous injections of CCl4 once a week in a dose of 3 ml/kg body weight for 6weeks. Serum biomarkers, antioxidants enzymes, inflammatory markers were evaluated to find the anti-proliferative potential of silver nanoparticles. Histological evaluation and microscopic reports were also done to document the results of the current work. Results: AgNPs significantly recover the serum marker enzymes of hepatic parameter AST, ALT, ALP, and total bilirubin and also decreased the levels of NO, IL-6 and TNF-α. Histopathological features also exhibited recovery of a hepatic architecture in cancer-induced rats. Moreover, the immunohistochemical investigation demonstrated that the levels of PCNA, and Caspase-3, which are hepatocarcinogenic markers, were significantly improved by AgNPs. Conclusion: These results concluded that AgNPs showed promising curing effects on hepatocellular ailments.
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Cancer is a disease in which a group of abnormal cells grow uncontrollably by disregarding the normal rules of cell division. Across several cancers, Hepatocellular carcinoma (HCC) is one of the most aggressive cancers in worldwide. It is held responsible for up to 1 million deaths globally per annum. HCC is an inflammation-related cancer, as a chronic inflammatory state is necessary for cancer appearance. In this study, the drug astaxanthin and encapsulated astaxanthin was tested against HCC. Mice were divided into 7 groups; Group I: control, Group II: DEN induced, Group III: DEN + 50 mg/kg astaxanthin, Group IV: DEN + 100 mg/kg astaxanthin, Group V: DEN + 50 mg/kg encapsulated astaxanthin, Group VI: DEN + 100 mg/kg encapsulated astaxanthin, Group VII: DEN + 10 mg/kg sorafenib. Regular diet was given. Body weight, Food intake, water intake was noted. Other biochemical parameters such as ALP, AST, Albumin, proteins and TNF-α was determined. Finally, the liver was removed from each mice of different group by sacrificing them and histopathology was done. In vivo evaluation in mice models showed significant antitumor activities by both encapsulated and non-encapsulated astaxanthin at 100 mg/kg as compared with the control, DEN induced group and positive drug sorafenib. This research suggested that encapsulated astaxanthin can also be used as chemotherapeutic agent for the treatment of Hepatocellular carcinoma (HCC).
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Objective: To investigate the value of the Child-Pugh (CTP), ALBI, MELD, and MELD-Na scores in predicting acute-on-chronic liver failure (ACLF) in patients with hepatocellular carcinoma(HCC) after transcatheter arterial chemoembolization (TACE). Methods: Seven hundred and eleven patients with HCC who received their first TACE treatment at Guangxi Medical University Cancer Hospital between October 2013 and October 2015 were retrospectively analyzed. A Logistic regression analysis and receiver operating characteristic (ROC) curve were used to evaluate the efficacy of the four scoring models in predicting ACLF. Results: The results of the univariate and multivariate analysis showed that the four scoring models could independently predict the occurrence of ACLF after TACE. The ROC curve analysis showed that the area under the ROC curve (AUC) of ALBI was significantly higher than the other three scores (P5.5, ALBI >-2.29, MELD >8.08 and MELD-Na >8.08 was higher than those with scores lower than the cut-off values (P0.001). Conclusions: The child-Pugh, ALBI, MELD, and MELD-Na scores have certain predictive value for ACLF after TACE treatment, with ALBI having the best predictive value.
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Objective: To investigate the expression level, clinical significance and mechanisms of miR-3127-5p in hepatocellular carcinoma (HCC). Methods:Real-time quantitative PCR (qPCR) was performed to detect miR-3127-5p expression in HCC tissues and the adjacent non-tumor tissues as well as in HCC cell lines and normal hepatic cells. The difference in miR-3127-5p expresssion between normal liver tissues and HCC tissues was analyzed by TCGA database. The relationship of miR-3127-5p expression with the clinicopathologic features and prognosis of HCC patients was analyzed. After the HCCLM3 cells were transfected with miR-3127-5p inhibitors or negative control sequence, the abilities of cell migration and invasion were measured by Transwell assay; the potential downstream of miR-3127-5p was predicted by TargetScan and validated by Luciferase reporter assay. Results: Compared with that in tumor adjacent tissues or normal hepatic cells, miR-3127-5p was increased in HCC tissue and cell lines (all P<0.05). The expression of miR-3127-5p was significantly related to the portal vein tumor thrombus (P=0.016) and advanced TNM stage (P=0.016). The result of overall survival investigated by TCGA database showed that the survival rate in patients with high miR-3127-5p expression was significantly lower than that in those with low miR-3127-5p expression (P=0.0023). Knockdown of miR-3127-5p significantly inhibited HCCLM3 cell migration and invasion compared with negative controls. Cyclin-dependent kinase 10 (Cdk10), the potential downstream of miR-3127-5p, was validated by luciferase reporter assay. Rescue assay showed that Cdk10 mediated the effects of miR-3127-5p on migration and invasion of HCC cells. Conclusion: miR-3127-5p is upregulated in HCC, which promotes HCC cell migration and invasion through targeting Cdk10.
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@#Uncontrolled chronic inflammation plays key roles in the carcinogenesis of hepatocellular carcinoma (HCC). Among the risk factors of HCC, such as chronic viral hepatitis, alcoholic hepatitis, non-alcoholic steatohepatitis and so on, the occurrence and development of uncontrolled chronic inflammation are the core factors of HCC. The damaged or dead hepatocytes generated during the process of chronic inflammation may lead to the activation of immune cells in the liver, resulting in hepatic inflammation. Chronic and prolonged liver inflammation promotes the occurrence of cancer. During this process, different injuries or death patterns of hepatocytes and progression of inflammation caused by activation of different immune cells play different roles in hepatic carcinogenesis, involving multiple pathological or pathophysiological processes such as liver injury, inflammation, and compensatory proliferation, as well as function alteration of various cells, signaling pathways, and regulatory molecules. Further studies on the regulatory mechanisms of hepatic inflammation-induced carcinogenesis are helpful to provide theoretical basis for the intervention of occurrence of HCC. This review focused on the research progress of regulatory mechanisms involved in the hepatic inflammation-induced carcinogenesis.
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@#[Abstract] Objective: To explore the regulatory effect of long non-coding RNA (lncRNA) SNHG5 on invasion and migration of hypoxia-induced hepatocellular carcinoma (HCC) cells. Methods: A total of 20 pairs of cancer and para-cancerous tissue specimens resected from HCC patients in the First Affiliated Hospital of Xi'an Jiaotong University from January 2017 to June 2018, and human HCC cell lines (HepG2, MHCC-97L, MHCC-97H , Huh7) as well as immortalized human liver LO2 cells were collected for this study. Bioinformatics methods were used to analyze the binding sites between hypoxia-inducible factor 1α (HIF-1α) and SNHG5. pCMVHIF-1α and shRNA-SNHG5 (sh-SNHG5) plasmids were transfected into HCC cells, respectively. qPCR was used to detect the expres‐ sion level of SNHG5 in HCC tissues and hypoxia-induced HCC cells. Western botting was used to detect the expression level of HIF-1α protein in HCC cells, and Transwell chamber method was used to detect the migration and invasion ability of HCC cells after SNHG5 si‐ lence under normoxia and hypoxia condition. Results: Compared with para-cancerous tissues and immortalized human liver LO2 cells, the expression of SNHG5 was significantly up-regulated in HCC tissues and cell lines (all P<0.01). Hypoxia promoted the expression level of SNHG5 in HCC cells, and its mechanism might be related to the combination of hypoxia-activated HIF-1α and SNHG5 promoter to promote its transcription. Hypoxia promoted the invasion and migration ability of HepG2 and MHCC-97L cells (all P< 0.01), but knockdown of SNHG5 significantly inhibited the invasion and migration ability of HepG2 and MHCC-97L cells under hy‐ poxic conditions (all P<0.01). Conclusion: SNHG5 is highly expressed in HCC tissues and cell lines and plays an important role in the invasion and migration of HCC cells induced by hypoxia.
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@#Objective: To identify the specific Hub genes in young hepatocellular carcinoma (HCC) patients, and to explore their biological and clinical significance by using bioinformatic methods. Methods: The data information of HCC and normal tissues of young (≤40 years old at diagnosis) and old (＞40 years old at diagnosis) HCC patients were obtained from GEO chip data set GSE45267. The differentially expressed genes (DEGs) in HCC tissues as comparing to normal tissues in the two groups were screened by using GEO2R and Venn chart software. The Protein-Protein Interaction (PPI) network of the specific DEGs in young group was constructed by bioinformatics tools STRING and Cytoscape to screen the Hub genes and significant modules. The Hub genes were verified by GEPIA database, and the overall survival time was analyzed by Kaplan-Meier. Finally, Gene Ontology (GO) Enrichment analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis were used to analyze the DEGs specific to young group and the common DEGs of the two groups by DAVID. Results: Finally, 117 up-regulated and 179 down-regulated DEGs specific to the young group were screened out, and PPI network screened 10 most connected genes as Hub genes, among which 7 Hub genes were concentrated in the first module. Six up-regulated Hub genes, including TYMS, CDC6, BUB1, TPX2, OIP5 and KIF23, were indicated to associate with the poor prognosis in young HCC patients by GEPIA and Kaplan-Meier analysis. GO function and KEGG pathway analyses showed that the DEGs specific to young HCC patients were mainly involved in biological processes such as ATP binding, and were mainly enriched in S phase of cell cycle; while the common DEGs of two groups were mainly involved in biological processes such as cyclooxygenase P450 and cell division, and were mainly enriched in the G2/M phase of the cell cycle. Conclusion: In this study, 6 up-regulated DEGs specific to young group that suggested poor prognosis were identified, which may be the potential therapeutic and prognostic targets for young patients with HCC.
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@# Objective: To investigate the expression of CARD10 in hepatocellular carcinoma (HCC) tissues, and the roles of CARD10 in HCC progression especially apoptosis inhibition. Methods: The expression of CARD10 was examined in pared non-tumor liver tissues and HCC tissues using qRT-PCR, and their correlation with HCC TNM stage was analyzed using Spearman’s rank correlation assay in SPSS 17.0. In HCC cells with CARD10 overexpression or knockdown, cytometry using Annexin-V/PI labeling was used to measure apoptosis, and Western blotting was used to determine the activation of NF-κB pathway. Results: CARD10 expression was significantly increased in HCC tissues as compared to that in pared non-tumor liver tissues (P<0.01), and the increased CARD10 in HCC was positively correlated with TNM staging (P<0.01). The apoptosis of HCC cell lines SMMC-7721 and BEL-7402 was inhibited by CARD10 overexpression while promoted by CARD10 knockdown, and the pro-survival NF-κB pathway was also enhanced by CARD 10 over-expression while suppressed by CARD10 knockdown. Conclusion: CARD10 expression is increased in HCC tissues and positively correlated with HCC progression. CARD10 inhibits HCC apoptosis by promoting the activation of NF-κB pathway. [Key words] hepatocellular carcinoma (HCC); caspase recruitment domain family member 10 (CARD10); apoptosis; NF-κB
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@# Objective: To investigate the effect and mechanism of RNA binding protein Lin28 on the 5-fluorouracil (5-Fu) sensitivity of HepG2 cells. Methods: HepG2 cells were transfected with plasmid pcDNA3.1-Lin28 or si-Lin28 (small interfering RNA of Lin28). qPCR and Western blotting were used to detect the expression of Lin28 in HepG2 cells after transfection. Changes of cell proliferation in transfected cells after 5-Fu treatment was detected by CCK8 assay and the 50% inhibitory concentration (IC50) was calculated. Flow cytometry was used to detect apoptotic rate after 5-Fu treatment and the expression of apoptosis-related protein was assayed by Western blotting. The mRNA expressions of drug-resistant miRNAs (let-7a and let-7b), as well as cancer stem cell markers (Oct4, Nanog and Sox2) after transfection were detected by qPCR. Results: As compared to the HepG2/Vector cells, the mRNA and protein expressions of Lin28 were significantly up-regulated in HepG2/Lin28 cells (P<0.05 or P<0.01). Over-expression of Lin28 significantly suppressed the sensitivity of HepG2 cells to 5-Fu (IC50elevated obviously, P<0.05) and significantly increased cell proliferation while decreased apoptotic rate and expression of apoptotic-related protein caspase-3 (all P<0.01). As compared to si-control group, expression of Lin28 in HepG2/si-Lin28 cells was significantly down-regulated (P<0.01). Lin28 knockdown significantly reduced cell proliferation and IC50 of 5-Fu (all P<0.01) but increased apoptotic rate and expression of apoptosis-related protein (P<0.01). Compared with HepG2/Vector group, expressions of let-7a and let-7b, as well as cancer stem cell markers (Oct4, Nanog and Sox2) were significantly increased in HepG2/Lin28 cells (all P<0.01); while these molecules were significantly decreased in HepG2/si-Lin28 cells as comparing to si-control group (all P<0.01). Conclusion: Lin28 can modulate the chemosensitivity of HepG2 cells by regulating the expression of miRNAs and the formation of cancer stem cells. Targeting Lin28 might be a promising approach to improve the chemotherapy efficacy in HCC.
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@# Objective：To explore the targeting relationship between miR-377-5p and hypoxia inducible factor-1 (HIF-1α), and investigate the regulatory effect of miR-377-5p on proliferation, invasion and epithelial-mesenchymal transition (EMT) of hepatocellular carcinoma (HCC) cells through vascular endothelial growth factor (VEGF) signaling pathway. Methods： ：The expression of miR-377-5p in 35 pairs of human HCC tissues and para-cancerous tissues was detected by qPCR. Then, HepG2 cells were divided into control group, mimic-NC group and miR-377-5pmimicgroup.qPCRwasusedto detect the transfection efficiency; the effects of miR-377-5p over-expression on proliferation and invasion of HepG2 cells were examined by EdU staining and Transwell assay, respectively; and the effect of miR-377-5p over-expression on the expressions of proliferation-related protein Ki-67, proliferating cell nuclear antigen (PCNA) and epithelial-mesenchymal transition (EMT) markers (E-cadherin and N-cadherin) were detected by Western blotting (WB); the effect of miR-377-5p over-expression on the expression of hypoxia inducible factor-1α (HIF-1α) in HepG2 cells was detected by qPCR and WB; and the targeting relationship between miR-377-5p and HIF-1α gene was determined by Luciferase reporter gene assay. Results: The expression of miR-377-5p in HCC tissues was significantly lower than that in para-cancerous tissues (P<0.01). Compared with the control group, the expression of miR-377-5p in HepG2 cells of miR-377-5p mimic group elevated significantly, and the proliferation, invasion and the expression of N-caderin proteins decreased,significantly (all P<0.01), while the expression of E-caderin increased significantly (P<0.01). At the same time, the mRNA and protein expressions of HIF-1α in miR-377-5p mimic group decreased significantly (P<0.01 or P<0.05). miR-377-5p targetedly inhibited the expression of HIF-1α gene and suppressed the activation of VEGF pathway (all P<0.05). Conclusion: miR-377-5p inhibits the proliferation, invasion and EMT of HepG2 cells via targetedly inhibiting HIF-1α expression and suppressing the activation of VEGF signaling pathway.