Arsenic trioxide augments immunogenic cell death and induces cGAS-STING-IFN pathway activation in hepatocellular carcinoma.

The treatment of hepatocellular carcinoma (HCC) is particularly challenging due to the inherent tumoral heterogeneity and easy resistance towards chemotherapy and immunotherapy. Arsenic trioxide (ATO) has emerged as a cytotoxic agent effective for treating solid tumors, including advanced HCC. However, its effectiveness in HCC treatment remains limited, and the underlying mechanisms are still uncertain. Therefore, this study aimed to characterize the effects and mechanisms of ATO in HCC. By evaluating the susceptibilities of human and murine HCC cell lines to ATO treatment, we discovered that HCC cells exhibited a range of sensitivity to ATO treatment, highlighting their inherent heterogeneity. A gene signature comprising 265 genes was identified to distinguish ATO-sensitive from ATO-insensitive cells. According to this signature, HCC patients have also been classified and exhibited differential features of ATO response. Our results showed that ATO treatment induced reactive oxygen species (ROS) accumulation and the activation of multiple cell death modalities, including necroptosis and ferroptosis, in ATO-sensitive HCC cells. Meanwhile, elevated tumoral immunogenicity was also observed in ATO-sensitive HCC cells. Similar effects were not observed in ATO-insensitive cells. We reported that ATO treatment induced mitochondrial injury and mtDNA release into the cytoplasm in ATO-sensitive HCC tumors. This subsequently activated the cGAS-STING-IFN axis, facilitating CD8+ T cell infiltration and activation. However, we found that the IFN pathway also induced tumoral PD-L1 expression, potentially antagonizing ATO-mediated immune attack. Additional anti-PD1 therapy promoted the anti-tumor response of ATO in ATO-sensitive HCC tumors. In summary, our data indicate that heterogeneous ATO responses exist in HCC tumors, and ATO treatment significantly induces immunogenic cell death (ICD) and activates the tumor-derived mtDNA-STING-IFN axis. These findings may offer a new perspective on the clinical treatment of HCC and warrant further study.

Comparable effects of Jiedu Granule, a compound Chinese herbal medicine, and sorafenib for advanced hepatocellular carcinoma: A prospective multicenter cohort study.

OBJECTIVE: Sorafenib has been extensively used for the treatment of advanced hepatocellular carcinoma (HCC), and Chinese herbal medicine has also been used to manage advanced HCC. The present work evaluates the effectiveness and safety of Jiedu (JD) Granule, a compound of traditional Chinese herbal medicine, side-by-side with sorafenib for the treatment of advance HCC. METHODS: Patients with advanced HCC receiving treatment with JD Granule or sorafenib were enrolled from December 2014 to March 2018. The primary endpoint was overall survival (OS). The secondary endpoints were progression-free survival (PFS) and safety. Propensity score matching (PSM) analysis was used to control for possible selection bias from the study group allocation process. RESULTS: Of the 325 patients included, 161 received JD Granule and 164 received sorafenib. No significant differences were found in OS or PFS among patients receiving JD Granule compared to sorafenib (P > 0.05). Median OS of the two study groups was 6.83 months (95% confidence interval [CI]: 5.83-9.47) in the group receiving JD Granule and 8 months (95% CI: 6.67-9.80) in the group receiving sorafenib, with half-, 1- and 2-year survival rates of 53.6%, 31.2% and 13.2% vs 60.1%, 35.5% and 14.2%, respectively. Even after PSM, the median survival time did not differ between the JD Granule group (9.03 months; 95% CI: 6.37-14.2) and the sorafenib group (7.93 months; 95% CI: 6.5-9.97), with comparable half-, 1- and 2-year survival rates. The most common adverse events (AEs) were diarrhea (13.7%) and fatigue (5.6%) in the JD Granule group, and hand-foot skin reaction (46.3%) and diarrhea (36.6%) in the sorafenib group. The JD Granule was more cost-effective than sorafenib treatment for advanced HCC. CONCLUSION: Compared to sorafenib, JD Granule was more cost-effective and caused fewer AEs for the treatment of Chinese patients with advanced HCC.

Antimetastatic effects of norcantharidin on hepatocellular carcinoma cells by up-regulating FAM46C expression.

Hepatocellular carcinoma (HCC) is the third leading cause of cancer-related mortality worldwide. Norcantharidin (NCTD), a demethylated analog of cantharidin, possesses antimetastatic effects on HCC cells. The aim of this study was to identify target proteins of NCTD. In this study, we confirmed the antimetastatic effects of NCTD on SMMC-7721 and MHCC-97H cells. Through RNA sequencing, we found a non-canonical poly (A) polymerase, Family-with-sequence-similarity-46C (FAM46C) was up-regulated in response to NCTD exposure. Gene set enrichment analysis on The Cancer Genome Atlas liver HCC (LIHC) dataset revealed that metastasis down pathway was strongly associated with FAM46C expression. Overexpression of FAM46C in HCC cells suppressed cell migration and invasion via suppressing transforming growth factor-ß (TGF-ß)/Smad signaling and epithelial-mesenchymal transition (EMT) process. Additionally, the antimetastatic effects of NCTD on HCC cells were partially rescued by FAM46C knockdown. Collectively, our results suggested that FAM46C, up-regulated by NCTD treatment, played a critical role in promoting the migration and invasion of HCC cells via TGF-ß/Smad signaling. We identified a new therapeutic target of NCTD.

Nomograms for survival prediction in patients undergoing liver resection for hepatitis B virus related early stage hepatocellular carcinoma.

BACKGROUND: The long-term outcomes of patients who underwent liver resection (LR) for early-stage hepatitis B virus (HBV)-related hepatocellular carcinomas (HCCs) are difficult to predict. This study aimed to develop two nomograms to predict postoperative disease-free survival (DFS) and overall survival (OS), respectively. METHODS: Data on a primary cohort of 1328 patients who underwent LR for HBV-related HCCs within Milan criteria at the Eastern Hepatobiliary Surgery Hospital (EHBH) from 2000 to 2006 were used to develop the nomograms by the Cox regression analyses. An internal validation cohort of 442 patients operated from 2006 to 2011 at the EHBH and an external validation cohort of 474 patients operated from 2007 to 2009 at the Zhongshan Hospital were used for validation studies. Discrimination and calibration were measured using concordance index (C-index), calibration plots and Kaplan-Meier curves. RESULTS: The independent predictors of DFS or OS which included tumour stage factors, biomarker and HBV-DNA level were respectively incorporated into the two nomograms. In the primary cohort, the C-indexes of the models in predicting DFS and OS were 0.76 (95% confidence interval: 0.75-0.78) and 0.79 (0.77-0.81), respectively. The calibration curves fitted well. Both nomograms accurately stratify patients into four distinct incremental prognostic subgroups. The C-indexes of the nomogram for OS prediction was significantly higher than those of the six conventional staging systems (0.65-0.71, all P<0.001). These results were verified by the internal and external validations. CONCLUSION: The proposed nomograms showed good prognostication for patients with early HBV-related HCCs after hepatectomy.

A novel scoring system predicts adjuvant chemolipiodolization benefit for hepatocellular carcinoma patients after hepatectomy.

Our aim in this study was to develop a prognostic scoring system with which to identify patients most likely to benefit from adjuvant chemolipiodolization (ACL) after liver resection for hepatocellular carcinoma (HCC). Data from 1150 HCC patients who underwent liver resection between 2002 and 2008 at the Eastern Hepatobiliary Surgery Hospital were used to develop the scoring system. Patients were stratified into prognostic subgroups using the new scoring system, and the outcomes of patients who received ACL and those who did not were compared in each subgroup. Using data from 379 patients operated on between 2008 and 2010 for validation, the scoring system had a concordance index (C-index) of 0.75 for predicting post-resectional overall survival (OS). It optimally stratified patients into three prognostic subgroups with scores of 0-5, 6-9 and ≥ 10, having better, medium and worse survival outcomes, respectively. A difference in OS between ACL and non-ACL patients was only detected in the subgroup with scores ≥ 10 (1-, 3-, and 5-year OS rates: 63.9%, 22.6%, and 9.0% vs. 33.8%, 5.6%, and 2.8%, p = 0.001). Our proposed scoring system provides an effective tool for selecting the patients most likely to benefit from ACL.

Loss of function of Notch1 identifies a poor prognosis group of early stage hepatocellular carcinoma following hepatectomy.

Notch1 has previously been implicated in the carcinogenesis of hepatocellular carcinoma (HCC). The present study aimed to investigate the prognostic value of Notch1 in early stage HCC patients after hepatectomy. The differential expression of Notch1 in paired tumor and non-tumorous tissue was evaluated by RT-PCR, western blotting and immunohistochemistry. The correlation between Notch1 expression and the surgical outcome of patients at BCLC stage 0/A and its ≤5 cm subgroup was retrospectively investigated in 206 patients from the Eastern Hepatobiliary Surgery Hospital (training cohort), and prospectively validated in 185 patients from the same center and retrospectively verified in 129 patients from the Fujian Medical University (validation cohort 1 and 2, respectively). Compared with paired non-tumorous tissues, loss of Notch1 was observed in tumor tissue. Patients with normal Notch1 had better prognosis than those with loss of Notch1 in the training cohort and ≤5 cm subgroup (time to recurrence: 38.5±6.1 vs. 16.0±3.2 months, P<0.001 and 53.0±6.1 vs. 21.7±3.5 months, P=0.004; 1-, 3-, 5-year survival rates: 91, 64 and 49% vs. 73, 31 and 22%, P<0.001 and 93, 71, 57% vs. 76, 39, 24%, P<0.001). Notch1 expression was an independent factor for recurrence and survival (hazard ratio: 1.901, 2.154; 2.038 and 2.337). Moreover, Notch1 status affected early tumor recurrence, as the 2-year recurrence rate was 61.2 vs. 26.9% (P<0.001) and 51.2 vs. 21.3% (P=0.002) in tumors with reduced or increased Notch1 expression in this cohort and subgroup. These results were fully confirmed by the study in our prospective and retrospective validation cohorts. The status of Notch1 is useful for predicting the prognosis of patients with early stage HCC undergoing hepatectomy.

Serum thioredoxin is a diagnostic marker for hepatocellular carcinoma.

Here we found that serum levels of thioredoxin were increased in patients with hepatocellular carcinoma (HCC). The optimum diagnostic cutoff for thioredoxin was 20.5 ng/mL (area under curve [AUC] 0.946 [95% CI 0.923-0.969] in the training cohort; 0.941 [0.918-0.963] in the validation cohort). High serum concentrations of thioredoxin differentiated HCC from chronic liver diseases and cirrhosis (0.901 [0.875-0.923] in the training cohort; 0.906 [0.870-0.925] in the validation cohort). Furthermore, a higher proportion of patients with very early HCC had positive results for thioredoxin than for alpha-Fetoprotein (AFP) (73.7% VS.31.6%; P < 0.0001). Among AFP-negative patients with very early HCC, 18 (69.2%) of 26 had positive thioredoxin results. Our results indicate that serum thioredoxin complements measurement of AFP in the diagnosis of HCC, especially in very early disease. Combined model (thioredoxin and AFP) showed a significantly greater discriminatory ability as compared with those markers alone.

Inhibitory effects of taurine and oat fiber on intestinal endotoxin release in rats.

Numerous studies have demonstrated that endotoxin plays an important role in the development and progression of hepatic cirrhosis. However, there is no effective remedy for the prevention and treatment of intestinal endotoxemia. Taurine has been reported to have beneficial effects on endotoxemia. Oats have been shown to absorb intestinal toxins and increase excretion of intestinal toxins. The present study was to investigate whether a combination of taurine and oat has an additive inhibitory effect on endotoxin release in a rat liver ischemia/reperfusion model. Our results showed that the combination of taurine (300mgkg(-1)d(-1)) and oat fiber (15gkg(-1)d(-1)) significantly reduced endotoxin levels in the portal vein by 36.3% when compared to the control group (0.168+/-0.035Eu/ml in the treatment group vs 0.264+/-0.058Eu/ml in the control group, P<0.01). The treatment of taurine (300mgkg(-1)d(-1)) and oat fiber (15gkg(-1)d(-1)) induced 21.5% and 18.4% reduction in endotoxin levels, respectively, when compared to the control group (P<0.05). We conclude that the combination of taurine and oat fiber achieved an additive inhibitory effect on intestinal endotoxin release, which might be an effective approach for the treatment of intestinal endotoxemia.

Hepatoprotective and anti-hepatocarcinogenic effects of glycyrrhizin and matrine.

Matrine (Mat), a component extracted from Sophora flavescens Ait, has a wide spectrum of pharmacological effects. Glycyrrhizin (Gly), a major active constituent of licorice (Glycyrrhiza glabra) root, has various pharmacological effects. Gly and Mat are ancillary drugs used clinically in China for protection of liver function and treatment of tumors. However, habitual administration of Gly may cause adverse effects marked by the development of pseudohypercorticosteroidism. This work was designed to see whether combination use of Gly and Mat could offer better liver protective and anti-hepatocarcinogenic effects than Gly or Mat alone, and whether it could reduce the adverse effects of Gly alone by acetaminophen-induced hepatotoxicity, diethylnitrosamine-induced hepatocarcinogenesis, induction of immunosuppression, albumen-induced swelling of rat hind paws. The results showed that compared with Gly or Mat alone, Gly+Mat reduced the mortality of acetaminophen overdosed mice more effectively, attenuate acetaminophen-induced hepatotoxicity, and reduced the number and area of gamma-GT positive foci, thus protecting liver function and preventing HCC from occurring. In addition, Gly+Mat had a protective effect on immunosuppression, a strong non-specific anti-inflammatory effect, and an effect of reducing the incidence of sodium and water retention.

[Detection of estrogenic effects of nonylphenol and bisphenol A in vitro reporter gene-based assays].

OBJECTIVE: To explore the estrogenic effects and disruptive mechanism of NP and BPA by reporter gene-based assays we developed. METHODS: pERE-Luc plamid was generated by inserting estrogen response element (ERE) fragment into MCS of pGL3-promoter vector. MCF7 cells were cotransfected with pERE-Luc and phRL-SV40 using Sofast transfection reagent. The cells then treated with 17beta-estradiol (E2), tamoxifen (Tam), nonylphenol(NP) and bisphenol A (BPA) and expression of the repoter gene in the cell lysates was assayed using Dual-Lucferase reporter assay system. RESULTS: The pERE-Luc plasmid was constructed. Luciferase activities of MCF7 cells transfected pERE-Luc showed dose-responed realitionship with E2. 1 x 10(-11) mol/L E2 could induce the expression of reporter gene and 1 x 10(-9) mol/L E2 resulted in the largest luciferase activity. E2 couldn't induce the luciferase activity without pERE-Luc. Tam is a complete antagonist, inhibited the E2-induced luciferase expression. NP induced the luciferase activity at concertrations > 1 x 10(-6) mol/L, BPA induced the luciferase activity at concertrations > 1 x 10(-6) mol/L. The estrogenic activity of NP was more than BPA. CONCLUSION: The assay we established is usful, NP and BPA showed estrogenic activities.

[Clinical observation of cinobufacini injection used to treat moderate and advanced primary liver cancer].

OBJECTIVE: To observe the clinical effect of cinobufacini injection in treating moderate and advanced primary liver cancer (PLC). METHODS: One hundred patients with moderate and advanced PLC were randomly divided into cino-treated group (50 patients) and control group (50 patients). The quality of life, tumor size, some changes of laboratory tests, and survival time were observed. RESULTS: The progressive rate of cino-treated group (18%) was lower than that of the control group (32%). The quality of life of the cino-treated group (80%) was better than that of the control group (72%), but without statistical significance. The survival rate of >12 months of the cino-treated group (30%) was higher than that of the control group (18%). The patients' liver function such as serum total bilirubin and ALT decreased obviously in the cino-treated group while increased a lot in the control group. The level of AFP increased after treatment with statistical significance in the control group while there was no statistical significance in the cino-treated group. CONCLUSION: Cinobufacini injection can not only inhibit the proliferation of cancer, but also protect liver function, improve quality of life and prolong survival time.