Prognostic significance of TNFRSF4 expression and development of a pathomics model to predict expression in hepatocellular carcinoma.

Background: TNFRSF4 plays a significant role in cancer progression, especially in hepatocellular carcinoma (HCC). This study aims to investigate the prognostic value of TNFRSF4 expression in patients with HCC and to develop a predictive pathomics model for its expression. Methods: A cohort of patients with HCC retrieved from the TCGA database was analyzed using RNA-seq analysis to determine TNFRSF4 expression and its impact on overall survival (OS). Additionally, hematoxylin-eosin staining analysis was performed to construct a pathomics model for predicting TNFRSF4 expression. Then, pathway enrichment analysis was conducted, immune checkpoint markers were investigated, and immune cell infiltration was examined to explore the underlying biological mechanism of the pathomics score. Results: TNFRSF4 expression was significantly higher in tumor tissues than in normal tissues. TNFRSF4 expression also exhibited significant correlations with various clinical variables, including pathologic stage III/IV and R1/R2/RX residual tumor. Furthermore, elevated TNFRSF4 expression was associated with unfavorable OS. Interestingly, in the subgroup analysis, elevated TNFRSF4 expression was identified as a significant risk factor for OS in male patients. The newly developed pathomics model successfully predicted TNFRSF4 expression with good performance and revealed a significant association between high pathomics scores and worse OS. In male patients, high pathomics scores were also associated with a higher risk of mortality. Moreover, pathomics scores were also involved in specific hallmarks, immune-related characteristics, and apoptosis-related genes in HCC, such as epithelial-mesenchymal transition, Tregs, and BAX expression. Conclusions: Our findings suggest that TNFRSF4 expression and the newly devised pathomics scores hold potential as prognostic markers for OS in patients with HCC. Additionally, gender influenced the association between these markers and patient outcomes.

Family with sequence similarity 111 member B contributes to tumor growth and metastasis by mediating cell proliferation, invasion, and EMT via transforming acidic coiled-coil protein 3/PI3K/AKT signaling pathway in hepatocellular carcinoma.

As a complex systemic disease, primary liver cancer ranks third in death rate for solid tumors worldwide. Family with sequence similarity 111 member B (FAM111B), which was found to be aberrantly mutated in multiple cancers, is a candidate oncogene. We aimed to determine the function and mechanism of FAM111B in hepatocellular carcinoma (HCC). The expression of FAM111B was evaluated in HCC tissues, adjacent tissues, HCC cell lines. The impact of FAM111B on proliferation, invasion, apoptosis and EMT of HCC cells were detected by CCK-8, Transwell, flow cytometry and Western blot assays. The relationship between FAM111B and transforming acidic coiled-coil protein 3 (TACC3) was assessed by CoIP and Immunofluorescence (IF) staining assays. The effect of FAM111B on tumor growth was detected by using xenograft model of nude mice. The expression of FAM111B was upregulated in HCC tissues and cell lines, and the prognosis of HCC patients was worse in the high FAM111B expression group, and its expression level was associated with the TNM stage of HCC. FAM111B silencing inhibited HCC cell proliferation and invasion, EMT and induced apoptosis. Besides, TACC3 served as an interactor for FAM111B, which could enhance TACC3 expression, thus activing PI3K/AKT pathway. Rescue experiments revealed that elevated of TACC3 restored the inhibitory effect of FAM111B overexpression on the cell functions via PI3K/AKT pathway. In vivo, FAM111B inhibition hampered tumor growth and metastasis of HCC. This study highlighted a key player of FAM111B in modulating the malignant biological progression of HCC via TACC3/PI3K/AKT signaling pathway, displaying a potential therapeutic target for HCC.

Different percutaneous transhepatic biliary stent placements and catheter drainage in the treatment of middle and low malignant biliary obstruction.

BACKGROUND: For cases of middle and low biliary obstruction with left and right hepatic duct dilatation, the type of approach and whether different approaches affect the difficulty of puncture operation and intraoperative and postoperative complications have not been discussed in detail. AIM: To compare the efficacy of different percutaneous transhepatic biliary stent placements and catheter drainage in treating middle and low biliary obstruction. METHODS: A retrospective analysis was performed on the medical records of 424 patients with middle and low biliary obstruction who underwent percutaneous liver puncture biliary stent placement and catheter drainage at the Department of Interventional Radiology, Shaanxi Provincial People's Hospital between March 2016 and March 2022. Based on the puncture path, patients were categorized into two groups: Subxiphoid left hepatic lobe approach group (Group A, 224 cases) and right intercostal, right hepatic lobe approach group (Group B, 200 cases). Liver function improvement, postoperative biliary bleeding incidence, postoperative pain duration, and abdominal effusion leakage around the drainage tube were compared between the two groups at 3 d and 1 wk after the surgery. Patient survival time was recorded during follow-up. RESULTS: All 424 surgeries were successful without adverse events. Group A comprised 224 cases, and Group B had 200 cases. There was no statistically significant difference in basic data between Group A and Group B (P > 0.05). No significant difference in postoperative biliary bleeding incidence was observed between the groups (P > 0.05). The decreased rates for total bilirubin (Group A: 69.23 ± 4.50, Group B: 63.79 ± 5.65), direct bilirubin (Group A: 79.30 ± 11.19, Group B: 63.62 ± 5.64), and alkaline phosphatase (Group A: 60.51 ± 12.23, Group B: 42.68 ± 23.56) in the 1st wk after surgery were significantly faster in Group A than in Group B. The decreased rate of gamma-glutamyl transpeptidase was also significantly faster in Group A at both 3 d (Group A: 40.56 ± 10.32, Group B: 32.22 ± 5.12) and 1 wk (Group A: 73.19 ± 7.05, Group B: 58.81 ± 18.98) after surgery (P < 0.05). Group A experienced significantly less peritoneal effusion leakage around the drainage tube than Group B (P < 0.05). The patient survival rate was higher in Group A compared to Group B (P < 0.05). CONCLUSION: In treating jaundice patients with middle and low biliary obstruction, a percutaneous left liver puncture demonstrated better clinical efficacy than a percutaneous right liver puncture.

Clinical advantage of drug-coated balloon in treatment of restenosis in superficial femoral artery stents.

BACKGROUND: This investigation probed the clinical effectiveness of drug-coated balloon (DCB) and common balloon (CB) for treating restenosis in superficial femoral artery stents. METHODS: We retrospectively analyzed 295 patients with lower extremity arteriosclerosis obliterans with superficial femoral artery stenting restenosis admitted to Shaanxi Provincial People's Hospital from March 2017 to March 2020. Cases were segregated within two cohorts adopting different treatment methods (cohort 1, DCB and cohort 2, CB). The restenosis and clinical outcomes were evaluated by Doppler ultrasound, computed tomography angiography, or digital subtraction angiography at 1 week, 6, and 12 months. RESULTS: Following six months, the restenosis rate on angiography was 4.2% within cohort 1, and 26% within cohort 2 (P<0.05); at 12 months the rates on duplex ultrasonography were 18% and 84%, respectively (P<0.001). ABI and MDL of cohort 1 (P<0.05) were higher than those of cohort 2, 6 and 12 months post-operation; late lumen loss (LLL) and peak systolic velocity ratio (PSVR) of cohort 1 (P<0.05) were lower than those of Cohort 2. Cohort 1 cases demonstrated increased mobility upon treadmill at 12 months compared to within cohort 2. CONCLUSIONS: Medium-/long-term clinical effectiveness for drug-coated balloon treating restenosis in a superficial femoral artery stent is significantly higher than that of an ordinary balloon.

Construction and Validation of Prediction Model of Severe Abdominal Pain Post-Transarterial Chemoembolization in Patients with HBV-Associated Primary Liver Cancer.

Objective: This study is aimed at constructing and evaluating a prediction model of severe abdominal pain post-transcatheter arterial chemoembolization in patients with HBV-related primary liver cancer. Methods: Patients with HBV-associated primary liver cancer who received transarterial chemoembolization (TACE) from March 2019 to March 2022 in the Interventional Therapy Department of our hospital were selected as the subjects, and the included 160 patients were randomly divided into modeling group (n = 120) and validation group (n = 40) in a ratio of 3 : 1. Visual analog scale (VAS) was used to assess pain severity. 120 patients in the modeling group were divided into no/mild abdominal pain group and severe abdominal pain group. The clinical data of the patients, including gender, age, TACE treatment history, vascular invasion, maximum diameter of tumor, infarction degree, preoperative Eastern Oncology Collaboration Group (ECOG) score, and Lipiodol dosage, were analyzed. Receiver operating characteristic (ROC) curve was used to evaluate the predictive value of the prediction model for severe abdominal pain post-TACE. Results: A total of 116 patients (72.50%) had severe abdominal pain after TACE. Univariate analysis showed that severe abdominal pain after TACE in the modeling group was associated with TACE treatment history, maximum tumor diameter, infarction degree, and preoperative ECOG score (all P < 0.05), but not related to gender, age, vascular invasion, and Lipiodol dosage (all P > 0.05). Logistic regression analysis showed that TACE treatment history, maximum tumor diameter, infarction degree, and preoperative ECOG score were all independent influencing factors for acute abdominal pain post-TACE in HBV-HCC patients (all P < 0.05). The prediction model equation was Y = -3.673 + 1.722 × TACE treatment history + 1.175 × tumor maximum diameter + 2.064 × infarction degree + 1.555 × preoperative ECOG score. Goodness-of-fit test results showed no significant difference between the established prediction model and the observed value (χ 2 = 1.645, P = 0.560) and R 2 = 0.821, suggesting that the prediction ability of the model was relatively accurate. ROC analysis results showed that the area under the curve (AUC) of severe abdominal pain after TACE was 0.916 (0.862~0.970) and 0.902 (95% CI: 0.841~0.963) in the modeling group and the verification group, respectively. Conclusion: TACE treatment history, tumor maximum diameter, infarction degree, and preoperative ECOG score are independent influencing factors for severe abdominal pain post-TACE in patients with HBV-HCC, and the prediction model established on this basis has good application value.

Efficacy of Drug-Eluting Bead Transarterial Chemoembolization in the Treatment of Colorectal Cancer Liver Metastasis.

We aimed to investigate the efficacy and safety of drug-eluting bead transarterial chemoembolization (DEB-TACE) in the treatment of colorectal cancer liver metastasis. A total of 120 patients with colorectal cancer liver metastasis were divided into the TACE group (receiving TACE treatment, n = 60) and the DEB-TACE group (receiving DEB-TACE treatment, n = 60). At 1 month after treatment, the objective response rate (ORR) in the TACE group and DEB-TACE group were 65.0% (39/60) and 78.3% (47/60), respectively, and the disease control rate (DCR) was 78.3% (47/60) and 85.0% (51/60), respectively. Three months later, the ORRs in TACE and DEB-TACE groups were 63.3% (38/60) and 75.0% (45/60), and the DCRs were 76.7% (46/60) and 81.7% (49/60). We showed that the 1-year overall survival (OS) in TACE and DEB-TACE groups were 100% (60/60) and 88.3% (53/60), respectively, and the 2-year OS were 78.3% (47/60) and 61.7% (37/60). Further analysis indicated that the OS in the DEB-TACE group was significantly longer than that in the TACE group (P = 0.045). DEB-TACE is effective, safe, and feasible in the treatment of colorectal cancer liver metastasis, which can effectively improve the survival of patients.

Capecitabine metronomic chemotherapy combined with autologous CIK cell immunotherapy in the treatment of recurrent and metastatic triple-negative breast cancer.

PURPOSE: The purpose of this study was to investigate the efficacy and safety of maintenance therapy of capecitabine metronomic chemotherapy combined with autologous cytokine-induced killer (CIK) cell immunotherapy in patients with recurrent metastatic triple-negative breast cancer (mTNBC). METHODS: The clinical data of 110 patients with recurrent mTNBC were retrospectively analyzed. Among, 55 were treated with maintenance therapy of capecitabine metronomic chemotherapy combined with autologous CIK cell immunotherapy (DC-CIK group), while the rest 55 were treated with simple metronomic chemotherapy (control group). RESULTS: The ORR of patients in DC-CIK group and control group was 29.1% and 16.4%, and the DCR was 74.5% and 63.6%, respectively. After treatment, the proportions of CD3+ T lymphocytes, CD4+ T lymphocytes and NK cells as well as the CD4/CD8 cell ratio were notably higher in DC-CIK group than those in control group, while the proportion of CD8+ T lymphocytes was notably lower in DC-CIK group than that in control group. Compared with those before treatment, the scores of quality of life evaluated using the FACT-B-V4.0 scale were remarkably improved in both groups after treatment. The score of emotional status and total score were distinctly higher in DC-CIK group than those in control group. Moreover, the follow-up results together with log-rank test revealed that the PFS in DC-CIK group was notably superior to that in control group. CONCLUSIONS: The maintenance therapy of capecitabine metronomic chemotherapy combined with DC-CIK cell immunotherapy is effective in the treatment of recurrent mTNBC, with tolerable adverse reactions. It can improve the patients' immune function, improve their quality of life, and prolong their PFS. Key words: capecitabine, metronomic chemotherapy, immunotherapy, breast cancer, recurrent and metastatic.

Circadian clock gene NPAS2 promotes reprogramming of glucose metabolism in hepatocellular carcinoma cells.

Emerging evidences show that dysregulation of circadian genes is closely associated with tumorigenesis. However, whether circadian genes regulate the reprogramming of metabolism in tumor cells is largely unknown. Here, we showed that NPAS2, one of the core circadian molecules, significantly contributed to the reprogramming of glucose metabolism mainly through two mechanisms. On the one hand, NPAS2 upregulated the expression of glycolytic genes GLUT1, HK2, GPI, ALDOA, ENO2, PKM2 and MCT4. On the other hand, NPAS2 downregulated the expression of peroxisome proliferator-activated receptor gamma coactivator 1 alpha (PGC-1α). Mechanistically, HIF-1α was found to be a direct transcriptional target of NPAS2, which mediated both the upregulation of glycolytic genes and downregulation of mitochondrial biogenesis in HCC cells. In addition, we found that upregulation of NPAS2 expression was mainly due to the downregulation of miR-199b-5p. In vitro and in vivo assays further indicated that HIF-1α-mediated reprogramming of glucose metabolism played a critical role in NPAS2-regulated growth and metastasis of HCC cells. Our findings demonstrate that NPAS2 plays a critical role in glucose metabolism reprogramming, suggesting that NPAS2 may serve as a potential therapeutic target in HCC.

NPAS2 Contributes to Liver Fibrosis by Direct Transcriptional Activation of Hes1 in Hepatic Stellate Cells.

Recently, emerging evidence shows that dysregulation of circadian genes is closely associated with liver fibrosis. However, how dysregulation of circadian genes promotes liver fibrosis is unknown. In this study, we show that neuronal PAS domain protein 2 (NPAS2), one of the core circadian molecules that has been shown to promote hepatocarcinoma cell proliferation, significantly contributed to liver fibrogenesis. NPAS2 is upregulated in hepatic stellate cells (HSCs) after fibrogenic injury, which subsequently contributes to the activation of HSCs. Mechanistically, NPAS2 plays a profibrotic role via direct transcriptional activation of hairy and enhancer of split 1 (Hes1), a critical transcriptor of Notch signaling for the fibrogenesis process, in HSCs. Our findings demonstrate that NPAS2 plays a critical role in liver fibrosis through direct transcriptional activation of Hes1, indicating that NPAS2 may serve as an important therapeutic target to reverse the progression of liver fibrosis.

[Elevated expression of CLOCK is associated with poor prognosis in hepatocellular carcinoma].

Objective To evaluate the expression of circadian locomotor output cycles kaput (CLOCK) and its effects on cell growth in hepatocellular carcinoma (HCC). Methods The expression of CLOCK in 158 pairs of human HCC tissues and matched noncancerous samples was detected by immunohistochemical (IHC) staining. The expression of CLOCK in HCC patients was also verified using the data from GEO and TCGA (a total of 356 cases). The relationship between CLOCK expression and clinicopathological features of HCC patients was analyzed by single factor statistical analysis. Kaplan-Meier survival curves of HCC patients were drawn to study the relationship between the expression level of CLOCK and the survival state. The effect of CLOCK on the growth of HepG2 cells was detected by MTS assay. Results The expression of CLOCK in HCC tissues was significantly higher than that in the adjacent tissues, and the up-regulation of CLOCK expression in HCC tissue was also confirmed in the public data of HCC (356 cases). HCC patients were divided into low CLOCK expression group and high CLOCK expression group. Univariate analysis showed that the expression of CLOCK was related to tumor size, TNM stage, and portal vein invasion in HCC patients. HCC patients with low CLOCK expression had longer overall survival time and relapse-free survival time than those with high CLOCK expression. The proliferation of cells significantly decreased after the expression of CLOCK was knocked down in HepG2 cells. Conclusion The expression of CLOCK in HCC tissues was much higher than that in normal liver tissues, and the high expression of CLOCK indicated the poor prognosis. The knockdown of CLOCK in HCC cells could inhibit the proliferation of HepG2 cells.

Genetic variants in glucose-6-phosphate isomerase gene as prognosis predictors in hepatocellular carcinoma.

BACKGROUND: Metabolic reprogramming is an important hallmark of cancer cells, including the alterations of activity and expression of enzymes in glucose metabolism. Previous studies have demonstrated the critical role of glucise-6-phosphate isomerase (GPI) in cancer initiation, metastasis and progression. However, the significance of single nucleotide polymorphisms (SNPs) in GPI gene has not been investigated in hepatocellular carcinoma (HCC). METHODS: In this study, a total of 3 functional SNPs in GPI gene were genotyped in 492 HCC patients with surgical treatment. Multivariate Cox proportional hazards model and Kaplan-Meier curve were used for the analysis of overall survival (OS) and recurrence-free survival (RFS). RESULTS: The homozygous variant genotypes of rs7248411 in mRNA splice sites of GPI gene were significantly associated with an increased risk of death in the multivariate analysis (Hazard ratio [HR], 2.07; 95% confidence interval [95% CI]: 1.16-3.68 in a recessive model). In stratified analysis, the association remained significant in patients with high α-fetal protein (AFP) level (HR=2.37, 95% CI 1.25-4.49). Moreover, we identified the interaction between rs7248411 and AFP level in predicting the prognosis of HCC patients (P for interaction<0.001). CONCLUSIONS: Our data suggest that GPI gene polymorphism may serve as potential biomarkers to predict the OS of HCC. Further studies with different ethnicities are needed to validate our findings and generalize its clinical utility.

CD147 promotes liver fibrosis progression via VEGF-A/VEGFR2 signalling-mediated cross-talk between hepatocytes and sinusoidal endothelial cells.

Although previous evidence indicates close involvement of CD147 in the pathogenesis of liver fibrosis, the underlying molecular mechanisms and its therapeutic value remain largely unknown. In the present study, we investigated the biological roles of CD147 in liver fibrosis and assessed its therapeutic value as a target molecule in the CCl4-induced liver fibrosis mouse model. We found that CD147 was highly expressed in both hepatocytes and SECs (sinusoidal endothelial cells) in fibrotic liver tissues. Additionally, it was significantly associated with the fibrosis stage. TGF-ß1 (transforming growth factor ß1) was found to be mainly responsible for the up-regulation of CD147. Bioinformatic and experimental data suggest a functional link between CD147 expression and VEGF-A (vascular endothelial growth factor A)/VEGR-2 (VEGF receptor 2) signalling-mediated angiogenesis in fibrotic liver tissues. Furthermore, we observed that the CD147-induced activation of the PI3K (phosphoinositide 3-kinase)/Akt signalling pathway promotes the production of VEGF-A in hepatocytes and expression of VEGFR-2 in SECs, which was found to enhance the angiogenic capability of SECs. Finally, our data indicate that blocking of CD147 using an mAb (monoclonal antibody) attenuated liver fibrosis progression via inhibition of VEGF-A/VEGFR-2 signalling and subsequent amelioration of microvascular abnormality in the CCl4-induced mouse model. Our findings suggest a novel functional mechanism that CD147 may promote liver fibrosis progression via inducing the VEGF-A/VEGFR-2 signalling pathway-mediated cross-talk between hepatocytes and SECs. New strategies based on the intervention of CD147 can be expected for prevention of liver fibrosis.

Transarterial chemoembolization aggravated peritumoral fibrosis via hypoxia-inducible factor-1α dependent pathway in hepatocellular carcinoma.

BACKGROUND AND AIM: It was commonly accepted that chemotherapeutic cytotoxicity was the main cause for hepatic failure in hepatocellular carcinoma patients after repeated transarterial chemoembolization (TACE). However, the effect of embolization-induced hypoxia on liver cirrhosis has rarely been concerned. METHODS: Serum levels of alanine aminotransferase, aspartate aminotransferase, and albumin were used to detect liver injury. Hepatic artery ligation was performed in carbon tetrachloride-induced rat hepatic fibrosis model to mimic the effect of hepatic hypoxia on liver fibrosis after TACE. Sirius Red staining and immunohistochemical analysis of alpha-smooth muscle actin (α-SMA) were used to detect the activation of hepatic stellate cells. Moreover, the expression of hypoxia and fibrosis-related molecules were analyzed at protein and/or mRNA level. RESULTS: Patients showed a significant increase in alanine aminotransferase and aspartate aminotransferase (P = 0.006), accompanied by a decrease in albumin (P = 0.005) after repeated TACE. Hepatic artery ligation significantly promoted carbon tetrachloride-induced rat liver fibrosis progression as indicated by Sirius Red and α-SMA staining, as well as increased expression of hypoxia-inducible factor (HIF)-1α, transforming growth factor (TGF)-ß1, and vascular endothelial growth factor (VEGF). Conditioned media of hypoxia-treated L02 cells induced the expression of Collagen I and α-SMA in LX-2 cells, which was inhibited by HIF-1α small interfering RNA. Finally, HIF-1α inhibitor LW6 attenuated the hypoxia-induced fibrosis progression in vivo. CONCLUSION: Our data demonstrate that TACE-induced hepatic hypoxia aggravates the fibrosis progression in peritumoral liver tissue, thus leads to the deterioration of liver function. Intervention of HIF-1α might be a valuable strategy to optimize the efficacy and reduce the complication of TACE.

New Insight into the Anti-liver Fibrosis Effect of Multitargeted Tyrosine Kinase Inhibitors: From Molecular Target to Clinical Trials.

Tyrosine kinases (TKs) is a family of tyrosine protein kinases with important functions in the regulation of a broad variety of physiological cell processes. Overactivity of TK disturbs cellular homeostasis and has been linked to the development of certain diseases, including various fibrotic diseases. In regard to liver fibrosis, several TKs, such as vascular endothelial growth factor receptor, platelet-derived growth factor receptor, fibroblast growth factor receptor, and epidermal growth factor receptor kinases, have been identified as central mediators in collagen production and potential targets for anti-liver fibrosis therapies. Given the essential role of TKs during liver fibrogenesis, multitargeted inhibitors of aberrant TK activity, including sorafenib, erlotinib, imatinib, sunitinib, nilotinib, brivanib and vatalanib, have been shown to have potential for treating liver fibrosis. Beneficial effects are observed by researchers of this field using these multitargeted TK inhibitors in preclinical animal models and in patients with liver fibrosis. The present review will briefly summarize the anti-liver fibrosis effects of multitargeted TK inhibitors and molecular mechanisms.