PI3K/AKT phosphorylation activates ERRα by upregulating PGC­1α and PGC­1ß in gallbladder cancer.

The nuclear estrogen­related receptor­α (ERRα) is an orphan receptor that has been identified as a transcriptional factor. Peroxisome proliferator­activated receptor­Î³ (PPARγ) coactivator­1­α (PGC­1α) and PPARγ coactivator­1­ß (PGC­1ß) act as the co­activators of ERRα. Our previous study reported that activated ERRα promoted the invasion and proliferation of gallbladder cancer cells by promoting PI3K/AKT phosphorylation. Therefore, the aim of the current study was to investigate whether PI3K/AKT phosphorylation could enhance ERRα activity in a positive feedback loop. LY294002 and insulin­like growth factor I (IGF­I) were used to inhibit and promote PI3K/AKT phosphorylation, respectively. A 3X ERE­TATA luciferase reporter was used to measure ERRα activity. The present study found that LY294002 inhibited PI3K/AKT phosphorylation, decreased the proliferation and invasion of NOZ cells and suppressed the activity of ERRα. Conversely, IGF­I induced PI3K/AKT phosphorylation, promoted the proliferation and invasion of NOZ cells and enhanced the activity of ERRα. The protein expression levels of PGC­1α and PGC­1ß were elevated and reduced by IGF­I and LY294002, respectively. Moreover, knockdown of PGC­1α and PGC­1ß antagonized ERRα activation, which was enhanced by PI3K/AKT phosphorylation. Taken together, the present study demonstrated that PI3K/AKT phosphorylation triggered ERRα by upregulating the expression levels of PGC­1α and PGC­1ß in NOZ cells.

Prognostic significance of regional lymphadenectomy in T1b gallbladder cancer: Results from 24 hospitals in China.

BACKGROUND: Whether regional lymphadenectomy (RL) should be routinely performed in patients with T1b gallbladder cancer (GBC) remains a subject of debate. AIM: To investigate whether RL can improve the prognosis of patients with T1b GBC. METHODS: We studied a multicenter cohort of patients with T1b GBC who underwent surgery between 2008 and 2016 at 24 hospitals in 13 provinces in China. The log-rank test and Cox proportional hazards model were used to compare the overall survival (OS) of patients who underwent cholecystectomy (Ch) + RL and those who underwent Ch only. To investigate whether combined hepatectomy (Hep) improved OS in T1b patients, we studied patients who underwent Ch + RL to compare the OS of patients who underwent combined Hep and patients who did not. RESULTS: Of the 121 patients (aged 61.9 ± 10.1 years), 77 (63.6%) underwent Ch + RL, and 44 (36.4%) underwent Ch only. Seven (9.1%) patients in the Ch + RL group had lymph node metastasis. The 5-year OS rate was significantly higher in the Ch + RL group than in the Ch group (76.3% vs 56.8%, P = 0.036). Multivariate analysis showed that Ch + RL was significantly associated with improved OS (hazard ratio: 0.51; 95% confidence interval: 0.26-0.99). Among the 77 patients who underwent Ch + RL, no survival improvement was found in patients who underwent combined Hep (5-year OS rate: 79.5% for combined Hep and 76.1% for no Hep; P = 0.50). CONCLUSION: T1b GBC patients who underwent Ch + RL had a better prognosis than those who underwent Ch. Hep + Ch showed no improvement in prognosis in T1b GBC patients. Although recommended by both the National Comprehensive Cancer Network and Chinese Medical Association guidelines, RL was only performed in 63.6% of T1b GBC patients. Routine Ch + RL should be advised in T1b GBC.

Protocol for a gallbladder cancer registry study in China: the Chinese Research Group of Gallbladder Cancer (CRGGC) study.

INTRODUCTION: Gallbladder cancer (GBC), the sixth most common gastrointestinal tract cancer, poses a significant disease burden in China. However, no national representative data are available on the clinical characteristics, treatment and prognosis of GBC in the Chinese population. METHODS AND ANALYSIS: The Chinese Research Group of Gallbladder Cancer (CRGGC) study is a multicentre retrospective registry cohort study. Clinically diagnosed patient with GBC will be identified from 1 January 2008 to December, 2019, by reviewing the electronic medical records from 76 tertiary and secondary hospitals across 28 provinces in China. Patients with pathological and radiological diagnoses of malignancy, including cancer in situ, from the gallbladder and cystic duct are eligible, according to the National Comprehensive Cancer Network 2019 guidelines. Patients will be excluded if GBC is the secondary diagnosis in the discharge summary. The demographic characteristics, medical history, physical examination results, surgery information, pathological data, laboratory examination results and radiology reports will be collected in a standardised case report form. By May 2021, approximately 6000 patient with GBC will be included. The clinical follow-up data will be updated until 5 years after the last admission for GBC of each patient. The study aimed (1) to depict the clinical characteristics, including demographics, pathology, treatment and prognosis of patient with GBC in China; (2) to evaluate the adherence to clinical guidelines of GBC and (3) to improve clinical practice for diagnosing and treating GBC and provide references for policy-makers. ETHICS AND DISSEMINATION: The protocol of the CRGGC has been approved by the Committee for Ethics of Xinhua Hospital, Shanghai Jiao Tong University School of Medicine (SHEC-C-2019-085). All results of this study will be published in peer-reviewed journals and presented at relevant conferences. TRIAL REGISTRATION NUMBER: NCT04140552, Pre-results.

Prognostic value of precise hepatic pedicle dissection in anatomical resection for patients with hepatocellular carcinoma.

The present study aimed to investigate the long-term and perioperative outcomes of precise hepatic pedicle dissection in anatomical resection (precise AR) vs non-anatomical resection (NAR) for hepatocellular carcinoma (HCC) patients.Data from a total of 270 consecutive HCC patients who underwent curative hepatectomy were retrospectively collected. Propensity score matching (PSM) analysis was performed. The long-term outcomes of precise AR and NAR were analyzed using the Kaplan-Meier method and the Cox proportional hazards model.The 1-, 3-, and 5-year overall survival (OS) rates were 90.3%, 76.2%, and 65.7% in the PS-precise AR group, respectively (n = 103); and 88.3%, 70.5%, and 52.0% in the PS-NAR group, respectively (n = 103) (P = .043). The 1-, 3-, and 5-year recurrence-free survival (RFS) rates were 83.4%, 63.2%, and 46.0% in the PS-precise AR group, respectively; and 75.7%, 47.4%, and 28.3% in the PS-NAR group, respectively (P = .002). Multivariate analysis showed that ICG-R15, BCLC staging, and microvascular invasion (MVI) were independent risk factors for OS; while tumor size, types of resection, surgical margin, and MVI were independent risk factors for RFS. Subgroup analysis indicated that the RFS rate was significantly better in the PS-precise AR group than in the PS-NAR group for patients with MVI and tumor size ≤5 cm.After PSM, precise hepatic pedicle dissection in AR significantly improved the recurrence-free survival rate of solitary HCC patients compared with NAR, especially in those with MVI and tumor size ≤5 cm.

Estrogen-related receptor-α promotes gallbladder cancer development by enhancing the transcription of Nectin-4.

Estrogen-related receptor-α (ERRα) is a nuclear receptor of transcription factor that binds to estrogen responsive elements and estrogen-related responsive elements. Estrogen-related receptor-α is involved in metabolic processes and implicated in the progression and growth of several human malignancies. However, the biologic role and clinical significance of ERRα in gallbladder cancer (GBC) remains to be clarified. Here, we reported that ERRα protein expression was notably higher in GBC tissues than in cholecystitis tissues, and that the aberrantly higher ERRα expression was positively correlated with advanced TNM stage and indicated dismal prognosis of GBC (P < .01). In GBC cell lines NOZ and OCUG, the targeted depletion of ERRα retarded the growth and suppressed the migration and invasive capabilities of GBC cells, and inhibited epithelial-mesenchymal transition by decreasing the expression of mesenchymal markers and elevating the expression of epithelial markers. Moreover, ERRα knockdown inhibited tumor growth in nude mice and led to decreased expression levels of Nectin-4, p-PI3K p85α, and p-AKT. Overexpression of ERRα in the GBC-SD cell line showed exactly the opposite effect. The targeted inhibition of Nectin-4 antagonized GBC cell proliferation and invasion, which were induced by ERRα upregulation. Moreover, Nectin-4 depletion inhibited the ERRα-induced activation of the PI3K/AKT pathway. Chromatin immunoprecipitation analysis and dual-luciferase reporter gene assays showed that ERRα enhanced the transcription of Nectin-4 by binding to the promoter of Nectin-4. In conclusion, our data indicated that ERRα could be a potential target for the genetic treatment of GBC.

Anti-IL-20 monoclonal antibody suppresses hepatocellular carcinoma progression.

Interleukin (IL)-20 is a member of the IL-10 family of cytokines, which has been reported to participate in autoimmune inflammatory diseases. However, the potential role of IL-20 in hepatocellular carcinoma (HCC) progression has not yet been investigated. In the present study, it was observed that IL-20 mRNA and protein levels were markedly increased in the HCC tissues examined via reverse transcription-quantitative polymerase chain reaction and immunohistochemical staining. In addition, IL-20 expression was significantly associated with tumor size, metastasis, TNM stage and poor prognosis in patients with HCC. Mouse recombinant IL-20 (mIL-20) enhanced liver cancer cell proliferation, migration and invasion in vitro, while the anti-IL-20 monoclonal antibody (mAb) attenuated the effect of mIL-20, inhibiting cancer cell migration and invasion in vitro and suppressing cell growth in vitro and in vivo. This was detected by Cell Counting Kit-8, colony formation, Transwell assays and a xenograft tumor nude mouse model. Western blotting revealed that IL-20 promoted HCC progression through inducing transforming growth factor-ß and matrix metalloproteinase 9 expression and enhancing the phosphorylation of Jun N-terminal kinase and signal transducer and activator of transcription 3. The results of the present study indicated that IL-20 promotes HCC development. In addition, anti-IL-20 mAb may attenuate the effect of IL-20 and suppress liver tumorigenesis in vitro and in vivo, indicating that anti-IL-20 mAbs may potentially serve as effective therapeutic agents for HCC.

Prognostic value and preoperative predictors of microvascular invasion in solitary hepatocellular carcinoma ≤ 5 cm without macrovascular invasion.

OBJECTIVES: The aim of this study was to investigate the prognostic value and preoperative predictors of microvascular invasion (MVI) in solitary hepatocellular carcinoma (HCC) ≤ 5 cm without macrovascular invasion. METHODS: A total of 233 consecutive HCC patients underwent curative hepatectomy were included in our study. Independent risk factors influencing the prognosis were identified, and preoperative predictors for MVI were determined. RESULTS: Multivariate regression analysis identified ICG-R15, BCLC staging and MVI as independent risk factors for the overall survival rate. Type of resection and MVI were independent risk factors for the recurrence-free survival rate. Kaplan-Meier analysis showed the overall survival and recurrence-free survival rates in patients with MVI were significantly poorer than those in patients without MVI (P = 0.002 and P = 0.001). Anatomical resection obviously improved the overall survival and recurrence-free survival rates in patients with MVI compared with non-anatomical resection (P = 0.017 and P = 0.009). A prediction scoring system for MVI was built up according to the three independent predictors (tumor size > 3.5 cm, AFP > 200 ng/mL and GGT > 53 U/L). The prevalence of MVI in HCC patients with predictive score ≥ 2 was 58.3%, which was obviously higher than patients with predictive score < 2 (20.8%). CONCLUSIONS: MVI is associated with a poor prognosis in solitary HCC ≤ 5 cm after hepatectomy. Anatomical resection could improve the prognosis of HCC patients with MVI. The preoperative prediction scoring model has practical value for the prediction of MVI.

Development and validation of a novel predictive scoring model for microvascular invasion in patients with hepatocellular carcinoma.

PURPOSE: Microvascular invasion (MVI) in patients with hepatocellular carcinoma (HCC) cannot be accurately predicted preoperatively. This study aimed to establish a predictive scoring model of MVI in solitary HCC patients without macroscopic vascular invasion. METHODS: A total of 309 consecutive HCC patients who underwent curative hepatectomy were divided into the derivation (n=206) and validation cohort (n=103). A predictive scoring model of MVI was established according to the valuable predictors in the derivation cohort based on multivariate logistic regression analysis. The performance of the predictive model was evaluated in the derivation and validation cohorts. RESULTS: Preoperative imaging features on CECT, such as intratumoral arteries, non-nodular type of HCC and absence of radiological tumor capsule were independent predictors for MVI. The predictive scoring model was established according to the ß coefficients of the 3 predictors. Area under receiver operating characteristic (AUROC) of the predictive scoring model was 0.872 (95% CI, 0.817-0.928) and 0.856 (95% CI, 0.771-0.940) in the derivation and validation cohorts. The positive and negative predictive values were 76.5% and 88.0% in the derivation cohort and 74.4% and 88.3% in the validation cohort. The performance of the model was similar between the patients with tumor size ≤5cm and >5cm in AUROC (P=0.910). CONCLUSIONS: The predictive scoring model based on intratumoral arteries, non-nodular type of HCC, and absence of the radiological tumor capsule on preoperative CECT is of great value in the prediction of MVI regardless of tumor size.

Identification of miR-601 as a novel regulator in the development of pancreatic cancer.

Pancreatic cancer (PC) is one of the most aggressive malignancies, with a high mortality. Distant metastasis and recurrence are the main causes of PC-related deaths. MicroRNAs (miRNAs) have been reported in the serum or tumor tissue of cancer patients, including PC, which makes them potential biomarkers. The dysfunction of many miRNAs has been linked to PC occurrence and metastasis. In the current study, we found that miR-601 expression was significantly lower in PC samples, especially in metastatic compared to non-metastatic PC tissues. Gain-of-function and loss-of-function analysis showed that miR-601 suppressed PC cell proliferation and migration. One potential mechanism is that miR-601 inhibited the expression of Sirtuin 1 (SIRT1), which is a well-known regulator of PC development. We found that overexpression of SIRT1 could reverse the effect of miR-601 on PC cells. Our investigation therefore suggests that both miR-601 and SIRT1 are possible biomarkers for the early detection, and targets for the treatment of PC.

Prognostic value of a novel risk classification of microvascular invasion in patients with hepatocellular carcinoma after resection.

OBJECTIVES: The present research aimed to evaluate the prognostic value of a novel risk classification of microvascular invasion (MVI) in hepatocellular carcinoma (HCC) after resection. METHODS: A total of 295 consecutive HCC patients underwent hepatectomy were included in our study. We evaluated the degree of MVI according to the following three features: the number of invaded microvessels (≤5 vs >5), the number of invading carcinoma cells (≤ 50 vs >50), the distance of invasion from tumor edge (≤1 cm vs >1 cm). RESULTS: All patients were divided into three groups according to the three risk factors of MVI: non-MVI group (n=180), low-MVI group (n=60) and high-MVI group (n=55). The overall survival (OS) and recurrence-free survival (RFS) rates of high-MVI group were significantly poorer than those of low-MVI and non-MVI groups (P<0.001 and P=0.001; P<0.001 and P=0.003). Multivariate analysis showed high-MVI, type of resection, ICG-R15 and tumor size were risk factors for OS after hepatectomy. High-MVI, type of resection and tumor size were risk factors for RFS. In subgroup analyses, the OS and RFS rates of low-MVI and non-MVI groups were better than high-MVI group regardless of tumor size. In high-MVI group, anatomical liver resection (n=28) showed better OS and RFS rates compared with non-anatomical liver resection (n=29) (P=0.012 and P=0.002). CONCLUSIONS: The novel risk classification of MVI based on histopathological features is valuable for predicting prognosis of HCC patients after hepatectomy.

Preoperative Evaluation of Malignant Perihilar Biliary Obstruction: Negative-Contrast CT Cholangiopancreatography and CT Angiography Versus MRCP and MR Angiography.

OBJECTIVE: The purpose of this study was to compare negative-contrast CT cholangiopancreatography (CTCP) and CT angiography (CTA) with MRCP and MR angiography (MRA) for the preoperative evaluation of malignant perihilar biliary obstruction. MATERIALS AND METHODS: Twenty-one patients with pathologically proven malignant perihilar biliary obstructions who had undergone both CT and MRI examinations were reviewed retrospectively. Two reviewers independently analyzed the two image sets-the negative-contrast CTCP and CTA images (i.e., CT set) and the MRCP and MRA images (i.e., MRI set)-in preoperatively evaluating the classification of malignant perihilar biliary obstruction, hepatic artery and portal vein invasion, nodal metastasis, and organ spread. The results were compared with surgical and pathologic records. RESULTS: For the classification of malignant perihilar biliary obstruction on the two image sets, the accuracy was not statistically significant (p = 1.000 for reviewer 1 and p = 0.500 for reviewer 2). For the evaluation of portal vein invasion, nodal metastasis, and organ spread, the accuracies were also not statistically significantly different (p = 0.335, 0.339, and 0.781 for reviewer 1; and p = 0.403, 0.495, and 0.325 for reviewer 2, respectively). In the assessment of hepatic artery status, the accuracy was statistically significant (p = 0.046 for reviewer 1 and p = 0.036 for reviewer 2). CONCLUSION: Compared with the MRI set, the CT set provides equivalent performance in assessing the classification of malignant perihilar biliary obstruction, portal vein involvement, nodal metastasis, and organ spread, but has higher accuracy in assessing arterial invasion.

Meta-analysis of the cystatin C(CST3) gene G73A polymorphism and susceptibility to Alzheimer's disease.

BACKGROUND: The association between cystatin C(CST3) gene (rs1064039) G73A polymorphism and Alzheimer's disease (AD) is controversial. The objective of the study was to investigate the possible association between CST3 G73A polymorphism and AD. METHOD: We performed a meta-analysis pooling data from all relevant studies including 2,410 cases and 2,539 controls. We applied a random-effects or fixed-effects model to combine odds ratio (OR) and 95% confidence intervals (95% CI). RESULTS: Positive associations of the CST3 G73A polymorphism with AD risk were found in allele comparison A versus G (OR = 1.61, 95% CI = 1.19-2.18), dominant model AG + GG versus AA (OR = 0.63, 95% CI = 0.47-0.86), and homozygote comparison AA versus GG (OR = 1.61, 95% CI = 1.19-2.18). In subgroup analysis stratified by ethnicity, significant associations were also demonstrated in Caucasians under allele comparison (OR = 1.17, 95% CI = 1.02-1.33), dominant genetic (OR = 0.59, 95% CI = 0.40-0.86) model, and homozygote comparison (OR = 1.73, 95% CI = 1.18-2.54), but not in Asians. In subgroup analysis according to the age of onset, 73A allele (A versus G) was significantly associated with susceptibility to AD in Caucasians (OR = 1.26, 95% CI = 1.06-1.50), However, no association was found in Asians. CONCLUSION: The CST3 G73A polymorphism is associated with AD in Caucasian populations, but not in Asians.