Comparison of RETREAT and CCFSS Scoring Systems for Risk Estimation of Hepatocellular Carcinoma Recurrence After Radical Resection.

Objective: RETREAT and CCFSS are two scoring systems specifically designed to predict postoperative recurrence in hepatocellular carcinoma (HCC). RETREAT and CCFSS are scoring systems designed to predict postoperative recurrence in hepatocellular carcinoma (HCC). This study aims to evaluate and compare the prognostic performance of two scoring systems, RETREAT and CCFSS, for predicting tumor recurrence after radical resection in patients with hepatocellular carcinoma (HCC). Methods: In this retrospective study, we analyzed the clinical data of 124 patients with HCC who underwent radical resection at our hospital between March 2017 and February 2020. The patients were categorized into a recurrence group (n = 41) and a non-recurrence group (n = 83) based on whether they experienced recurrence within 3 years of follow-up. Logistic regression analysis was conducted to compare the clinical characteristics between the two groups and identify factors associated with HCC recurrence. Additionally, we generated receiver operating characteristic (ROC) curves to evaluate the predictive value of the RETREAT and CCFSS scoring systems for predicting HCC recurrence after surgery. Results: Significant differences (P < .05) were found in the postoperative relapse-free survival time, HBsAg positivity, preoperative ALT≥ 40 U/L, preoperative Alb < 40 g/L, pericardial invasion, RETREAT score, and CCFSS score between the two groups. A multi-factor logistic regression analysis was performed between postoperative relapse in HCC patients and HBsAg positivity, preoperative ALT ≥ 40U/L, preoperative Alb<40g/L, pericardial invasion, RETREAT score, and CCFSS score grading as independent variables. The analysis revealed that HBsAg positivity (OR = 6.039, 95%CI: 2.994~12.182), preoperative ALT ≥ 40 U/L (OR = 3.738, 95%CI: 2.016-6.931), preoperative Alb< 40g/L (OR = 3.655, 95%CI: 2.083-6.415), envelope invasion (OR = 3.119, 95%CI: 1.922-5.061), RETREAT score (OR = 6.867, 95%CI: 3.204-14.719), and CCFSS score (OR = 3.228, 95%CI: 1.951-5.342) were significant factors influencing postoperative recurrence in HCC patients (P < .05). The RETREAT score had an area under the curve (AUC) of 0.853, with a sensitivity of 89.23% and a specificity of 93.87% (95%CI: 0.7567-0.950), while the AUC of the CCFSS score for predicting postoperative recurrence in HCC patients was 0.741, with a sensitivity of 78.57% and a specificity of 60.71% (95%CI: 0.609-0.873). Among the factors analyzed, including postoperative relapse-free survival time, HBsAg positivity, preoperative ALT≥ 40 U/L, preoperative Alb < 40 g/L, pericardial invasion, RETREAT score, and CCFSS score, statistically significant differences were observed between the recurrence and non-recurrence groups (P < .05). Conclusion: The RETREAT score demonstrates higher efficacy compared to the CCFSS score in predicting postoperative recurrence in hepatocellular carcinoma (HCC) patients and can serve as a valuable tool for the prognostic evaluation of HCC patients who undergo radical resection. The RETREAT score's superiority can be attributed to its higher area under the curve (AUC) and improved sensitivity and specificity, which allow for better discrimination between positive and negative cases. This enhanced accuracy enables clinicians to make more informed decisions towards post-surgical management and treatment strategies, ultimately improving patient outcomes. Thus, the RETREAT score serves as a vital tool for personalized and targeted care towards HCC patients undergoing radical resection.

TGF-ß1-Mediated PD-L1 Glycosylation Contributes to Immune Escape via c-Jun/STT3A Pathway in Nasopharyngeal Carcinoma.

Immunotherapy targeting programmed death ligand-1/programmed cell death protein-1 (PD-L1/PD-1) has achieved great success in multiple cancers, but only a small subset of patients showed clinical responses. Recent evidences have shown that post-translational modification of PD-L1 protein could regulate its protein stability and interaction with cognate receptor PD-1, thereby affecting anticancer immunotherapy in several solid tumors. However, the molecular mechanisms underlying how PD-1/PD-L1 expression is regulated still remain unclear in nasopharyngeal carcinoma (NPC). Here, we found N-glycosylation of PD-L1 in NPC cells and tissues. Mechanistically, we showed that STT3A transferred N-linked glycans to PD-L1, and TGF-ß1 could positively regulate STT3A expression through activating c-Jun to bind to STT3A promoter. Functional assays showed that inhibition of TGF-ß1 resulted in a decrease of glycosylated PD-L1 and enhanced cytotoxic T-cell function against NPC cells. Analysis of clinical specimens revealed that the expression of STT3A was positively correlated with TGF-ß1 and c-Jun, and high STT3A expression was positively correlated with a more advanced clinical stage. Altogether, TGF-ß1 activated c-Jun/STT3A signaling pathway to promote N-glycosylation of PD-L1, thus further facilitating immune evasion and reducing the efficacy of cancer immunotherapy. As such, all these data suggested that targeting TGF-ß1 pathway might be a promising approach to enhance immune checkpoint blockade, and simultaneous blockade of PD-L1 and TGF-ß1 pathways might elicit potent and superior antitumor activity relative to monotherapies.

The effect of targeted regulation of LATS2 by LncRNA BCAR4 on proliferation, migration and apoptosis of HCC cells.

OBJECTIVE: This study explored and analyzed the effects of targeted regulation of LATS2 by LncRNA BCAR4 on the proliferation, migration and apoptosis of hepatocellular carcinoma (HCC). METHOD: We detected the expression of LncRNA, BCAR4 and LATS2 mRNA in liver hepatocellular carcinoma HepG2 cells and normal hepatocellular cells LO2 by RT-PCR. HepG2 cells were divided into BCAR4-siRNA, NC-siRNA and control groups. We detected the targeted regulation of LncRNA BCAR4 on LATS2 by luciferase gene assay, and measured the proliferation, migration and apoptosis of cells in each group by RT-PCR, MTT, Transwell and flow cytometry, respectively. RESULTS: The relative expression of LncRNA BCAR4 in HepG2 cells was critically higher than that in LO2 cells (P<0.05), while LATS2 mRNA in HepG2 cells was significantly less than that in LO2 cells (P<0.05). Compared with NC siRNA group, the content of luciferase in BCAR4 siRNA group was much higher (P<0.05); The relative expression of LncRNA BCAR4 in BCAR4 siRNA group decreased dramatically than that in NC-siRNA and control group (P<0.05), and the relative expression of LATS2 mRNA increased remarkably than that in NC-siRNA group and control group (P<0.05). The OD value of BCAR4 siRNA group was dramatically higher than that of NC-siRNA group and control group after 48 h and 72 h culture (P<0.05). The quantity of invaded cells in BCAR4 siRNA group was markedly less than that in NC-SIRNA group and control group (P<0.05). Cell apoptosis rate in BCAR4-siRNA group was significantly higher than that of NC-siRNA group and control group (P<0.05). CONCLUSION: LncRNA BCAR4 can regulate the LATS2 expression, and inhibiting the expression of LncRNA BCAR4 can inhibit proliferation, invasion of HepG2 cells and induce its apoptosis, which finding provides a certain reference for the targeted therapy of liver cancer.

Survival benefit of induction chemotherapy for locally advanced nasopharyngeal carcinoma: prognosis based on a new risk estimation model.

BACKGROUND: Although the National Comprehensive Cancer Network (NCCN) Guidelines recommend CCRT+AC and IC + CCRT as level 2A evidence for treatment of the locoregionally advanced NPC (II-IVa), IC + CCRT+AC could also be an alternative but it is seldom used because of the low completion rates. This article aimed to compare the effectiveness of the three radiotherapy regimens using a large-scale retrospective study. METHODS: This retrospective single center analysis enrolled 1812 diagnosed NPC patients at Nanfang Hospital from January 2005 to December 2015 and only 729 patients met the inclusion criteria and were analyzed. Patients without distant metastasis, age of 18-70 years, Karnofsky scores of at least 70,stage III-IVb, and adequate adequate bone marrow, liver and renal function. Were enrolled. Adverse events and other categorical variables were compared by Pearson chi-square test or Fishier exact test. Time-to-event data were described with the Kaplan-Meier curves, time-to-event intervals compared with the log-rank test. We did multivariable analyses with the Cox proportional hazards model to test the independent signifi cance of diff erent factors. Cox proportional hazards model was used to estimate the ß regression coeffi cient, p value, and hazard ratio and its 95% CI for each of the selected risk predictors. RESULTS: The median follow-up time was 47 months. Kaplan-Meier analyses revealed no significant differences among three groups in 3-year failure-free survival (FFS, P = 0.225), 3-year overall survival (OS, P = 0.992), 3-year locoregional failure-free survival (LFFS, P = 0.549), and 3-year distant failure-free survival (DFFS, P = 0.174). Stratified survival analysis based on the risk scoring model revealed no differences in FFS, OS, LFFS, and DFFS between IC + CCRT and CCRT+AC groups for low-risk patients, however, the 3-year OS (88.3% vs. 77.6%, P = 0.049) and 3-year DFFS (84.0% vs.66.8%, P = 0.032) were respectively significantly better in IC + CCRT group compared with CCRT+AC group for high-risk patients. CONCLUSIONS: Compared with CCRT+AC, IC + CCRT lowers distant metastasis rate and improves OS among patients with locally advanced NPC in high risk group.

Foxq1 promotes metastasis of nasopharyngeal carcinoma by inducing vasculogenic mimicry via the EGFR signaling pathway.

In nasopharyngeal carcinoma (NPC), the treatment of tumor metastasis and recurrence is challenging and is associated with poor clinical efficacy. Vasculogenic mimicry (VM) is a new blood-supply model of malignant tumor that is closely related to tumors' distant metastasis. Our previous study demonstrated that miR-124 could target Foxq1 to inhibit NPC metastasis. Whether Foxq1 affects metastasis through vasculogenic mimicry is worth consideration. In this study, we show that VM formation positively correlates with the expression of Foxq1, and EGFR, and the TNM stage in 114 NPC patient samples. Meanwhile, we show that VM-positive NPC patients have a poor prognosis. Furthermore, using in vitro and vivo approaches, we confirm that Foxq1 has a significant effect on NPC metastasis through promoting VM formation, which could be effectively inhibited by EGFR inhibitors (Nimotuzumab or Erlotinib). Also a synergistic efficacy of anti-EGFR and anti-VEGF drugs has been found in NPC inhibition. Mechanistically, the luciferase reporter gene and CHIP assays show that Foxq1 directly binds to the EGFR promoter region and regulates EGFR transcription. In conclusion, our results show that Foxq1 is regulated by miR-124 and that it promotes NPC metastasis by inducing VM via the EGFR signaling pathway. Overall, these results provide a new theoretical support and a novel target selection for anti-VM therapy in the treatment of nasopharyngeal carcinoma.

EBV-EBNA1 constructs an immunosuppressive microenvironment for nasopharyngeal carcinoma by promoting the chemoattraction of Treg cells.

BACKGROUND: Nasopharyngeal carcinoma (NPC) is primarily caused by the Epstein-Barr virus (EBV) infection in NPC endemic areas. EBNA1 is an EBV-encoded nuclear antigen, which plays a critical role in the maintenance and replication of EBV genome. However, the mechanisms of EBNA1-promoted NPC immune escape are unknown. Regulatory T (Treg) cells are among the key regulators in restraining antitumor responses. However, the mechanisms of accumulation of Treg cells in NPC have not been defined. This study attempted to identify the detailed mechanisms of EBNA1 functions as a tumor accelerator to promote NPC immune escape by enhancing chemoattraction of Treg cells. METHODS: mRNA profiles were determined by next-generation sequencing in NPC cells. In vitro and in vivo assays were performed to analyze the role of EBNA1 in regulation of recruitment of Treg cells. Colocation and coimmunoprecipitation analyzes were used to identify the SMAD3/c-JUN complex. Chromatin immunoprecipitation assay and dual luciferase reporter assays were designed to demonstrate c-JUN binding to miR-200a promoter and miR-200a targeting to CXCL12 3'Untranslated Regions. The hepatocellular carcinoma models were designed to demonstrate universality of the CXCL12-CXCR4-Treg axis in promoting immune evasion of various tumors. RESULT: A novel molecular mechanism was identified that involves EBV-EBNA1-stimulated chemotactic migration of Treg cells toward NPC microenvironment by upregulation of the transforming growth factor-ß1 (TGFß1)-SMAD3-PI3K-AKT-c-JUN-CXCL12-CXCR4 axis and downregulation of miR-200a. EBV-EBNA1 promotes the chemoattraction of Treg cells by governing the protein-protein interactions of the SMAD3/c-JUN complex in a TGFß1-dependent manner in vitro and in vivo. TGFß1 suppresses miR-200a by enhancing the SMAD3/c-JUN complex. miR-200a negatively regulates the CXCL12 chemokine by direct targeting of the CXCL12 3'UTR region. However, CXCL12 acts as the target gene of miR-200a and as an inhibitor of miR-200a transcription, and inhibition of miR-200a by CXCL12 is mediated by c-JUN, which directly binds to the miR-200a promoter and forms a c-JUN-miR-200a-CXCL12-c-JUN feedback loop. In addition, enhanced CXCL12 efficiently attracts CXCR4-positive Treg cells to remodel an immunosuppressive microenvironment. CONCLUSIONS: EBV-EBNA1 promotes chemotactic migration of Treg cells via the TGFß1-SMAD3-PI3K-AKT-c-JUN-miR-200a-CXCL12-CXCR4 axis in the NPC microenvironment. These results suggest that EBV-EBNA1 may serve as a potential therapeutic target to reshape the NPC microenvironment.

Mechanisms of Epstein-Barr virus nuclear antigen 1 favor Tregs accumulation in nasopharyngeal carcinoma.

BACKGROUND: Documented reports proved that Epstein-Barr virus (EBV) infection increased infiltration of Tregs in malignancy. However, the mechanism of EBV recruitment Tregs into nasopharyngeal carcinoma (NPC) tissues has not been detailed discussion. METHODS: Expression of EBV nuclear antigen 1 (EBNA1) and Foxp3 in NPC tissue samples was detected by immunohistochemistry. EBNA1+ NPC cell lines were used to coculture with PBMC, naïve T cells, Tregs, and monocytes. Percent of Treg was detected by flow cytometry. RESULTS: EBNA1 protein was overexpressed in NPC tissues, and was associated with a number of infiltrated Tregs. EBNA1+ NPC cells converted naïve T cells into Tregs by up-regulated TGF-ß1. Enhanced CCL20 production in EBNA1-expressed tumor cells increased Tregs migration. Polarized-M2 macrophages by EBNA1 expression cells converted naïve T cells into Tregs. CONCLUSIONS: EBNA1 favors accumulation of Tregs in NPC through: (a) upregulated TGF-ß1 converted naïve T cell into Treg; (b) upregulated CCL20 increased Treg migration; and (c) polarized-M2 macrophage converted naïve T cell into Treg.

Retrospective study comparing anti-EGFR monoclonal antibody plus cisplatin-based chemoradiotherapy versus chemoradiotherapy alone for stage II-IVb nasopharyngeal carcinoma and prognostic value of EGFR and VEGF expression.

OBJECTIVES: The benefits of adding anti-EGFR therapy to conventional chemoradiotherapy (CRT) for nasopharyngeal carcinoma (NPC) remain uncertain, possibly because only a subgroup of patients show better outcome. To address this issue, we compared the efficacy of CRT plus cetuximab (CTX) or nimotuzumab (NTZ) to CRT alone for stage II-IVb NPC and examined possible prognostic indicators, including tumour EGFR and VEGR expression levels. DESIGN, SETTING AND PARTICIPANTS: This retrospective study enrolled 1812 patients at initial NPC diagnosis at Nanfang Hospital Affiliated to Southern Medical University between January 2005 and December 2015. The cetuximab or nimotuzumab plus CRT group (CRT+NTZ/CTX) and the conventional chemoradiotherapy group (CRT) were matched by propensity scoring at 1:5, yielding 282 patients at clinical stage II-IVb with 47 in the CRT+NTZ/CTX group and 235 in the CRT group. MAIN OUTCOME MEASURES: The endpoints of the present study were locoregional relapse-free survival (LRFS), distant metastasis-free survival (DMFS), overall survival (OS). Immunohistochemistry (IHC) was used to investigate EGFR and VEGF expression levels in 31 patients of the CRT+NTZ/CTX group. RESULTS: There were no significant differences in LRFS, DMFS and OS, haematologic toxicity reactions, and gastrointestinal reactions between CRT+NTZ/CTX and CRT groups. There was a positive correlation between EGFR and VEGF expression levels. Among CRT+NTZ/CTX patients, those with high EGFR and VEGF expression levels exhibited better DMFS. CONCLUSIONS: Addition of anti-EGFR to cisplatin-based CRT appears to benefit only a subset of stage II-IVb NPC patients, those with elevated EGFR and VEGF expression levels.

Tumor cells induced-M2 macrophage favors accumulation of Treg in nasopharyngeal carcinoma.

Tumor-associated macrophage (TAM) is major component of tumor immune related inflammation and plays a crucial role in tumor immune escape. However, there have been no studies reported the relationship of TAM and immunosuppressive cell regulatory T cell (Treg) in NPC. This study is to discuss the interaction of TAM and Treg in NPC. In the present study, immunopathological assays demonstrated that M2 macrophage increased in NPC tissues. M2 macrophage had a positive correlation with aberrantly increased Foxp3+ Treg in NPC tissues. High density M2 macrophage and high density Treg predicted poor survival of NPC patients. In vitro studies using tumor cells co-cultured with monocyte indicated that tumor cells could induce monoctye into M2 macrophage via TGF-ß1 and IL-10. Tumor cells induced-M2 macrophage could induce the chemotaxis of Treg and increased the percent of Treg in PBMC. Our results proved that NPC cells induced M2 macrophage via TGF-ß1 and IL-10, and tumor cells induced-M2 macrophage could favor accumulation of Treg by conversion and chemotaxis.