Adjacent Liver Tissues Research Articles

Autocrine osteopontin is involved in maintaining the growth and metastasis of Echinococcus multilocularis

Alveolar echinococcosis is a zoonotic disease that seriously endangers human health. This study aims to investigate the effects of osteopontin on the growth and intra- or extra-hepatic metastasis of Echinococcus multilocularis. Mice were randomly divided into untreated (control group, n = 25), PBS (n = 25), Lv3-NC (n = 25), and Lv-OPN-734 (n = 25) groups. Knockdown OPN by injecting lentivirus through the intraperitoneal portal vein, the metastatic lesions infected with Echinococcus multilocularis and adjacent liver tissues were observed, and the expression of osteopontin and epidermal growth factor receptor pathway-related molecules were studied. Gross observation of specimens suggested that there was no extra- hepatic metastasis, and mild intrahepatic invasion was observed in the Lv-OPN-734 group after 4 months of infection, and lung metastasis occurred in the Lv3-NC group. Western-blot and immunohistochemical staining results showed that the protein expression of OPN, phosphorylation of epidermal growth factor receptor and downstream molecules of the pathway decreased significantly after osteopontin knockdown, whereas the levels of non-phosphorylated proteins did not change significantly. In human tissues, through western-blot and immunohistochemical staining we found that compared with the control group, the expression of OPN in the liver tissues infected with Echinococcus multilocularis were higher than that in the control group. These findings indicate that osteopontin is involved in maintaining the growth and metastasis of Echinococcus multilocularis, suggesting that osteopontin may be a potential target for the treatment of alveolar echinococcosis.

SHC4 promotes tumor proliferation and metastasis by activating STAT3 signaling in hepatocellular carcinoma

BackgroundThe Src homology and collagen 4 (SHC4) is an important intracellular adaptor protein that has been shown to play a pro-cancer role in melanoma and glioma. However, the biological function and detailed mechanisms of SHC4 in hepatocellular carcinoma progression are unclear. This study aimed to evaluate the potential prognostic and treatment value of SHC4 in patients with HCC.MethodsThe expression status of SHC4 in HCC tissues were investigated by immunohistochemistry and western blotting. Clinical significance of SHC4 was evaluated in a large cohort of HCC patients. The effects of SHC4 repression or overexpression on migration, invasion, and tumor growth were detected by colony formation assay, wound healing, transwell assays, and xenograft assay. Cell cycle and EMT-related proteins were detected by western blotting and immunofluorescence. In addition, the molecular regulation between SHC4 and STAT3 signaling in HCC were discovered by western blotting, immunofluorescence and xenograft assay.ResultsSHC4 was overexpressed in HCC compared to adjacent normal liver tissues and increased SHC4 expression was associated with high AFP level, incomplete tumor encapsulation, poor tumor differentiation and poor prognosis. SHC4 was shown to enhance cell proliferation, colony formation, cells migration and invasion in vitro, and promotes cell cycle progression and EMT process in HCC cells. Tumor xenograft model assay confirmed the oncogenic role of SHC4 in tumorigenicity in nude mice. Moreover, activation of STAT3 signaling was found in the SHC4 overexpressed HCC cells and HCC tissues. Further intervention of STAT3 confirmed STAT3 as an important signaling pathway for the oncogenic role of SHC4 in HCC.ConclusionsTogether, our results reveal that SHC4 activates STAT3 signaling to promote HCC progression, which may provide new clinical ideas for the treatment of HCC.

Gene signature to predict prognostic survival of hepatocellular carcinoma.

Hepatocellular carcinoma (HCC) has a high incidence and poor prognosis and is the second most fatal cancer, and certain HCC patients also show high heterogeneity. This study developed a prognostic model for predicting clinical outcomes of HCC. RNA and microRNA (miRNA) sequencing data of HCC were obtained from the cancer genome atlas. RNA dysregulation between HCC tumors and adjacent normal liver tissues was examined by DESeq algorithms. Survival analysis was conducted to determine the basic prognostic indicators. We identified competing endogenous RNA (ceRNA) containing 15,364 pairs of mRNA–long noncoding RNA (lncRNA). An imbalanced ceRNA network comprising 8 miRNAs, 434 mRNAs, and 81 lncRNAs was developed using hypergeometric test. Functional analysis showed that these RNAs were closely associated with biosynthesis. Notably, 53 mRNAs showed a significant prognostic correlation. The least absolute shrinkage and selection operator’s feature selection detected four characteristic genes (SAPCD2, DKC1, CHRNA5, and UROD), based on which a four-gene independent prognostic signature for HCC was constructed using Cox regression analysis. The four-gene signature could stratify samples in the training, test, and external validation sets (p <0.01). Five-year survival area under ROC curve (AUC) in the training and validation sets was greater than 0.74. The current prognostic gene model exhibited a high stability and accuracy in predicting the overall survival (OS) of HCC patients.

Fluorescence lifetime needle optical biopsy discriminates hepatocellular carcinoma.

This work presents results of in vivo and in situ measurements of hepatocellular carcinoma by a developed optical biopsy system. Here, we describe the technical details of the implementation of fluorescence lifetime and diffuse reflectance measurements by the system, equipped with an original needle optical probe, compatible with the 17.5G biopsy needle standard. The fluorescence lifetime measurements observed by the setup were verified in fresh solutions of NADH and FAD++, and then applied in a murine model for the characterisation of inoculated hepatocellular carcinoma (HCC) and adjacent liver tissue. The technique, applied in vivo and in situ and supplemented by measurements of blood oxygen saturation, made it possible to reveal statistically significant transformation in the set of measured parameters linked with the cellular pools of NADH and NADPH. In the animal model, we demonstrate that the characteristic changes in registered fluorescent parameters can be used to reliably distinguish the HCC tissue, liver tissue in the control, and the metabolically changed liver tissues of animals with the developed HCC tumour. For further transition to clinical applications, the optical biopsy system was tested during the routing procedure of the PNB in humans with suspected HCC. The comparison of the data from murine and human HCC tissues suggests that the tested animal model is generally representative in the sense of the registered fluorescence lifetime parameters, while statistically significant differences between their absolute values can still be observed.

Comprehensive kinomic study via a chemical proteomic approach reveals kinome reprogramming in hepatocellular carcinoma tissues.

Hepatocellular carcinoma (HCC) is one of the most common cancers worldwide. Kinases are attractive therapeutic targets since they are commonly altered in cancers. Here, to identify kinases of potential therapeutic interest in HCC, a quantitative kinomic study of tumour and adjacent non-tumour liver tissues was performed using a chemical proteomics approach. In total, 124 kinases were found differentially expressed and they were distributed over all nine kinase groups. Exploration of The Cancer Genome Atlas (TCGA) data showed that the dysregulation of 45 kinases was correlated with poor prognosis in HCC patients. We then tested 11 inhibitors targeting 12 crucial protein kinases alone or in combination for their ability to inhibit cell growth in Hep3B and PLC/PRF/5 cell lines. Six inhibitors significantly reduced viability in both cell lines. Combination inhibition of polo-like kinase 1 (PLK1) and casein kinase 1 epsilon (CSNK1E) significantly induced growth arrest in both cell lines synergistically. In summary, our analysis presents the most complete view of kinome reprogramming in HCC and provides novel insight into crucial kinases in HCC and potential therapeutic targets for HCC treatment. Moreover, the identification of hundreds of differentially expressed kinases forms a rich resource for novel drug targets or diagnostic biomarker discovery. Data are available via ProteomeXchange (identifier PXD023806).

Proteogenomic characterization identifies clinically relevant subgroups of intrahepatic cholangiocarcinoma.

We performed proteogenomic characterization of intrahepatic cholangiocarcinoma (iCCA) using paired tumor and adjacent liver tissues from 262 patients. Integrated proteogenomic analyses prioritized genetic aberrations and revealed hallmarks of iCCA pathogenesis. Aflatoxin signature was associated with tumor initiation, proliferation, and immune suppression. Mutation-associated signaling profiles revealed that TP53 and KRAS co-mutations may contribute to iCCA metastasis via the integrin-FAK-SRC pathway. FGFR2 fusions activated the Rho GTPase pathway and could be a potential source of neoantigens. Proteomic profiling identified four patient subgroups (S1-S4) with subgroup-specific biomarkers. These proteomic subgroups had distinct features in prognosis, genetic alterations, microenvironment dysregulation, tumor microbiota composition, and potential therapeutics. SLC16A3 and HKDC1 were further identified as potential prognostic biomarkers associated with metabolic reprogramming of iCCA cells. This study provides a valuable resource for researchers and clinicians to further identify molecular pathogenesis and therapeutic opportunities in iCCA.

Operative Management of Liver Abscess and Intra-hepatic Migration of Gallstones due to Perforated Cholecystitis

We report a rare case and an atypical presentation of acute cholangitis and cholecystitis complicated by intrahepatic gallbladder perforation, forming a multilocular liver abscess with stones embedded within the abscess cavity in a forty-six-year-old gentleman. He presented with painless jaundice, pale stools, and dark urine. His initial investigations revealed leukocytosis, obstructive liver function tests and computerised tomography of the abdomen revealed a perforated gallbladder with segment 5/6 liver abscess containing multiple dependent calculi in segment 6 and distal CBD calculi. Endoscopic-Retrograde-Cholangiopancreatography was performed, with endoscopic sphincterotomy and retrieval of common bile duct stones. This was followed by ultrasound-guided percutaneous cholecystectomy. He was then treated with la[aroscopic cholecystectomy, deroofment and debridement of abscess and retrieval of intrahepatic stones. In the setting of hepatic abscess, percutaneous drainage and antibiotic treatment for abscess before an interval cholecystectomy, however the retained calculi within the abscess cavity in this case continue to be a nidus for infection. With decompression of the abscess by drainage and regeneration of scar tissue, the difficulty of retrieving the stones requires resection of the involved liver segments. The early timing and conduct of the operation allowed the use of a minimally invasive parenchymal-sparing approach to address the intrahepatic stones along with the cholecystectomy. While perforated gallbladder with cholecystohepatic fistula and abscess is a rare condition on its own; we believe this to be the first time an abscess from a perforated gallbladder has been complicated by migration of gallstones beyond the adjacent pericholecystic liver tissues and into non-continuous segments.

Expression of cyclo-oxygenase-2 and yap/taz in hepatocellular carcinoma in untreated and treated hepatitis C virus patients.

The pathogenesis of hepatitis C virus (HCV)-related hepatocellular carcinoma (HCC) differs according to whether prior treatment with interferon (IFN) vs. direct-acting antiviral agents (DAAs) was administered. Cyclo- oxygenase-2 (COX-2), yes-associated protein 1 (YAP), and transcriptional co-activator with PDZ-binding motif (TAZ) play acrucial role in hepatocarcinogenesis. However, their roles in untreated or treated HCV-related HCC development have not been clarified. Therefore, we performed an immunohistochemical study and stained tissue from 83 HCV-related HCC cases using antibodies against COX-2, YAP, and TAZ and correlated their expression with the clinicopathological characteri stics and survival data. The cases were subdivided into 3 groups based on prior HCV treatment. In the 3 groups, COX-2 was significantly higher in HCC tissue compared with adjacent non-tumour liver tissue. However, the expression of YAP/TAZ was not significantly different between HCC and adjacent non-tumour tissue. We further grouped HCC cases into YAP+/TAZ+ and YAP-/TAZ- cases. In the YAP+/TAZ+ cases, COX-2 was significantly associated with tumour size, tumour multifocality, and late pathologic stage. No significant difference was observed in COX-2 and TAZ expression as aresult of IFN or DAA treatment; however, YAP was significantly higher in IFN-treated HCC. Cyclo-oxygenase-2 overexpression may play arole in late HCC development, while YAP/TAZ could play an early role in HCC progression. Sustained expression of combined YAP/TAZ could mediate the poor prognostic role of COX-2.

The diagnostic and prognostic significance of small nuclear ribonucleoprotein Sm D1 aberrantly high expression in hepatocellular carcinoma.

Small nuclear ribonucleoprotein Sm D1 (SNRPD1), one of the crucial genes encoding core spliceosome components, was abnormally highly expressed in multiple types of tumors. In this study, we investigated the diagnostic and prognostic significance of SNRPD1 in hepatocellular carcinoma (HCC). The investigation of datasets from GEO and TCGA databases revealed that SNRPD1 expression in HCC was significantly higher than adjacent normal liver tissues, which was validated by immunohistochemistry (IHC). Both GO, KEGG analysis showed that the SNRPD1 co-expressed genes mainly enriched in Cell division, Nuclear import, mRNA splicing via spliceosome, Ribosome, Cell cycle, etc. Survival analysis from the GSE14520 dataset and 154 HCC cohorts exhibited a significant association of high SNRPD1 expression with poor overall survival and recurrence-free survival. ROC analysis showed that the abnormally high SNRPD1 mRNA expression has diagnostic significance in distinguishing between HCC and normal liver tissue (AUC = 0.819). Gene set enrichment analysis (GSEA) demonstrated that the high expression of SNRPD1 might regulate HCC tumorigenesis and progression by affecting the cell cycle, mismatch repair, DNA replication, and RNA degradation, etc. The luciferase report assay revealed that SNRPD1 was the direct target gene of miR-100 manifested by decreased SNRPD1 expression and luciferase activity in the HCC cells upon miR-100 overexpression. Finally, SNRPD1 may as an oncogene affecting the progression of HCC through regulates the mTOR pathway and autophagy.

Overexpressed GNAZ predicts poor outcome and promotes G0/G1 cell cycle progression in hepatocellular carcinoma.

We aimed to investigate the role of G protein subunit alpha Z(GNAZ) in the progression and prognosis of patients with hepatocellular carcinoma (HCC). Oncomine, GEO, TCGA, GEPIA2, Kaplan-Meier Plotter, TIMER2, Metascape, CCLE, LinkedOmics, and UALCAN databases were used to analyze the differential expression of GNAZ in HCC and normal liver tissues, relationship between GNAZ expression and prognosis of patients with HCC, and expression of GNAZ in common human HCC cell lines. Western blotting was performed to analyze GNAZ expression, while the Cell Counting Kit 8 assay was used to determine cell proliferation, and flow cytometry was used to evaluate the cell cycle and apoptosis. Wound healing and transwell invasion assays were used to investigate cell metastasis and invasion. Using Oncomine, Gene Expression Omnibus (GEO), and GEPIA2 databases, GNAZ was found to be overexpressed in HCC tissues compared with that in adjacent normal liver tissues, and western blotting analysis showed GNAZ overexpression in seven patients with HCC who underwent surgical resection of HCC and para-cancerous tissues (p<0.01). Survival analysis revealed that high GNAZ expression was negatively associated with overall survival (OS), recurrence-free survival, progression-free survival, and disease-specific survival in patients with HCC (p<0.05). GNAZ overexpression was associated with worse 4- month, 6- month, 12- month, 24- month, 36- month, 48- month, and 60-month OS, as well as with different clinicopathological characteristics of patients with HCC, including hepatitis virus infection state; alcohol consumption state; male; female; Asian; microvascular invasion, Stage I-II, Stage II-III, and Stage III-IV; and grade II (Cox regression, p<0.05). KEGG/GO biological process enrichment indicated that the genes similar to GNAZ in HCC were mainly enriched in the cell cycle, cell cycle phase transition, DNA replication checkpoint, and regulation of G0 to G1 transition. siRNA-GNAZ significantly reduced the viability of JHH-2 and SNU-761 cells from 12 to 96h; increased the percentage of cells in the G0/G1 phase and decreased that of cells in the S and G2/M phases (p<0.05); and markedly downregulated the expression of cyclin D, cyclin E, and CDK2 protein. siRNA-GNAZ also significantly increased the percentage of JHH-2 and SNU-761 cell apoptosis at late stages, while the number of surviving cells decreased (p<0.05), and upregulated the expression of apoptosis-related proteins Bax and caspase 3 protein. Furthermore, siRNA-GNAZ remarkably reduced the healing of scratch wounds in JHH-2 and SNU-761 cells and the number of invasive cells compared with that in the control group (p<0.001). Our study demonstrated that GNAZ plays a pivotal role as a potential oncogene and predicts poor prognosis in patients with HCC. It promotes tumor proliferation via cell cycle arrest, apoptosis, migration, and invasion. Thus, GNAZ may be a potential candidate biomarker providing useful insight into hepatocarcinogenesis and aggressiveness.

PHF21A expression as a biomarker of hepatocellular carcinoma progression and prognosis.

Previous studies have shown that PHF21A is associated with the initiation and progression of various tumors. However, its role in hepatocellular carcinoma (HCC) is still unclear. Thus, this study aimed to determine the expression and clinical significance of PHF21A in HCC. PHF21A expression in 201 liver cancer samples and 129 adjacent normal tissues was detected by immunohistochemistry. The correlation between PHF21A expression and the clinicopathological features and prognosis of HCC was verified in 70 other liver tissue microarray samples. The relationship between PHF21A expression and HCC immune cell infiltration was explored via the Tumor Immune Estimation Resource (TIMER). The mechanism underlying the effect of PHF21A on HCC progression was analyzed by gene set enrichment analysis (GSEA) and protein‒protein interaction (PPI) network analysis. Immunohistochemical staining showed that PHF21A expression in HCC tissue was significantly lower than that in adjacent nontumor liver tissue and was associated with patient sex, tumor size, metastasis, and Edmondson grade (p < 0.05). Kaplan-Meier analysis demonstrated that low PHF21A expression was associated with a poor prognosis, and Cox regression analysis showed that PHF21A was an independent predictor of prognosis. TIMER analysis showed that PHF21A is positively correlated with tumor immune cell infiltration levels. Functional annotation indicated that PHF21A is involved in important pathways, including transcriptional deregulation pathways in cancer. Finally, in vitro experiments confirmed the low expression of PHF21A in HCC cells. PHF21A affects the progression and prognosis of HCC, suggesting that PHF21A may play an important role in monitoring and preventing the development of HCC.

Downregulation of Methionine Cycle Genes MAT1A and GNMT Enriches Protein-Associated Translation Process and Worsens Hepatocellular Carcinoma Prognosis.

The major biological methyl donor, S-adenosylmethionine (adoMet) synthesis occurs mainly in the liver. Methionine adenosyltransferase 1A (MAT1A) and glycine N-methyltransferase (GNMT) are two key enzymes involved in the functional implications of that variation. We collected 42 RNA-seq data from paired hepatocellular carcinoma (HCC) and its adjacent normal liver tissue from the Cancer Genome Atlas (TCGA). There was no mutation found in MAT1A or GNMT RNA in the 42 HCC patients. The 11,799 genes were annotated in the RNA-Seq data, and their expression levels were used to investigate the phenotypes of low MAT1A and low GNMT by Gene Set Enrichment Analysis (GSEA). The REACTOME_TRANSLATION gene set was enriched and visualized in a heatmap along with corresponding differences in gene expression between low MAT1A versus high MAT1A and low GNMT versus high GNMT. We identified 43 genes of the REACTOME_TRANSLATION gene set that are powerful prognosis factors in HCC. The significantly predicted genes were referred into eukaryotic translation initiation (EIF3B, EIF3K), eukaryotic translation elongation (EEF1D), and ribosomal proteins (RPs). Cell models expressing various MAT1A and GNMT proved that simultaneous restoring the expression of MAT1A and GNMT decreased cell proliferation, invasion, as well as the REACTOME_TRANSLATION gene EEF1D, consistent with a better prognosis in human HCC. We demonstrated new findings that downregulation or defect in MAT1A and GNMT genes can enrich the protein-associated translation process that may account for poor HCC prognosis. This is the first study demonstrated that MAT1A and GNMT, the 2 key enzymes involved in methionine cycle, could attenuate the function of ribosome translation. We propose a potential novel mechanism by which the diminished GNMT and MAT1A expression may confer poor prognosis for HCC.

Novel low-avidity glypican-3 specific CARTs resist exhaustion and mediate durable antitumor effects against HCC.

Chimeric antigen receptor engineered T cells (CARTs) for HCC and other solid tumors are not as effective as they are for blood cancers. CARTs may lose function inside tumors due to persistent antigen engagement. The aims of this study are to develop low-affinity monoclonal antibodies (mAbs) and low-avidity CARTs for HCC and to test the hypothesis that low-avidity CARTs can resist exhaustion and maintain functions in solid tumors, generating durable antitumor effects. New human glypican-3 (hGPC3) mAbs were developed from immunized mice. We obtained three hGPC3-specific mAbs that stained HCC tumors, but not the adjacent normal liver tissues. One of them, 8F8, bound an epitope close to that of GC33, the frequently used high-affinity mAb, but with approximately 17-fold lower affinity. We then compared the 8F8 CARTs to GC33 CARTs for their in vitro function and in vivo antitumor effects. In vitro, low-avidity 8F8 CARTs killed both hGPC3high and hGPC3low HCC tumor cells to the same extent as high-avidity GC33 CARTs. 8F8 CARTs expanded and persisted to a greater extent than GC33 CARTs, resulting in durable responses against HCC xenografts. Importantly, compared with GC33 CARTs, there were 5-fold more of 8F8-BBz CARTs in the tumor mass for a longer period of time. Remarkably, the tumor-infiltrating 8F8 CARTs were less exhausted and apoptotic, and more functional than GC33 CARTs. The low-avidity 8F8-BBz CART resists exhaustion and apoptosis inside tumor lesions, demonstrating a greater therapeutic potential than high-avidity CARTs.

Association of SOCS1 mRNA expression with hepatitis B virus infection-related hepatocellular carcinoma

Hepatocellular carcinoma (HCC), the predominant form of liver cancer worldwide, can be triggered by a variety of causes such as chronic hepatitis B virus (HBV) infection. The immune response to HBV activates the janus kinase/signal transducers and activators of transcription (JAK/STAT) signaling pathway. Suppressor of cytokine singnaling 1 (SOCS1) is a negative feedback regulator of JAK/STAT pathway. Our study was carried out to evaluate SOCS1 mRNA expression and its correlation with paraclinical characteristics in patients with HBV-related HCC. The SOCS1 mRNA expression level in adjacent non-tumour tissues and tumour liver tissues were determined in 44 patients with HBV infection-associated HCC by real-time RT-PCR. Our results showed that SOCS1 mRNA expression level in adjacent non-tumour tissues were significantly higher than in HCC tissues (p=0.003). High expression of SOCS1 in adjacent non-tumour tissue was observed in patients with single tumour (p=0.024), tumour size ≤5 cm (p=0.011), vascular invasion (p=0.047), and no vascular invasion (p=0.007). Red blood cell counts were positively correlated with SOCS1 gene expression (Spearman’s rho=0.359, p=0.018). Our results suggested that SOCS1expression may be considered as a potential factor involved in the pathogenesis of HCC.

High expression of miR-3682-3p is an unfavorable prognostic factor of hepatocellular carcinoma

To investigate the expression of miR-3682-3p in hepatocellular carcinoma (HCC) and its correlation with clinical parameters and prognosis of HCC. We conducted a bioinformatics analysis of the expression of miR-3682-3p in HCC and its correlation with the patients' survival, and examined its expression in 18 pairs of fresh and 90 pairs of paraffin-embedded HCC and adjacent tissues using real-time fluorescence quantitative PCR and in situ hybridization, respectively. The correlation of miR-3682-3p expression in HCC with the clinical parameters and prognosis of the patients was analyzed. Multivariate regression analysis was used to explore the possibility of miR-3682-3p expression as an independent prognostic factor of HCC. Bioinformatics analysis showed that miR-3682-3p was highly expressed in HCC and significantly correlated with the survival time of HCC patients (χ2=8.793, P < 0.001). The expression of miR-3682-3p was significantly up-regulated in fresh HCC tissues as compared with the adjacent liver tissues (t=3.073, P=0.007). In paraffin-embedded samples, in situ hybridization revealed positive miR-3682-3p expression in the cytoplasm of HCC and adjacent tissues, and its expression was signifcantly up-regulated in HCC tissues (t=2.659, P=0.009). The expression level of miR-3682-3p was significantly correlated with American Joint Commission on Cancer (AJCC; 8th edition) stage (χ2=4.272, P= 0.039), HBV surface antigen status (χ2=5.143, P=0.023), recurrence (χ2=4.593, P=0.032), tumor size (χ2=4.580, P=0.032) and Edmondson Steiner grade (χ2=4.068, P=0.044). Kaplan-Meier analysis showed that a higher expression of miR-3682-3p was associated with a shorter overall survival time (χ2=4.169, P=0.041) and disease-free survival time (χ2=4.078, P=0.043) of the patients. Multivariate analysis suggested that miR-3682-3p expression was an independent predictor of the prognosis of HCC patients. MiR-3682-3p is up-regulated in HCC to serve as a significant factor that contributes to the occurrence and a poor prognosis of HCC.

P-L12 Exploring Pathological Signatures for Predicting Recurrence of Early-stage Hepatocellular Carcinoma Based on Deep Learning

Abstract Background Early-stage hepatocellular carcinoma (HCC) is the ideal indication for liver resection. High recurrence rate limits the radical possibility. Current clinicopathological determinants are insufficient to reliably evaluate the recurrence risk after surgery. To address this global issue, we aimed to use deep learning to explore novel pathological signatures based on histological slides for predicting early-stage HCC recurrence and to evaluate the relationship between histological features and multi-omics information. Methods 576 pathological images collected from 547 patients with BCLC stage 0-A HCC who underwent hepatectomy from 2006 to 2015 were randomly divided into the training cohort and validation cohort. The external validation cohort was composed of 147 TNM I patients from TCGA database. Weakly supervised convolutional neural networks were used to identify six classes of HCC tissues. Pathological signatures were extracted and two novel risk scores were constructed by LASSO Cox to predict recurrence. The forecast performance of the scores and patients' prognosis were evaluated. The relationship between histological score (HS) and immune infiltrating cells was estimated by clustering analysis. Results The classification accuracy of HCC tissue was 94.17%. The C-indexes of histological score in the training, validation and TCGA cohorts were 0.804, 0.739 and 0.708, respectively. Multivariate analysis showed that microvascular invasion (HR = 1.46, 95% CI: 1.09-1.95) and HS (HR = 4.05, 95% CI: 3.40-4.84) were independent risk factors for recurrence-free survival. Patients in HS high-risk group had elevated alpha fetoprotein, worse tumor differentiation and higher proportion of microvascular invasion. HS was positively correlated with the expression of CD14 in adjacent normal liver tissue (P = 0.013), and negatively correlated with the expression of CD8 in tumor (P &amp;lt; 0.001). Conclusions This study established and validated two novel risk scores based on the histological slides using deep learning. HS performed well in recurrence prediction for early-stage HCC patients and indication of important clinicopathological features.

SQSTM1 Expression in Hepatocellular Carcinoma and Relation to Tumor Recurrence After Radiofrequency Ablation

Autophagy is a process that allows the degradation of detrimental components through the lysosome to maintain cellular homeostasis under variable stimuli. SQSTM1 is a key molecule involved in functional autophagy and is linked to different signaling pathways, oxidative responses, and inflammation. Dysregulation of autophagy is reported in a broad spectrum of diseases. Accumulation of SQSTM1 reflects impaired autophagy, which is related to carcinogenesis and progression of various tumors, including hepatocellular carcinoma (HCC). This study investigated SQSTM1 protein expression in HCC and its relation to the clinicopathological features and the likelihood of tumor recurrence after radiofrequency ablation (RFA). This study included 50 patients with cirrhotic HCC of Barcelona Clinic Liver Cancer stages 0/A-B eligible for RFA. Tumor and peritumor biopsies were obtained just prior to local ablation and assessed for tumor pathological grade and SQSTM1 expression by immunohistochemistry. Patients were followed for one year after achieving complete ablation to detect any tumor recurrence. Serum alpha-fetoprotein level (U=149.50, P=0.027∗) and pathological grade of the tumor (χ2=12.702, P=0.002∗) associated significantly with the tumor response to RFA. SQSTM1 expression level was significantly increased in HCC compared to the adjacent peritumor cirrhotic liver tissues (Z=5.927, P<0.001∗). Significant direct relation was found between SQSTM1 expression level in HCC and the pathological grade of the tumor (H=33.789, P<0.001∗). On follow-up, tumor and peritumor SQSTM1 expression levels performed significantly as a potential predictor of the overall survival, but not the disease recurrence. SQSTM1 expression could determine aggressive HCC, even with reasonable tumor size and number, and identify the subset of HCC patients with short overall survival and unfavorable prognosis. SQSTM1 expression could not predict post-RFA intrahepatic HCC recurrence. SQSTM1 may be a potential biomarker and target for the selection of HCC patients for future therapies.

Liver Parenchymal Transection Through Radiofrequency Ablation Using a Radial Probe: Technical Report of a New Modality.

Various techniques have been reported to reduce blood loss during a parenchymal transection, and the radiofrequency ablation (RFA) technique is one of them. Owing to the charring of the adjacent liver tissue and the inability to use the conventional RFA techniques near major vessels, this study used a radial fiber of RFA. This technical report thus describes a technique to perform parenchymal transection using a radial fiber as well as its advantages and disadvantages. A radial fiber dissipates the energy radially and it has the added advantage of placing along the same and perpendicular axis of the liver parenchyma; it has been used in three patients in this study. The total intraoperative blood loss was 30-50 ml during parenchymal transection, and the intraoperative duration was 120-170 min. Bile leak was noted in one patient, which was then managed using the conventional treatment. Through the present technique, the fiber can be used in the vicinity of major blood vessels and necrosis and charring can be prevented. Although radial fiber has some advantages, it remains in the preliminary stage and requires further validation.

Characterization of Androgen Receptor Complex Associated Protein (ARCAP) in hepatocellular carcinoma and liver.

Hepatocellular carcinoma (HCC) ranks many tasks in clinical oncology due to possibly developing a general tumor in men and, usually lead to malignant to death within years. Researches had reported about major factors for being HCC was male sex and HCC associated with cirrhosis in childhood was found more common in males than females. In certain mouse strains as studied, breeding with testosterone significantly increases the development of HCC. Furthermore, castration of male mice diminished the frequency of the development of liver tumors. Meanwhile male hepatitis B virus transgenic mice have a greater occurrence of HCC than females. We apply degenerate priming PCR to observe the expression of various steroid receptors in livers. Yeast-two hybrid screening to search a novel RNA fragment helps to find a new full-length gene by RACE experiment. RT-PCR is applied to detect various expressions in tissues and cell lines. In situ hybridization detects DNA in Chromosome mapping. GFP-constructs transfection proves the gene localization in cells. Immunoprecipitation pulldown assay verifies protein interaction. Gene transfection followed with luciferase assay demonstrates the interaction of genes within cellular signaling. Genomic alignment analysis for observing sequences data perform from NCBI database website (http://www.ncbi.nim.nih.gov/genebank/). The androgen receptor (AR) expression level is found at the highest level among the steroid receptors families detected in liver tumors. By yeast-two hybrid screening, we cloned an Androgen Receptor Complex Associated Protein (ARCAP), of 95 Kd in molecular weight and its cDNA. ARCAP locates at Chromosome 1. Our findings indicate ARCAP is highly expressed in hepatoma cell lines and liver tumors and their adjacent tumors as observed. Yeast two-hybrid assay and in vitro immunoprecipitation assays demonstrated an interaction between AR and ARCAP. We aim to search for different types and levels of steroid receptors expressed within human HCCs and in the adjacent liver tissues. To verify possible molecular mechanisms by which AR might affect hepatoma cells, we had characterized a novel protein ARCAP which functions as a coregulator to interact with AR within liver. The ligand-dependent AR with its cofactor, ARCAP, can induce a signal cascade by transactivation.

Expression of SET domain bifurcated histone lysine methyltransferase 1 and its clinical prognostic significance in hepatocellular carcinoma.

BackgroundTo detect the expression of histone methyltransferase SETDB1 in hepatocellular carcinoma, and to analyze the relationship between SETDB1 expression and tumor size, microvascular invasion, pTNM stage, gender, age, tumor number, tumor differentiation, and other clinicopathological characteristics.MethodsImmunohistochemical method was used to detect the expression of SETDB1 proteins in liver cancer tissues and adjacent tissues of 100 cases. The qRT‐PCR method was used to detect the expression of SETDB1 mRNA in hepatocellular carcinoma and adjacent tissues of 64 cases.ResultsThe expression of SETDB1 protein and mRNA in hepatocellular carcinoma was higher than that of adjacent normal liver tissue (p < 0.05). High protein expression of SETDB1 was associated with tumor size, MVI presence, and pTNM stage (p < 0.05). Univariate analysis revealed that the tumor size, tumor differentiation, MVI grade, and pTNM stage were correlated with DFS, while tumor size, MVI grade, pTNM stage, and SETDB1 protein expression were correlated with OS. Multivariate analysis showed that the combination of MVI grade and pTNM stage has statistical significance in predicting prognosis, while SETDB1 protein expression was not significant prognosis factor.ConclusionsSETDB1 has a certain role in HCC progression and may act as a prognostic predictor concerning the survival of HCC patients.