Targeting TRMT5 suppresses hepatocellular carcinoma progression via inhibiting the HIF-1α pathways / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

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.

Methyl rosmarinate induces cell apoptosis in human hepatocellular carcinoma cells via inhibiting the Akt/mTOR signaling pathways / 西安交通大学学报(医学版)

【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.

Analysis of the therapeutic efficacy of transcatheter arterial chemoembolization com-bined with systemic treatment in unresectable hepatocellular carcinoma / 中国肿瘤临床

Objective:To investigate the efficacy and safety of different transcatheter arterial chemoembolization(TACE)-based regimens in patients with unresectable hepatocellular carcinoma(uHCC)and explore the optimal timing for combining TACE with tyrosine kinase inhibit-ors(TKIs)and immune checkpoint inhibitors(ICIs).Methods:A retrospective analysis was conducted on data from 555 patients with uHCC who underwent TACE-based treatment between April 2016 and December 2021 in Nanfang Hospital,Southern Medical University.The pa-tients were assigned into the following four groups according to different treatment regimens:TACE group(n=317),TACE combined with TKIs group(TACE+TKIs,n=66),TACE combined with ICIs group(TACE+ICIs,n=33),and TACE combined with TKIs+ICIs group(TACE+TKIs+ICIs,n=139).Subgroup analysis was performed within the TACE+TKIs+ICIs group,with patients being assigned into"pre-TACE"and"post-TACE"groups based on the timing of the combination therapy.Univariate and multivariate Cox regression analyses were conducted to identify pro-gnostic factors influencing overall survival(OS).Results:The TACE+TKIs+ICIs group showed the longest OS(21.9 months,95%confidence in-terval[CI]:17.2-26.6,P=0.030)and progression-free survival(PFS)(8.3 months,95%CI:7.3-9.3,P=0.004)compared to those in the other three groups.In the subgroup analysis,the"post-TACE"group had longer OS than the"pre-TACE"group(26.8 months vs.19.2 months,P = 0.011).The objective response rate(ORR)was 32.8%,41.1%,42.4%,and 52.5%(P=0.001)and the disease control rate(DCR)was 59.6%,71.2%,69.7%,and 82.7%(P<0.001)in the TACE,TACE+TKIs,TACE+ICIs,and TACE+TKIs+ICIs groups,respectively.The adverse events were similar to those reported in previous studies.Cox regression analysis revealed that tumor number,extrahepatic metastasis,and treatment regimen were independent factors influencing OS in patients(all P<0.05).Conclusions:TKIs or ICIs can improve OS and PFS in patients with uHCC receiving TACE,and the combination of TKIs+ICIs with TACE achieves better beneficial outcomes.The greatest OS was observed when the combination therapy TKIs+ICIs was initiated within 3 months after the first TACE procedure.

Mechanisms of Euphorbia fischeriana steud. alcohol extracts intreatment of HCC based on network pharmacology and experiment verification / 中国药理学通报

Aim To investigate the molecular mechanisms of alcohol extracts of Euphorbia fischeriana steud. against hepatocellular carcinoma (HCC) through a combination of network pharmacology analysis and experimental validation. Methods The active ingredients and targets of alcohol extracts of Euphorbia fischeriana steud. were determined through TCMSP, Swiss ADME, Swiss Target Prediction database and references. The databases DisGeNET and GeneCards were employed to screen potential HCC-related genes. Venny platform, STRING platform and Cytoscape software were applied to construct active ingredient-target-disease and protein-protein interaction (PPI) network maps. Gene ontology (GO) and kyoto encyclopedia of genes and genomes (KEGG) enrichment analyses were performed using the DAVID database. To assess the effects of Euphorbia fischeriana steud. alcohol extracts on BEL-7402 cells, the proliferation and apoptosis were detected by CCK-8, EdU and flow cytometry assays, and the related protein levels of JAK2/STAT3 pathway were analyzed by Western blot. Additionally, H22 hepatocellular carcinoma mouse model was used to evaluate the in vivo efficacy of Euphorbia fischeriana steud. alcohol extracts. Results A total of 916 HCC targeted genes, 30 active ingredients containing the related 567 potential targeted genes, and 115 intersection targets of disease and compounds were obtained. KEGG enrichment analysis identified JAK2/STAT3 signaling as a critical pathway. In vitro experiments showed the alcohol extracts of Euphorbia fischeriana steud. could inhibit proliferation, promote apoptosis and suppress JAK2/STAT3 signaling pathway in a dose-dependent manner in BEL-7402 cells. In addition, the alcohol extracts of Euphorbia fischeriana steud., either alone or in combination with sorafenib, dramatically blocked tumor growth in in vivo tests. Conclusions Euphorbia fischeriana steud. alcohol extracts have anti-cancer effects in HCC, and the molecular mechanisms may be connected to the regulation of JAK2/STAT3 signaling pathway.

Mechanism of Transcription Factor SP1 Regulates Expression of Tripartite Motif-containing Protein 7 in Hepatocellular Carcinoma / 中国生物化学与分子生物学报

Tripartite motif containing protein 7 (TRIM7), as a member of the E3 ubiquitin ligase TRIM family, plays an important regulatory role in immune regulation, metabolism and other physiological processes. The aberrant expression of TRIM7 is closely related to the development and progression of hepatocellular carcinoma (HCC) and it shows a complex regulatory role. However, the regulatory mechanism for the expression of TRIM7 in HCC remains unknown. In this study, multiple online databases were used to analyze the expression of TRIM7 in HCC and data indicated that TRIM7 expression was upregulated in HCC and correlated to poor prognosis. Subsequently, the transcription factor binding sites in the TRIM7 promoter region were analyzed using UCSC and JASPAR databases, and the results showed that TRIM7 promoter contains four SP1 binding sites. In this work, we demonstrated that SP1 could directly bind to its binding sites in TRIM7 promoter and positively regulate the transcriptional activity driven by the TRIM7 promoter using dual luciferase reporter experiments and the ChIP-PCR method. Moreover, our results also showed SP1 overexpression upregulated the expression of TRIM7 at both mRNA and protein levels (P<0. 01),and SP1 inhibitor, mithramycin A, could reverse the activated effect of SP1 on TRIM7 expression (P<0. 01). In conclusion, this study preliminarily reveals the regulatory mechanism of TRIM7 upregulation in HCC, which provides an important theoretical basis for further study of the gene function, early diagnosis and targeted therapy.

Advances in Molecular Targeted Drugs for Hepatocellular Carcinoma / 中山大学学报(医学科学版)

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.

Clinical Study of Jiawei Xiaochaihutang Combined with Microwave Ablation in Treatment of Primary Hepatocellular Carcinoma / 中国实验方剂学杂志

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.

Deubiquitinase JOSD2 Promotes the Survival and Metastasis of Hepatoma Cells / 中国生物化学与分子生物学报

Ubiquitination is a reversible post-translational modification that regulates protein stability and plays a function in numerous signaling pathways. Deubiquitinases play important roles in the regulation of the occurrence and development of different cancers. In this study, we analyzed the survival and prognosis of liver cancer patients according to the expression levels of 63 deubiquitinases. Results discovered that JOSD2 expression in tumor was significantly higher than that in the adjacent tissue (P<0. 0001) and was negatively associated with overall survival of patients (P < 0. 05). JOSD2 is a deubiquitinase from MJD sub-family. Other members of this sub-family were not correlated with liver cancer. The pathways related to cell proliferation were significantly enriched (FDR < 0. 05) in a differential gene function enrichment analysis of JOSD2 high-expressing samples in TCGA data. In hepatoma cell lines, we demonstrated that the overexpression of JOSD2 significantly enhances cell survival, migration and invasion. In conclusion, this study found that JOSD2 is highly expressed in liver cancer (P = 0. 041), and that patients with high-expressed JOSD2 have significantly shorter overall survival. Furthermore, overexpression of JOSD2 can promote the survival and metastasis of hepatoma cells (P < 0. 01), which suggests that JOSD2 might promote the survival and metastasis of hepatocellular carcinoma.

The Expression and Functional Roles of Epoxide Hydrolase 2 in Hepatocellular Carcinoma / 中国生物化学与分子生物学报

The expression, function and prognostic significance of epoxide hydrolase 2 (EPHX2) in hepatocellular carcinoma (HCC) were comprehensively analyzed through collecting HCC tissues and public database. The GEO and MitoCarta databases were used to identify the mitochondria-related differentially expressed genes (DEGs) in HCC. The Cancer Genome Atlas (TCGA) database was applied to analyze the expression levels of DEGs in HCC, including EPHX2 and its co-expressed genes. The R package was applied to draw the Kaplan-Meier survival curve and gene function enrichment analysis. The STRING database and GSEA software were used to analyze the protein-protein interaction (PPI) network and gene set enrichment analysis. qPCR and GEO database were applied to verify the expression level of EPHX2 in HCC. In the present study, a total of 15 mitochondria-related DEGs were identified in HCC. The expression of EPHX2 in HCC was significantly decreased compared to the normal liver tissues (P < 0. 01). The expression of EPHX2 was related to gender, tumor stage and grade in HCC, but not associated with age, T stage, et al in HCC. Moreover, compared with the patients with lower expression of EPHX2, patients with higher expression of EPHX2 had a better prognosis. EPHX2 was associated with fatty acid degradation. In addition, PPI results indicated that HAO1, AGXT, ACOX1, GSTκ1, SCP-2, CAT, CYP2C8, CYP2C9, CYP2B6, and CYP2J2 were co-expressed with EPXH2 in HCC. Furthermore, GSEA results showed that the group with lower expression of EPHX2 was positively correlated with the gene set of liver cancer cell proliferation and liver cancer recurrence. qPCR and GEO database results verified that the expression of EPHX2 was significantly decreased in HCC. The expression of EPHX2 was decreased in HCC, strongly suggesting that EPHX2 might function as a tumor suppressor gene in HCC. However, the potential mechanism of EPHX2 in HCC needs to be further verified.

TRIM15 Knockdown Induces Cell Cycle Arrest in HepatocellularCarcinoma Cells / 中国生物化学与分子生物学报

Tripartite motif-containing protein 15 (TRIM15) is a member of the TRIM family, which is a class of proteins with E3 ubiquitin ligase activity. The function of TRIM15 in tumors is rarely reported. This study is intended to explain the role of TRIM15 in hepatocellular carcinoma. Nuclear and cytoplasmic fractionation and immunofluorescence assays confirmed that TRIM15 was located in the nucleus and cytoplasm. We designed hairpin RNA (shRNA) to knockdown TRIM15 in hepatocarcinoma cell lines. After knocking down TRIM15, cell growth curve and clone formation assays showed that cell proliferation was significantly inhibited (P<0. 05). Cell cycle analysis by flow cytometry showed that knockdown of TRIM15 blocked cell cycle in the G

SC⁃BPSO: A Filter Fused BPSO Feature Selection Method in Hepatocellular Carcinoma Classification / 中国生物化学与分子生物学报

Early diagnosis of cancer can significantly improve the survival rate of cancer patients, especially in patients with hepatocellular carcinoma (HCC). Machine learning is an effective tool in cancer classification. How to select high⁃classification accuracy feature subsets with low dimension in complex and high⁃dimensional cancer datasets is a difficult problem in cancer classification. In this paper, we propose a novel feature selection method, SC⁃BPSO: a two⁃stage feature selection method implemented by combining the Spearman correlation coefficient, chi⁃square independent test⁃based filter method, and binary particle swarm optimal (BPSO) based wrapper method. It has been applied to the cancer classification of high⁃dimensional data to classify normal samples and HCC samples. The dataset in this paper is obtained from 130 liver tissue microRNA sequence data (64 hepatocellular carcinoma, 66 normal liver tissue) from National Center for Bioinformatics (NCBI) and European Bioinformatics Institute (EBI). First, the liver tissue microRNA sequence data was preprocessed to extract the three types of features of microRNA expression, editing level and post⁃editing expression. Then, the parameters of the SC⁃BPSO algorithm in the liver cancer classification were adjusted to select a subset of key features. Finally, classifiers were used to establish classification models, predict the results, and compare the classification results with the feature subset selected by the information gain filter, the information gain ratio filter and the BPSO wrapper feature selection algorithm using the same classifier. Using the feature subset selected by the SC⁃BPSO algorithm, the classification accuracy is up to 98. 4%. The experimental results showed that compared with the other three feature selection algorithms, the SC⁃ BPSO algorithm can effectively find feature subsets with relatively small size and higher accuracy. This may have important implications for cancer classification with a small number of samples and high⁃ dimension features.

THBS4 Promotes the Metastasis of Hepatocellular Carcinoma Cells by Inducing M2 Polarization of Tumor-associated Macrophages / 中国生物化学与分子生物学报

Thrombospondin 4 (THBS4), a member of the THBS family, is a protein secreted by the extracellular matrix and is involved in regulating various physiological processes, such as cell proliferation, adhesion and angiogenesis. Recent studies have shown that the inflammation stimulates THBS4 production and induces the adhesion and accumulation of macrophages. Our previous study confirmed that THBS4 acts as an oncogene in hepatocellular carcinoma (HCC), the effect of THBS4 on the immune microenvironment of HCC remains unclear. This study aims to analyze the role of THBS4 in promoting the metastasis of HCC cells by inducing M2-type polarization of tumor-associated macrophages. We simulate the tumor microenvironment through HCC conditioned medium (HCM) and found that the expression of THBS4 in macrophages increased in a time-dependent manner under the action of HCM (P<0.05); THBS4 knockdown promotes the expression of M1 macrophages markers IL-1β and CD86 (P<0.01), while the expression of M2-type markers IL-10 and CD206 were decreased (P<0.01). Transwell co-culture assay was used to further detect the effect of THBS4-induced M2-type macrophages on HCC metastasis. Results from co-culture of THBS4-downregulated M2 macrophages with HepG2 cells showed that THBS4-downregulated M2-TAMs significantly inhibited the invasion and migration ability of HepG2 cells (all P < 0.01). In conclusion, the tumor microenvironment promotes the expression of THBS4 in macrophages, and THBS4 may promote the invasion and metastasis of HCC cells by inducing M2-type polarization of macrophages. This study provides some new experimental basis for exploring the establishment of THBS4-induced HCC immune microenvironment.

Key Genes of Peripheral Blood Mononuclear Cells in Hepatocellular Carcinoma and Effective Chinese Herbs： A Bioinformatics Analysis and Systematic Review / 中国实验方剂学杂志

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⁺and CD4⁺/CD8⁺levels， and reducing the serum CD8⁺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.

Immune response triggered by the ablation of hepatocellular carcinoma with nanosecond pulsed electric field / 医学前沿

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.

Expression of HHLA2 in hepatocellular carcinoma tissues and its clinical significance / 中国肿瘤生物治疗杂志

@#[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.

CAAP1 inhibits apoptosis and promotes proliferation, migration and invasion of hepatocellular carcinoma HepG2 cells / 中国肿瘤生物治疗杂志

@#[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.

Value of preoperative magnetic resonance imaging features in predicting early recurrence of hepatocellular carcinoma after surgical resection / 中国肿瘤临床

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.

Summary of the research frontiers of liver transplantation in 2020 ATC / 器官移植

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.

Upregulation of OGT by Caveolin-1 Promotes Mouse Hepatocellular Carcinoma Cells Migration and Invasion / 中国生物化学与分子生物学报

Caveolin-1 (Cav-1), a major structural protein of caveolae, is implicated in the vesicular uptake processes of transcytosis and cell signaling. However, its role in modulating protein glycosylation and tumor metastasis remains to be further elucidated. In the present study, it was shown that Cav-1 promotes the expression of O-GlcNAcylation and O-GlcNAc transferase (OGT), and triggers the invasion and metastasis of hepatocellular carcinoma (HCC) cells. The results of RT-qPCR, Western blot and dual lucif-erase reporter assay showed that Cav-1 negatively regulated the expression of transcription factor RUNX2 in HCC. Subsequently, this results in attenuate RUNX2-induced transcription of miR24. miR24 suppresses mouse HCC cells invasion and metastasis via directly targeting Ogt mRNA 3′UTR. This research provides evidence of Cav-1-mediated OGT expression and O-GlcNAc (O-linked N-acetylglucosamine) elevation. These data give insight into a novel mechanism of HCC occurrence and development.

Prospect of Clinical Application of Glypican-3 Targeted Therapy for Hepatocellular Carcinoma / 中国生物化学与分子生物学报

Glypican-3 (GPC3) is an oncofetal protein anchored on the plasma membrane and highly expressed in hepatocellular carcinoma (HCC).GPC3 could he used as a biomarker for the diagnosis of HCC and the serum levels of GPC3 in liver cancer patients has a significant role for their prognosis.Moreover, GPC3 in HCC cells is immunoreactive, rendering it a suitable target for the treatment of HCC.Nowadays, several clinical trials targeting GPC3 for HCC therapy have already been conducted: new anti- GPC3 antibodies are generated; the clinical trials about its combination administration with other targeted medicines are being in progress; (tP(3-targeted TRAB, GPC3 vaccines and GPC3-based chimeric antigen receptor T-cells (CAR-T) therapy are under investigation.In this review, we briefly discuss the structure of GPC3, its role in HCC pathogenesis and summarize the recent development in the clinical application of GPC3.We firmly believe that GPC3 would be a promising target for HCC therapy.And further studies focusing on GPC3 should provide us more solid evidence.