Hepatocellular Carcinoma Tumor Size Research Articles

Hepatocellular carcinoma with multiple primary malignancy: ARetrospective Study of 106 Cases.

Multiple primary malignancies (MPM) are a rare scenario, particularly in patients with hepatocellular carcinoma (HCC). Research addressing MPM patients with HCC is limited. Therefore, we conducted a retrospective study to explore the clinical features and outcomes of MPM patients involving HCC. We retrospectively analyzed records of patients diagnosed with HCC from January 2013 to October 2023 in the First Affiliated Hospital of Xiamen University.HCC patients with extrahepatic tumors were identified. Their clinical characteristics and survival data were were further analyzed. Among the 1556patients with HCC,106 (6.8%) were identified with EHPM,of which29weresynchronous and 77weremetachronous. A total of 96 patientshad doubleprimary cancers,and10patientshad triple primary cancers. The most common EHPMs were lung cancer (15%), followed by colorectal tumors and stomach cancer. Compared with the synchronous group, the curative treatment rate of HCC and EHPMin the metachronous groupis higher. Duringfollow-up period,29 patients died,of which20 (69%)diedfrom HCC-related causes. The median overall survival (OS) time was 88 months, with 1-, 3-, 5-, and 10-year cumulative survival rates of 92.4%, 88%, 82.5%, and 69.6%, respectively. The 1-, 3-, and 5-year HCC-specific OS (HOS) rates were 81.8%, 75.8%, and 71.9%, respectively. Univariate analysis revealed that Child-Pugh class (B-C), HCC tumor size >5cm, non-radical treatment for HCC or EHPM, HCC recurrence, BCLC staging (C-D), and synchronous appearance were significantly associated with shorter OS and HOS. Factors such as Childs class, tumor size, treatment modality, and synchronous presentation were identified as independent predictors of OS through Cox analysis. Childs class, tumor size, non-radical treatment, BCLC staging, and HCC recurrence were found to be independent factors affecting HOS. The occurrence of extrahepatic primary tumors is not rare, underscoring the importance for oncologists being alerttothedevelopment of secondary tumor development in HCC patients. However, the co-occurrence of other primary tumors alongside HCC does not appear to worsen patient prognosis. Notably, curative resection, where feasible, emerges as a vital factor in extending patient survival.

Ropivacaine combined with sorafenib attenuates hepatocellular carcinoma cell proliferation and metastasis by inhibiting the miR-224/HOXD10 axis.

The development of drug resistance in hepatocellular carcinoma (HCC) cells limits the effectiveness of sorafenib (Sor). However, the regulatory mechanisms underlying the effects of the combination Sor and ropivacaine (Rop) on HCC cells remain unclear. miR-224 and HOXD10 mRNA expression in HCC cells was analyzed using qRT-PCR. CCK-8, Transwell assays and tumor formation experiments in nude mice were used to assess HCC cell proliferation, migration, and invasion. Migration of HCC cells was also analyzed using a cell scratch assay. Hematoxylin and eosin staining was used to detect tumor area. miR-224 expression profoundly increased in HepG2 and Huh7 cells. Treatment with Rop and/or Sor blocked miR-244 expression, especially the combination treatment. Transfection of miR-224 mimic increased HCC cell proliferation and tumor size in nude mice, and migration and invasion in vitro in the presence of Rop and Sor compared to the negative control mimic. Dual-luciferase reporter assays showed that HOXD10 was targeted by miR-224. HOXD10 protein expression and was markedly reduced in HepG2 and Huh7 cells. Rop and/or Sor treatment increased HOXD10 protein expression, particularly the combination treatment. miR-224 negatively regulated HOXD10 expression in HCC cells treated with Rop and Sor. Transfection-mediated silencing of HOXD10 increased HCC cell proliferation, migration, and invasion in the presence of Rop and Sor compared with negative control transfection. The combination of Rop and Sor attenuates HCC cell proliferation and metastasis via the miR-224/HOXD10 axis.

Predictive value of a stemness-based classifier for prognosis and immunotherapy response of hepatocellular carcinoma based on bioinformatics and machine-learning strategies.

Significant advancements have been made in hepatocellular carcinoma (HCC) therapeutics, such as immunotherapy for treating patients with HCC. However, there is a lack of reliable biomarkers for predicting the response of patients to therapy, which continues to be challenging. Cancer stem cells (CSCs) are involved in the oncogenesis, drug resistance, and invasion, as well as metastasis of HCC cells. Therefore, in this study, we aimed to create an mRNA expression-based stemness index (mRNAsi) model to predict the response of patients with HCC to immunotherapy. We retrieved gene expression and clinical data of patients with HCC from the GSE14520 dataset and the Cancer Genome Atlas (TCGA) database. Next, we used the "one-class logistic regression (OCLR)" algorithm to obtain the mRNAsi of patients with HCC. We performed "unsupervised consensus clustering" to classify patients with HCC based on the mRNAsi scores and stemness subtypes. The relationships between the mRNAsi model, clinicopathological features, and genetic profiles of patients were compared using various bioinformatic methods. We screened for differentially expressed genes to establish a stemness-based classifier for predicting the patient's prognosis. Next, we determined the effect of risk scores on the tumor immune microenvironment (TIME) and the response of patients to immune checkpoint blockade (ICB). Finally, we used qRT-PCR to investigate gene expression in patients with HCC. We screened CSC-related genes using various bioinformatics tools in patients from the TCGA-LIHC cohort. We constructed a stemness classifier based on a nine-gene (PPARGC1A, FTCD, CFHR3, MAGEA6, CXCL8, CABYR, EPO, HMMR, and UCK2) signature for predicting the patient's prognosis and response to ICBs. Further, the model was validated in an independent GSE14520 dataset and performed well. Our model could predict the status of TIME, immunogenomic expressions, congenic pathway, and response to chemotherapy drugs. Furthermore, a significant increase in the proportion of infiltrating macrophages, Treg cells, and immune checkpoints was observed in patients in the high-risk group. In addition, tumor cells in patients with high mRNAsi scores could escape immune surveillance. Finally, we observed that the constructed model had a good expression in the clinical samples. The HCC tumor size and UCK2 genes expression were significantly alleviated and decreased, respectively, by treatments of anti-PD1 antibody. We also found knockdown UCK2 changed expressions of immune genes in HCC cell lines. The novel stemness-related model could predict the prognosis of patients and aid in creating personalized immuno- and targeted therapy for patients in HCC.

Evaluation of the efficacy and safety of CalliSpheres® microsphere-transarterial chemoembolization in large hepatocellular carcinoma.

The prognosis of large hepatocellular carcinoma (HCC) is still unfavorable due to limited and challenging treatment. CalliSpheres® microsphere-transarterial chemoembolization (CSM-TACE) is an effective therapy for general HCC but not frequently applied for large HCC. Hence, this study aimed to investigate the efficacy and safety of CSM-TACE in large HCC patients. This prospective study analyzed 100 large HCC (tumor size >5 cm) patients receiving CSM-TACE. Treatment response, survival, change in liver function indexes, and adverse events were recorded. The best complete response, partial response, stable disease, and progressive disease rates were 2.0%, 31.3%, 65.7%, and 1.0%, respectively, leading to the best objective response rate (ORR) of 33.3% and disease control rate of 99.9%. Multivariate analysis showed that intrahepatic metastasis was independently related to poor ORR (odd ratio = 0.366, P = 0.023). The 1- and 2-year progression-free survival (PFS) rates were 88.9% and 80.6%, with a mean [95% confidence interval (CI)] PFS of 21.6 (20.4-22.9) months. The 1- and 2-year overall survival (OS) rates were 99.0% and 99.0%, with a mean (95% CI) OS of 23.8 (23.3-24.2) months. Total bilirubin (P < 0.001), alanine transaminase (P < 0.001), aspartate transaminase (P < 0.001), and α-fetoprotein (P = 0.045) were abnormal in a short-term period then stably recovered from 1 month ± 15 days after drug-eluting bead-TACE to 24 months ± 15 days. During hospitalization and postdischarge, tolerable abdominal pain and decreased appetite were common adverse events. CSM-TACE shows favorable treatment response and survival with acceptable tolerance among large HCC patients, indicating that it may promote the management of these patients.

Development and validation of a diagnostic model for AFP-negative hepatocellular carcinoma.

AFP appears to be negative in about 30% of overall hepatocellular carcinoma (HCC). Our study aimed to develop a nomogram model to diagnose AFP-negative HCC (AFPN-HCC). The training set included 294 AFPN-HCC patients, 159 healthy objects, 63 patients with chronic hepatitis B(CHB), and 64 patients with liver cirrhosis (LC). And the validation set enrolled 137 healthy controls objects, 47 CHB patients and 45 patients with LC. LASSO, univariate, and multivariable logistic regression analysis were performed to construct the model and then transformed into a visualized nomogram. The receiver operating characteristic (ROC) curves, the calibration curve, decision curve analysis (DCA), and clinical impact curve (CIC) were further used for validation. Four variables including age, PIVKA-II, platelet (PLT) counts, and prothrombin time (PT) were selected to establish the nomogram. The area under the curve (AUC) of the ROC to distinguish AFPN-HCC patients was 0.937(95% CI 0.892-0.938) in training set and 0.942(95% CI 0.921-0.963) in validation set. We also found that the model had high diagnostic value for small-size HCC (tumor size < 5cm) (AUC = 0.886) and HBV surface antigen-positive AFPN-HCC (AUC = 0.883). Our model was effective for discrimination of AFPN-HCC from patients with benign liver diseases and healthy controls, and might be helpful for the diagnosis for AFPN-HCC.

Clathrin light chain A facilitates small extracellular vesicle uptake to promote hepatocellular carcinoma progression

BackgroundEndocytosis is a fundamental process for internalizing small extracellular vesicles (sEVs). The present study aimed to elucidate the role of clathrin light chain A (CLTA) in sEV uptake in hepatocellular carcinoma (HCC).Materials and methodsCLTA expression was analyzed by bioinformatics, quantitative PCR and immunohistochemistry. The clinical relevance of CLTA was analyzed by Fisher’s exact test, Kaplan–Meier analysis, and multivariate cox regression model. The functions of CLTA in sEV uptake and cancerous properties were examined by PKH67-sEV uptake, MTT, colony formation, and transwell assays. Mass spectrometry was used to identify the downstream effectors of CLTA. CLTA inhibitor, Pitstop 2, was tested in a mouse model of patient-derived xenografts (PDXs).ResultsCLTA expression was higher in tumor tissues than in non-tumorous liver tissues and progressively increased from the early to late tumor stage. CLTA overexpression was associated with larger tumor size and poor prognosis in HCC. Cellular CLTA contributed to the sEV uptake, resulting in enhanced cancerous properties. Mechanistically, CLTA increases capping actin protein gelsolin-like (CAPG) expression to facilitate sEV uptake, thereby promoting the proliferation, motility, and invasiveness of HCC cells. What’s more, the CLTA inhibitor Pitstop 2 alone or in combination with sorafenib attenuated tumor growth in mice implanted with PDXs.ConclusionsThe study reveals the role of CLTA in sEV uptake to promote HCC progression. Inhibition of CLTA and its mediated pathway illuminate a new therapeutic strategy for HCC patients.

Association between serum midkine levels and tumor size in Indonesian hepatocellular carcinoma patients: a cross-sectional study.

Background: The incidence of liver cancer is increased worldwide with 75%-85% diagnosed as hepatocellular carcinoma (HCC). Current practice has low sensitivity limitations to diagnose the early stages of HCC, thus urging the need for a biomarker with higher sensitivity to detect HCC, specifically in the early stage. This study aimed to determine the association between midkine levels and progressiveness of hepatocellular carcinoma (HCC), according to tumor size, Barcelona Clinic Liver Cancer (BCLC), and presence of portal venous thrombosis. Methods: This cross-sectional study involved 100 patients in Adam Malik General Hospital diagnosed with HCC, collected with a consecutive sampling method, whose diagnoses were confirmed by findings of hypervascular on arterial phase imaging and portal vein or delayed phase washout triple-phase CT Scan. Samples are later categorized according to Barcelona Clinic Liver Cancer (BCLC) stages, tumor size, and presence of portal venous thrombosis. Blood samples were drawn to measure serum midkine using ELISA. Kruskal-Wallis and Mann-Whitney U tests were conducted to determine the difference of midkine levels based on tumor size, BCLC staging, and presence of portal venous thrombosis. Results: Serum midkine level shows a significant difference over tumor size (p=0.014), no significant difference found compared to BCLC stages and presence of portal venous thrombosis. Conclusion: Serum midkine levels are associated with the tumor size of HCC, thus helping physicians determine treatment plans.

Correlation between the complex of small heat shock proteins (HSPBs) and the progression in patients with hepatocellular carcinoma

Small heat shock proteins (HSPBs) regulate various cell functions. We previously reported that HSPB1, HSPB6, and HSPB8 each suppress the progression of hepatocellular carcinoma (HCC). The heterooligomerization of HSPs is speculated to be crucial for functional activities. Here, we investigated the relationship between the complex of HSPBs and the progression of HCC. HSPB1/HSPB6 complex and HSPB1/HSPB8 complex, but not HSPB6/HSPB8 complex, were observed in both HSPB6-overexpressing human HCC-derived HuH-7 cells and resected human HCC tumor tissue. Differentiation, stage, tumor size, and vein invasion of HCC were inversely related to the presence of HSPB complexes in the HCC tissue. Our results strongly suggest that the HSPB complex formation plays a suppressive role in the HCC progression.

Impact of ultrasonographic blind spots for early-stage hepatocellular carcinoma during surveillance.

BackgroundAbdominal ultrasonography (US) is the backbone of hepatocellular carcinoma (HCC) surveillance. Although previous studies have evaluated clinical factors related to surveillance failure, none have focused specifically on US blind spots.MethodsThis study included 1,289 patients who underwent 6 months intervals surveillance using US and serum alpha-fetoprotein (AFP) and were eventually diagnosed with single-nodular HCC. Patients were divided into US-detected group (n = 1,062) and US-missed group (HCC detected only by AFP ≥ 20ng/mL; n = 227). Blind spots consisted of four locations: hepatic dome, caudate lobe or around the inferior vena cava, <1 cm beneath the ribs, and the surface of the left lateral segment. Both groups were compared by HCC location, proportional distribution, treatment method, and overall survival.ResultsA higher proportion of HCCs were located within blind spots in the US-missed group than in the US-detected group (64.3% vs. 44.6%, P < 0.001). HCC ≥ 2 cm detected in blind spots was higher than in non-blind areas (60.3% vs. 47.1%, P = 0.001). Blind spot HCCs were more treated with surgery, whereas those located in a non-blind area were more treated with local ablation. Patients with an HCC located within a blind spot in the US-detected group had better overall survival than the same in the US-missed group (P = 0.008).ConclusionsUsing the current surveillance test, blind spots affected the initially detected HCC tumor size, applicability of the treatment modality, and overall survival. Physicians should pay attention to US blind spots when performing US-based HCC surveillance.

The status of compensated cirrhosis might be negatively associated with the tumor size in patients with hepatitis B virus-related hepatocellular carcinoma

Liver cirrhosis has been a well-known risk factor for the development of hepatocellular carcinoma (HCC). However, this view has recently been challenged. This study aimed to investigate the potential association of cirrhosis with hepatitis B virus (HBV)-related HCC. In this study, two independent multicenter clinical cohorts that included 1,431 HCC patients with chronic HBV infection were retrospectively studied. The first cohort consisted of 334 HCC patients undergoing curative resection and cirrhosis, who have been pathologically diagnosed. The second cohort consisted of 1,087 HCC patients, who have been diagnosed for the presence of cirrhosis based on clinical evidence. Patients of each cohort were further divided into different subgroups according to the presence of cirrhosis and the severity of the cirrhosis. In both cohorts, patients with cirrhosis had smaller tumor size compared to those without cirrhosis (P &lt; 0.05) and a relatively lower proportion of large tumor, defined as tumor size &gt; 5 cm in diameter (P &lt; 0.05). Patients with decompensated cirrhosis had the highest rate of vascular invasion and/or extrahepatic metastases compared with compensated cirrhosis and non-cirrhosis (53.0% vs. 24.8% vs. 26.9%, P &lt; 0.001). In the first cohort, globulin (odds ratio [OR] = 1.096, P = 0.001) and vascular invasion (OR = 4.013, P = 0.013) were independent risk predictors of HCC tumor size &gt;5 cm, while cirrhosis stage Laennec 4B/C was a protective factor (OR = 0.372, P = 0.002). Similar results were observed in the second cohort. In conclusion, this study implied that HCC patients with compensated cirrhosis tend to harbor smaller tumor, but severe cirrhosis favors tumor vascular invasion and metastasis.

LOXL3-promoted hepatocellular carcinoma progression via promotion of Snail1/USP4-mediated epithelial-mesenchymal transition.

Lysyl-oxidase-like 3 (LOXL3) was reported to be essential in epithelial-mesenchymal transition (EMT) of cancers. However, the role of LOXL3 in hepatocellular carcinoma (HCC) remained unclear. In this study, we explored clinical significance, biological functions, and regulatory mechanisms of LOXL3 in HCC. Our study found that LOXL3 expression was markedly associated with the tumor size and clinical stage of HCC, and it was highly expressed in tumor tissues of metastatic HCC patients. High expression of LOXL3 predicted a poor prognosis of HCC. TGF-β1 treatment elevated LOXL3 protein expression and cell invasion, and reduced cell apoptosis in HCC cell lines (SMMC-7721 and Huh-7), while downregulation of LOXL3 reversed the promotive effects of TGF-β1 treatment on LOXL3 protein expression and cell invasion, and the inhibitory effect on cell apoptosis. Mechanistically, LOXL3 interacted with snail family transcriptional repressor 1 (Snail1) through STRING database and RIP assay, and Snail1 bound to ubiquitin-specific peptidase 4 (USP4) promoter by JASPAR database, luciferase reporter gene and Co-IP assays. Overexpression of USP4 reversed the inhibitory effect of LOXL3 silence on EMT in HCC cells through deubiquitinating and stabilizing the expression of Snail1. Moreover, LOXL3-promoted HCC EMT through Wnt/β-catenin/Snail1 signaling pathway. In vivo study revealed that silence of LOXL3-inhibited HCC tumor growth. In conclusion, LOXL3 silence inhibited HCC invasion and EMT through Snail1/USP4-mediated circulation loop and Wnt/β-catenin signaling pathway.

Intended preoperative trans-arterial embolization for large hepatocellular carcinoma: a retrospective cohort study

BackgroundGenerally, a large tumor size of hepatocellular carcinoma (HCC) is associated with poor visibility and uncertainty in the surgical field which results in increased surgical difficulty as well as unfavorable postoperative outcomes. We performed intended preoperative trans-arterial embolization (TAE) in patients with a large HCC. In this study, we investigated the oncological significance of intended preoperative TAE for a large HCC, using a comparison between patients with and without TAE, and detailed analyses for pre- and post-TAE status.MethodsA total of 411 patients who underwent hepatectomy for primary HCC at the University of Yamanashi Hospital between January 2007 and December 2018 were included in this study. The patients were divided into two groups: patients with larger HCCs (≥50 mm, n=51) and those with smaller HCC (<50 mm, n=360) according to the size of their HCCs. Comparison of clinicopathological features between these groups and clinical outcomes between the TAE and non-TAE groups were compared. In addition, a detailed analysis of each case in the TAE group was conducted, comparing clinicopathological factors between pre- and post-TAE status.ResultsThe clinical unfavorable short- and long-term outcomes of patients with large HCCs (≥50 mm) were revealed compared to those with small HCCs (<50 mm). The prognostic analyses showed that a large tumor size and increased tumor markers, multiple tumor numbers, and others were adverse prognostic factors, and vascular invasions and residual tumors were included in the multivariate analysis.Further detailed analyses revealed that the average rates of change in tumor size and tumor shrinkage after TAE were − 48.6±35.6 mm and − 30.7±17.0%, respectively. Pathological high necrotic changes in the tumor, after multiple-times TAE aiming to a better effect, were related to a better prognosis in patients with large HCC. Poor prognostic factors became less common in patients who underwent intended preoperative TAE, and these patients had better prognoses.ConclusionsThe large tumor size of HCC is associated with unfavorable outcomes; the intended preoperative TAE for large HCC patients performed multiple times aiming to affect the tumor as much as possible might improve their prognoses.

Hypoxia-induced LINC00674 facilitates hepatocellular carcinoma progression by activating the NOX1/mTOR signaling pathway.

The hypoxic tumor microenvironment, a fundamental feature of solid tumors, drives hepatocellular carcinoma (HCC) progression through regulating the transcriptional activities of protein-coding and noncoding genes. However, long noncoding RNA (lncRNA)-mediated HCC progression in hypoxic microenvironment remains largely unknown yet. In this study, we found that LINC00674 was upregulated under hypoxic conditions in a HIF-1-dependent manner, and the occupancy of HIF-1 to HRE of LINC00674 gene promoter was essential for its transcription. In addition, LINC00674 level was increased in HCC cell lines and tissues. Clinically, statistical analysis showed that LINC00674 expression was significantly associated with tumor size, venous infiltration, tumor stage and poor prognosis of HCC. Functionally, loss-of-function assays revealed that LINC00674 knockdown inhibited the migration, proliferation and invasion of HCC cells. Furthermore, LINC00674 silencing prominently repressed the mTOR signaling pathway. LINC00674 overexpression-enhanced HCC cell proliferation, migration and invasion were markedly abolished by an mTOR inhibitor rapamycin. NADPH oxidase 1 (NOX1) was positively regulated by LINC00674 in HCC cells. NOX1 knockdown markedly reversed LINC00674-upregulated the p-mTOR level and HCC cells' malignant behaviors. Finally, we found that LINC00674 knockdown attenuated the growth of HCC cells in vivo. Our finding demonstrated that LINC00674 was a new HIF-1 target gene, and hypoxia-induced LINC00674 exerted a pro-proliferative and pro-metastatic role in HCC, possibly by activating the NOX1/mTOR signaling pathway. This study suggested LINC00674 as a promising therapeutic target for HCC.

Persistently Rising Alpha-fetoprotein in the Diagnosis of Hepatocellular Carcinoma: A Review.

Hepatocellular carcinoma (HCC), one of the most common malignant tumors worldwide, is known for its grim prognosis, with untreated life expectancy being only a matter of months after the diagnosis. The difficulty in making a diagnosis early is one of the main contributing factors to the poor prognosis. Alpha-fetoprotein (AFP) had long been used as a surveillance tool, but suboptimal specificity and sensitivity has prompted liver societies to abandon the recommendation for its universal use, even in combination with ultrasonography. Most studies have shown no obvious correlation between serum AFP level and HCC tumor size, stage, or survival post-diagnosis. However, some studies concluded that a gradual rise or persistent elevation in AFP were positive predictors for tumor development. Other studies reported a fall in AFP followed by a rise in patients with HCC as well as persistently rising AFP levels without development of HCC on follow up. Our calculation of the sensitivity and specificity of persistently rising AFP for HCC were both low, at 60% and 35.8%, respectively, indicating that the presence of persistently rising AFP per se did not offer diagnostic benefit. In addition, our calculated mean slopes of persistently rising AFP levels in HCC and non-HCC patients were numerically very different, but the difference was not statistically significant. We conclude that the published data do not support a role for rising AFP levels per se in the diagnosis of HCC.

Upregulation of a novel LncRNA AC104958.2 stabilized by PCBP2 promotes proliferation and microvascular invasion in hepatocellular carcinoma

Long non-coding RNAs (lncRNAs) were reported to be involved in tumorigenesis and progression of hepatocellular carcinoma (HCC). Microvascular invasion (MVI) is an independent predictor for early recurrence and overall survival in postoperative patients with HCC. However, the mechanisms how lncRNAs affect HCC and MVI remain elusive. By RNA sequencing (RNA-seq) in a series of 65 HCC samples and 30 paired adjacent non-tumor liver tissue, we identified a novel lncRNA AC104958.2 that was significantly upregulated in HCC tissues and associated with MVI. Overexpression of AC104958.2 obviously elevated cell viability, metastasis, invasion and epithelial-mesenchymal transition (EMT), while knockout of AC104958.2 mediated by CRISPR/Cas9 technique showed the opposite effects. In addition, the interaction between AC104958.2 and Poly (rC) binding protein 2 (PCBP2) was identified by RNA pull down and mass spectrometry (MS), which was further validated by RNA immunoprecipitation (RIP). PCBP2 was also upregulated in HCC and associated with MVI. High expression of both AC104958.2 and PCBP2 was correlated with tumor size, TNM stage and MVI in HCC. Overexpression of PCBP2 greatly increased the cell viability, metastasis, invasion and EMT. Moreover, actinomycin D assay showed that overexpression of PCBP2 enhanced the RNA stability of AC104958.2. In conclusion, our study showed that a novel lncRNA AC104958.2 exerted oncogenic roles in HCC and might be a promising biomarker and therapeutic target.

Urokinase plasminogen activator predicts poor prognosis in hepatocellular carcinoma.

Hepatocellular carcinoma (HCC) is one of the most common malignancies worldwide and its prognosis remains dismal. Hence, it is important to identify the diagnostic and prognostic biomarkers for HCC. Urokinase plasminogen activator (uPA), an extracellular matrix (ECM)-degrading protease, plays a pivotal role in the invasion and metastasis of HCC. To confirm the clinical significance of uPA in HCC, we explored uPA expression in HCC in The Cancer Genome Atlas (TCGA) database. The expression level of uPA was further verified by quantitative reverse transcription polymerized chain reaction (qRT-PCR) in 133 pairs of primary HCC samples. A survival analysis was conducted with the Kaplan-Meier method in the HCC samples and TCGA database. Our results showed that uPA was overexpressed in HCC and was significantly associated with HCC tumor size (P=0.015), differentiation grade (P=0.028), and absence of tumor encapsulation (P=0.010). Patients with high uPA expression levels had a poor outcome (P=0.026). TCGA database analysis was also consistent with our experimental results. In conclusion, our findings revealed that uPA was overexpressed in HCC and was related to HCC malignant features including tumor size, differentiation grade and absence of tumor encapsulation. High uPA expression had a shorter survival time. It is a potential prognostic biomarker of HCC.

Alternatively Spliced Tissue Factor Levels in the Plasma of Patients with Liver Fibrosis, Liver Cirrhosis, and Hepatocellular Carcinoma: A Single-Center Retrospective Study

Background:Chronic liver diseases (CLD) such as chronic viral hepatitis, metabolic liver diseases, alcoholic liver disease, primary and secondary hemochromatosis, Wilson's disease, and cholestatic or autoimmune liver diseases are the major cause of morbidity and mortality worldwide ranking as the 12th leading cause of overall mortality. CLD induce liver fibrosis characterized by the accumulation of extracellular matrix proteins, such as collagen, in the liver interstitial space. If untreated, liver fibrosis may evolve to end stage cirrhosis, which is associated with a high risk of developing hepatocellular carcinoma (HCC) and liver failure.CLD is accompanied by changes in the coagulation system that embrace not only hypo-, but also hyper-coagulability. In this rebalanced situation, hypercoagulability can contribute, like genetic predisposition or environmental factors, to the progression of fibrosis and complications of cirrhosis.Full-length tissue factor (TF), a glycosylated transmembrane protein, is the only initiator of the extrinsic coagulation cascade in vivo. It is elevated in patients with pancreatic cancer, blood disorders, diabetes, and cardiovascular diseases. During TF pre-mRNA processing, exon 5 can be spliced out, resulting in a frameshift that yields to a soluble TF isoform called alternatively spliced TF (asTF). While its involvement in thrombosis and/or hemostasis is still debated, asTF induces angiogenesis and was found highly expressed in pancreatic ductal adenocarcinoma and promoted cancer progression. More recently, it was reported that asTF mRNA expression levels may constitute a prognostic marker in human gastric cancer.MethodsIn a single-center study, we analyzed retrospectively asTF plasma levels in healthy subjects, patients with liver fibrosis, liver cirrhosis, and HCC. asTF plasma levels were measured using a sandwich enzyme-linked immunosorbent assay (ELISA). In addition, immunostaining for asTF was performed in HCC tissue biopsies using an anti-asTF specific antibody. Values were expressed as mean±SD.ResultsThe patients with HCC displayed the highest mean asTF levels (n=128, 552.2±469.7pg/ml), followed by those with liver fibrosis (n=70, 524.4±453.9pg/ml) and liver cirrhosis (n=62, 357.2±389.8pg/ml). There was a significant difference in asTF levels between healthy subjects (n=60, 127.3±136.3pg/ml) and patients with liver fibrosis (p<0.0001), healthy subjects and cirrhotic patients (p<0.0001), healthy subjects and HCC patients (p<0.0001), cirrhotic patients and HCC patients (p=0.004) and between fibrotic and cirrhotic patients (p=0.03). Between fibrotic patients and HCC patients, no significant difference in asTF levels was detectable (p=0.70). There were no effects of gender, age and underlying liver disease (hepatitis B and C, alcohol abuse, nonalcoholic steatohepatitis) in each patient group on asTF levels. Furthermore, asTF levels did not correlate with Child-Pugh score, grade of liver fibrosis, and HCC tumor size. Preliminary results indicate that asTF is expressed in liver tissue by hepatocytes, cholangiocytes, and endothelial cells.ConclusionOur study revealed that asTF plasma levels were elevated not only in patients with HCC, but also in patients with cirrhosis and even in those with liver fibrosis. This suggests that asTF is produced during fibrotic remodeling of the liver. The lack of correlation between grade of liver fibrosis and asTF levels could be due to the fact that asTF levels did not correlate with the amount of fibrosis deposited in the parenchyma but with the process of producing the fibrosis. In summary, our observations suggest that asTF might play a role in hepatic fibrogenesis. DisclosuresNo relevant conflicts of interest to declare.

B-Cell Receptor-Associated Protein 31 Promotes Metastasis via AKT/β-Catenin/Snail Pathway in Hepatocellular Carcinoma.

Hepatocellular carcinoma (HCC) is one of the most lethal cancer worldwide, characterized with high heterogeneity and inclination to metastasize. Emerging evidence suggests that BAP31 gets involved in cancer progression with different kinds. It still remains unknown whether and how BAP31 plays a role in HCC metastasis. Epithelial–mesenchymal transition (EMT) has been a common feature in tumor micro-environment, whose inducer TGF-β increased BAP31 expression in this research. Elevated expression of BAP31 was positively correlated with tumor size, vascular invasion and poor prognosis in human HCC. Ectopic expression of BAP31 promoted cell migration and invasion while BAP31 knockdown markedly attenuated metastatic potential in HCC cells and mice orthotopic xenografts. BAP31 induced EMT process, and enhanced the expression level of EMT-related factor Snail and decreased contents and membrane distribution of E-cadherin. BAP31 also activated AKT/β-catenin pathway, which mediated its promotional effects on HCC metastasis. AKT inhibitor further counteracted the activated AKT/β-catenin/Snail upon BAP31 over-expression. Moreover, silencing Snail in BAP31-overexpressed cells impaired enhanced migratory and invasive abilities of HCC cells. In HCC tissues, BAP31 expression was positively associated with Snail. In conclusion, BAP31 promotes HCC metastasis by activating AKT/β-catenin/Snail pathway. Thus, our study implicates BAP31 as potential prognostic biomarker, and provides valuable information for HCC prognosis and treatment.

The Mortality and Overall Survival Trends of Primary Liver Cancer in the United States.

Recent trends of hepatocellular carcinoma (HCC) mortality and outcome remain unknown in the United States. We investigated the recent trends of primary liver cancer (excluding intrahepatic cholangiocarcinoma) mortality and HCC stage, treatment, and overall survival (OS) in the United States. The National Center for Health Statistics Database was analyzed to investigate the trend of primary liver cancer mortality. We analyzed the Surveillance, Epidemiology, and End Results 18 Database to assess the temporal trend of tumor size, stage, treatment, and OS of HCC. We investigated the association between HCC diagnosis year and OS using Cox regression analysis. All statistical tests were 2-sided. During 2000-2018, liver cancer mortality rates increased until 2013, plateaued during 2013-2016 (annual percent change = 0.1%/y, 95% confidence interval [CI] = -2.1%/y to 2.4%/y, P = .92), and started to decline during 2016-2018 (annual percent change = -1.5%/y, 95% CI = -3.2%/y to 0.2%/y, P = .08). However, mortality continues to increase in American Indian and Alaska Native, individuals aged 65 years or older, and in 33 states. There was a 0.61% (95% CI = 0.53% to 0.69%, P < .001) increase in localized stage HCC and a 0.86-mm (95% CI = -1.10 to -0.62 mm, P < .001) decrease in median tumor size per year. The 1-year OS rate increased from 36.3% (95% CI = 34.3% to 38.3%) to 58.1% (95% CI = 56.9% to 59.4%) during 2000-2015, and the 5-year OS rate almost doubled from 11.7% (95% CI = 10.4% to 13.1%) to 21.3% (95% CI = 20.2% to 22.4%) during 2000-2011. Diagnosis year (per year) (adjusted hazard ratio = 0.96, 95% CI = 0.96 to 0.97) was independently associated with OS in multivariable analysis. Primary liver cancer mortality rates have started to decline in the United States with demographic and state-level variation. With an increasing detection of localized HCC, the OS of HCC has improved over the past decades.

Correlation of serum alpha fetoprotein (AFP) and tumor size of hepatocellular carcinoma (HCC) in a tertiary care hospital of Peshawar

Introduction: Early diagnosis of Hepatocellular Carcinoma (HCC) by imaging and biochemical tests would be valuable in controlling morbidity and mortality associated with this most common primary hepatic malignancy which is the third leading cause of global cancer-related deaths. Objective: To determine an association between the level of serum Alpha Fetoprotein (AFP) and the size of the tumor in Hepatocellular Carcinoma (HCC).Materials &amp; Methods: This cross-sectional descriptive study was conducted in Rehman Medical Institute, Peshawar, Pakistan from January 2018 till December 2019. Data were gathered from the medical records of Endoscopy Unit of Rehman Medical Institute. Patients were separated into three groups on the basis of serum AFP levels: Group I had standard levels of AFP (&lt;20 IU/ml), Group II had moderately raised AFP (20-399 IU/ml), and Group III had significantly raised AFP (&gt;400 IU/ml). Patients were also separated into three groups on the basis of tumor; in Group A tumor size was less than 3 cm, in Group B it was between 3 and 5 cm, and in Group C the size was greater than 5 cm. For categorical variables, the Chi-square and Z test for proportions were applied; the Wilcoxon Rank-Sum (Mann-Whitney) test was used to evaluate the variation in the two categories, and the Kruskal-Wallis Rank test was used to evaluate the difference between categories. A p value ≤0.05 was taken as significant. Descriptive statistics were obtained by SPSS 24. Result: A total of 136 patients were evaluated according to AFP levels. The male to female ratio was 2.31:1. The mean presenting age was 59.36 ± 9.85 years while the mean tumor size was 6.93 ± 4.39 cm. Regarding AFP groups, there were 50(36.76%), in Group I, 31(22.79%) in Group II and 38(27.94%) in Group III. As per the Child Pugh classification, Class A, B and C were 69.85%, 16.18% and 0% respectively.Conclusion: Serum alpha fetoprotein was significantly associated with HCC tumor size. Alpha fetoprotein level can be a helpful marker to detect HCC and to differentiate between its initial and late stages. Monitoring AFP levels could be useful to diagnose recurring disease for patients with tumors that produce AFP.Keywords: Serum Alpha Fetoprotein; Hepatocellular Carcinoma; Tumor size.