Liver Cancer Stage Research Articles

LncRNA RHPN1-AS1 Promotes Cell Proliferation, Migration and Invasion Through Targeting miR-7-5p and Activating PI3K/AKT/mTOR Pathway in Hepatocellular Carcinoma.

Hepatocellular carcinoma (HCC) is a severe disease with high mortality in the world. Emerging evidence has suggested that lncRNAs play an important role in cancer progression, including HCC. This study aimed to comprehensively investigate the effect of lncRNA RHPN1 antisense RNA 1 (RHPN1-AS1) on HCC and its underlying molecular mechanism. In this study, we evaluated the expressions of lncRNA RHPN1-AS1 and miR-7-5p by qRT-RCR in both HCC tissue and HCC cells. Our findings showed that lncRNA RHPN1-AS1 was upregulated in HCC tissue and HCC cells, while miR-7-5p was downregulated. LncRNA RHPN1-AS1 expression in HCC patients was closely related to vascular invasion, tumor-node-metastasis (TNM) stage and barcelona clinic liver cancer (BCLC) stage. Furthermore, we quantified cell clone-formation ability, proliferation, migration and invasion of HCCLM3 and MHCC97 H cells using several assays (colony formation assay, 5-Ethynyl-2′-deoxyuridine (EdU) assay and transwell assay, respectively). Functional experiments confirmed that silencing lncRNA RHPN1-AS1 inhibited cell proliferation, migration and invasion in HCCLM3 and MHCC97 H cells. After that, bioinformatics analysis, dual luciferase reporter gene assay, qRT-PCR and western blot were used to investigate the molecular mechanism of lncRNA RHPN1-AS1 on HCC. Mechanistically, the rescue experiments demonstrated that miR-7-5p inhibitor reversed the inhibition effect of silencing lncRNA RHPN1-AS1 on HCCLM3 cells proliferation, migration and invasion. Moreover, silencing lncRNA RHPN1-AS1 also inhibited the activation of PI3K/AKT/mTOR pathway. Taken together our findings demonstrated that lncRNA RHPN1-AS1 could facilitate cell proliferation, migration and invasion via targeting miR-7-5p and activating PI3K/AKT/mTOR pathway in HCC.

Safe hepatectomy after arterial-based liver directed therapy

Presenter: Aneil Sood | Indiana University Background: Liver directed therapy (LDT) for primary and secondary liver cancer has been increasingly utilized in patients with technically unresectable liver tumors, or in patients with surgically prohibitive underlying liver disease or medical comorbidities. Recent studies have shown that transarterial radioembolization (TARE) may reduce tumor burden and permit hepatectomy for complete tumor clearance. The short- and long-term outcomes of patients undergoing hepatectomy following TARE are reported in this study. Methods: An observational study of patients undergoing partial hepatectomy after TARE with yttrium-90 (Y90) between 2013-2018 was performed. Demographics, comorbidities, TARE specific variables, perioperative outcomes, and long-term outcomes were recorded. Variables specific to TARE included the number of procedures, arterial selectivity of embolization, and total millicurie (mCi) dose. Operative variables included the extent of hepatectomy, estimated blood loss (EBL), duration of operation, major postoperative morbidity (Clavien-Dindo Grade III-V complications), hospital readmission, and 30-day mortality. Long-term outcomes were recorded as pattern of recurrence, disease-free survival, and overall survival. Results: Eight patients (6 females, 2 males; mean age 60.4 +/- 1.8 years) underwent TARE for initially unresectable liver cancer, including hepatocellular carcinoma (n = 5), metastatic cancer (pancreatic neuroendocrine n = 1, pulmonary neuroendocrine n = 1), and cholangiocarcinoma (n = 1). The mean total dose of Y90 was 86.1 +/- 14.4 mCi during a single procedure (n = 4) or over two procedures (n = 4). Selective TARE was performed in five patients (63%) and nonselective in three patients (38%). The mean time from TARE to hepatectomy was 12.4 +/- 4 months. Six of the 8 patients (75%) required major hepatectomy (all open technique). Mean EBL was 1056 +/- 294 mL, and mean operative time was 271 +/- 43 minutes. Postoperatively, major morbidity was seen in three patients (38%) and included organ-space surgical site infection (n = 1), pneumonia requiring reintubation (n = 1), and bile leak (n = 2). Readmission occurred in four patients (50%). There were no 30-day mortalities. After a median follow-up of 37 months, recurrence was observed in four patients (50%); three patients developed hepatic recurrence, and one patient developed peritoneal recurrence. The mean disease-free survival and overall survival were 18.1 +-/ 5.0 months and 33.8 +/- 8.4 months, respectively. Conclusion: Hepatectomy following liver-directed therapy with Y90 radioembolization appears to be a safe treatment paradigm for patients with advanced stage liver cancer and can result in good long-term outcomes.

MRI contrast enhancement of liver pre-neoplasia using iron-tannic nanoparticles.

The most challenging part of liver cancer detection is finding it in the very early stages. It has been argued that liver preneoplasia is found at the very earliest stages of liver cancer. The presence of a lesion is closely related to the development of HCC. We report herein a new class of iron-based T1 MRI contrast agents which are nanoparticles of iron–tannic complexes (so-called Fe–TA NPs) that can be used for detecting liver preneoplasia. Preliminary assessment of their toxicity in healthy rats provides suitable imaging dose ranges with acceptable toxicity. In diethylnitrosamine (DEN) induced rats, it is shown that Fe–TA NPs are capable of enhancing MRI signals in rat livers having pre-neoplastic lesions within 60 minutes post-injection. The enhancement efficacy is strongly dependent on the characteristics of pre-neoplastic foci (GST-P+ foci). The highest enhancement was in good correlation with the size of GST-P+ foci and amount of Fe–TA NPs accumulated in the liver, and might be caused by the dysfunction of liver sinusoids along with cellular uptake capability of pre-neoplastic hepatocytes. Our results show that Fe–TA NPs are of great interest to develop as an efficient MRI imaging agent for risk assessment of liver cancer.

Clothing spiny nanoprobes against the mononuclear phagocyte system clearance in vivo: Photoacoustic diagnosis and photothermal treatment of early stage liver cancer with erythrocyte membrane-camouflaged gold nanostars

Nanoprobes have been increasingly applied in photoacoustic imaging to improve the diagnosis and treatment of diseases, especially the liver cancer, the fourth leading cause of cancer-related deaths worldwide. The mononuclear phagocytic system (MPS), however, exhibits a crucial impediment to the probe usage, preventing the accumulation of probes within the liver. This, in turn, paralyzes the effort of diagnosis enhancement. To overcome the MPS clearance, a biomimetic probe consisting of erythrocyte membrane-camouflaged gold nanostars was designed and synthesized in this study. The probe possesses broad absorption spectrum, photostability, and photothermal conversion efficiency. More importantly, it yields preeminent immune escape ability, thus being able to escape the MPS clearance and greatly boost the accumulation of probes at the tumor sites with prolonged blood circulation. Experimentally, we demonstrate that the probe can serve as an effective nanoprobe to enhance the in vivo photoacoustic imaging and photothermal treatment of in situ early stage liver tumors in mice.

Prognostic Nomogram for Hepatitis B Virus-related Hepatocellular Carcinoma With Adjuvant Transarterial Chemoembolization After Radical Resection.

The objective of this study was to establish a reliable and effective nomogram for predicting prognosis of hepatitis B virus (HBV)-related hepatocellular carcinoma (HCC) with postoperative adjuvant transarterial chemoembolization (TACE). A derivation cohort of 370 HCC patients treated with postoperative TACE in the Eastern Hepatobiliary Surgery Hospital from January 2009 to December 2012 were retrospectively analyzed. Univariate and multivariate analysis were performed by Cox regression and independent prognostic factors for overall survival were determined to construct the nomogram. Concordance index (C-index), calibration curve and decision curve analysis were performed to evaluate the capability of the nomogram and the established nomogram was compared with TNM stage and Barcelona Clinic Liver Cancer (BCLC) stage to identify the superior model. The results were validated in a validation cohort of 123 HCC patients in the same center. Multivariate analysis indicated that γ-glutamyl transferase, α-fetoprotein, tumor number, tumor size, satellite lesions, microvascular invasion, and HBV-DNA were independent prognostic factors for overall survival in the derivation cohort, and all these factors were selected into the nomogram. The C-index was 0.755 for survival prediction of the nomogram, which was significantly higher than the TNM stage (0.636, P<0.001) and BCLC stage (0.594, P<0.001). A fair uniformity and a superior net benefit with wide range threshold probabilities were showed in the calibration curves and decision curve analysis. In the validation cohort, the C-index of the nomogram (0.785) also had a higher predictive accuracy than TNM stage (0.744, P=0.019) and BCLC stage (0.616, P<0.001). The nomogram with accurate and reasonable performance was proposed for predicting survival of HBV-related HCC with postoperative adjuvant TACE.

Impact of overweightness and critical weight loss on overall survival in patients with hepatocellular carcinoma initially treated with chemoembolization.

BackgroundThe effects of overweightness and weight loss on the development and prognosis of hepatocellular carcinoma (HCC) remain unclear. In this study, we aimed to evaluate the impact of overweightness and weight loss on the survival of patients with intermediate/advanced HCC receiving chemoembolization as initial treatment.MethodsWe examined 1,170 patients who underwent chemoembolization as initial treatment for Barcelona-Clínic Liver Cancer stages B and C HCC at Sun Yat-sen University Cancer Center (Guangzhou, China) between December 2009 and May 2015. A baseline body mass index (BMI) of ≥23 kg/m2 was defined as overweight, and body-weight loss of ≥5.0% from baseline was defined as critical weight loss (CWL). Cox regression analysis was used to determine the association between overweightness or CWL and overall survival (OS).ResultsThe median survival time was 16.8 (95% confidence interval, 13.9–19.7) months and 11.1 (95% confidence interval, 10.0–12.2) months in the overweight and non-overweight groups (log-rank test, P < 0.001), respectively. Cox multivariate analysis identified overweightness as an independent protective prognostic factor for OS (P < 0.001). Subgroup stratification analysis revealed a significant association between overweightness and survival among patients receiving further treatment (P = 0.005), but not in those not receiving further treatment (P = 0.683). Multivariate analysis showed that both overweightness and CWL were independent prognostic factors for OS among patients receiving further treatment.ConclusionAmong patients with intermediate- or advanced-stage HCC initially treated with chemoembolization, overweightness was associated with longer OS. Furthermore, CWL was an independent adverse prognostic factor for OS in patients receiving additional treatment.

Comparison of the effect on the prognosis of HCC in terms of different surgical approaches for hepatic inflow occlusion

Background: To investigate the effect of surgical approaches for hepatic inflow occlusion performed during hepatectomy on postoperative tumor-free survival (TFS) in patients treated with R0 resection. Methods: In total, 343 hepatocellular carcinoma (HCC) patients who underwent hepatectomy (R0) with different surgical approaches for hepatic inflow occlusion were analyzed retrospectively. Results: In total, 68.80% of the patients underwent hepatic inflow occlusion, including 46.65% with a routine Pringle maneuver and 22.16% with selective hemi-hepatic vascular exclusion (SHVE), during surgery. The TFS did not significantly differ among the Pringle group, the SHVE group and the no hepatic inflow occlusion group. After further stratifying the patients according to the Barcelona clinic liver cancer (BCLC) staging system, the patients with BCLC stage A disease in the SHVE group and no hepatic inflow occlusion group displayed better TFS than those in the Pringle maneuver group (P=0.04; P=0.002), but the patients with BCLC stage B–C disease did not show significant differences among the groups. Furthermore, all 214 patients with BCLC stage A disease were classified into two subgroups according to the microvascular invasion (MVI) status. Interestingly, among the patients with MVI, those in the SHVE group and no hepatic inflow occlusion group had significantly longer TFS than those in the Pringle group (P=0.025; P=0.006); however, the patients without MVI did not show differences among the groups. Additionally, the multivariate analysis revealed that a tumor size ≥5 cm, an absent capsule, a low Edmondson grade, MVI positivity and performance of the Pringle maneuver were independent risk factors of the prognosis in patients with BCLC stage A disease. Conclusions: Hepatic inflow occlusion and the surgical approach used to address this issue may have an impact on HCC prognosis in patients with BCLC stage A disease, especially among those with MVI positivity. No hepatic inflow occlusion or an SHVE approach rather than the Pringle maneuver should be considered first during hepatectomy for patients with BCLC stage A disease.

Rap GTPase Interactor: A Potential Marker for Cancer Prognosis Following Kidney Transplantation.

Post-transplant (post-Tx) kidney cancer has become the second-highest cause of death in kidney recipients. Late diagnosis and treatment are the main reasons for high mortality. Further research into early diagnosis and potential treatment is therefore required. In this current study, through genome-wide RNA-Seq profile analysis of post-Tx malignant blood samples and post-Tx non-malignant control blood samples (CTRL-Tx), we found Rap GTPase Interactor (RADIL) and Aprataxin (APTX) to be the most meaningful markers for cancer diagnosis. Receiver operating characteristic (ROC) curve analysis showed that the area under the curve (AUC) of the RADIL-APTX signature model was 0.92 (P < 0.0001). Similarly, the AUC of RADIL alone was 0.91 (P < 0.0001) and that of APTX was 0.81 (P = 0.001). Additionally, using a semi-supervised method, we found that RADIL alone could better predict malignancies in kidney transplantation recipients than APTX alone. Kaplan-Meier analysis indicated that RADIL was expressed significantly higher in the early stages (I and II) of kidney, liver, stomach, and pancreatic cancer, suggesting the potential use of RADIL in early diagnosis. Multivariable Cox regression analysis found that RADIL together with other factors (including age, stage III, stage IV and CD8+ T cells) play a key role in kidney cancer development. Among those factors, RADIL could promote kidney cancer development (HR > 1, P < 0.05) while CD8+ T cells could inhibit kidney cancer development (HR < 1, P < 0.05). RADIL may be a new immunotherapy target for kidney cancer post kidney transplantation.

Abstract 837: Circulating cell-free DNA methylation assay: Towards early detection of multiple cancer types

Abstract Background: Alterations in DNA methylation play a key role in tumor initiation, and methylation markers are well established for detecting various cancer types. Cancer-specific DNA methylation patterns of cell-free DNA (cfDNA) isolated from blood samples is a non-invasive method to obtain representative epigenetic information from solid tumors. In the present study, we identified and validated cancer-specific methylation markers for diagnosis of liver, breast and colorectal cancers with high sensitivity and specificity. Methods: Three blinded validation studies were performed in order to evaluate individual panels of DNA methylation markers developed for the detection of liver, breast or colorectal cancers. For the liver cancer panel study, samples drawn from 154 subjects were analyzed, including healthy donors (30) and subjects diagnosed with benign liver disease (10), liver cancer stage I-IV (60), or another cancer type (30). For the breast cancer panel study, samples drawn from 151 subjects were analyzed, including healthy donors (39) and subjects diagnosed with benign breast disease (15), breast cancer stage I-IV (65), or another cancer type (32). For the colorectal cancer panel study, samples drawn from 154 subjects were analyzed, including healthy donors (42) and subjects diagnosed with benign colorectal disease (14), colorectal cancer stage I-IV (68), or another cancer type (30). Cell-free DNA was extracted from all samples, bisulfite converted, and DNA methylation was quantified at target sites by using the IvyGene Platform for each of the three cancer-specific marker panel. Results: By quantifying DNA methylation at the target sites, the cancer-specific markers were able to differentiate subjects diagnosed with cancer from both healthy donors and subjects with benign diseases. The liver cancer cfDNA methylation panel showed an overall sensitivity of 95% and specificity of 97.5%. Whereas, the breast cancer cfDNA methylation panel correctly identified the breast cancer samples for an overall calculated sensitivity of 89% and specificity of 96%. The colorectal cancer cfDNA methylation panel also showed high overall sensitivity of 93% and specificity of 100%. Conclusions: These results demonstrate the high diagnostic potential of cfDNA methylation markers isolated from blood for the detection of multiple cancers at various stages with high sensitivity and specificity. In addition, a quantitative analysis of cfDNA methylation signature provides an opportunity for monitoring of disease. The benefits of a sensitive and specific assay to identify and monitor cancer is anticipated to aid disease management and ultimately, improve patient outcomes. Citation Format: Dhruvajyoti Roy, David Taggart, Lianghong Zheng, Dan Liu, Gen Li, Mingzhen Li, Kang Zhang, Richard A. Van Etten. Circulating cell-free DNA methylation assay: Towards early detection of multiple cancer types [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 837.

Abstract CT175: HEPANOVA Phase II study design for advanced hepatocellular carcinoma: Tumor treating fields concomitant with sorafenib

Abstract Background Tumor Treating Fields (TTFields) are a non-invasive, regional antimitotic treatment modality, which has been approved for the treatment of glioblastoma by the FDA. TTFields predominantly act by disrupting the formation of the mitotic spindle during metaphase. TTFields were effective in multiple preclinical models of hepatocellular carcinoma (HCC), leading to a significant increase in cell death. The Phase II HEPANOVA study is the first trial testing TTFields in HCC patients, and is designed to test the safety and efficacy of adding TTFields to sorafenib in advanced HCC. Methods Trial Design: Patients (N=25) with unresectable HCC who are not amenable to any local treatment will be enrolled in this prospective, single-arm study. The study enrolls patients with ECOG score of 0-2 and Barcelona clinic liver cancer (BCLC) stage 0-C. Patients must have a measurable disease per RECIST Criteria. Having implanted electronic devices in the torso is exclusionary. Sorafenib will be administered at standard dose (400 mg daily). TTFields (150 kHz) will be delivered for 18 hours/day until local disease progression per RECIST Criteria. Clinical follow up will be performed q4w, and a CT/MRI scan q12w. Following disease progression in the liver, patients will discontinue TTFields and be followed monthly for survival. Overall response rate will be the primary endpoint and in-field control rate, progression-free survival rate at 12 month (PFS12), OS rate at 1 year and toxicity will all be secondary endpoints. Sample size was calculated using an Exact test for proportions considering the weighted average of ORR of patients who had either complete or partial response per RECIST criteria in historical studies with sorafenib is 4.5%. A sample size of 25 patients was required to achieve a power of approximately 80% at a one-sided alpha level of 0.05 using a single sample Exact test for proportions. Citation Format: Anca-Ligia Grosu, Eleni Gkika, Thomas Brunner, Robert Thimme, Uri Weinberg. HEPANOVA Phase II study design for advanced hepatocellular carcinoma: Tumor treating fields concomitant with sorafenib [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr CT175.

Abstract 2311: Analysis of glypican 3-targeted chimeric antigen receptor T cells in hepatocellular carcinoma cell and mouse models

Abstract Background: Glypican-3 (GPC3) is a cell surface protein that is highly expressed in hepatocellular carcinoma (HCC). Methods and Results: In this report, we analyzed two chimeric antigen receptors (CARs) targeting GPC3, CAR (HN3) recognizes the N-lobe of GPC3 while CAR (hYP7) targets the C-lobe that is close to the cell surface. In a real-time cell cytolytic assay, we found that CAR (hYP7) T cells showed higher cytolytic activity than CAR(HN3) T cells at an E:T ratio of 2:1 when co-cultured with HepG2 cells for up to 140 hours. The Luminex analysis showed that CAR (hYP7) T cells produced significantly more Th1/Th2 cytokines and chemokines than CAR (HN3) T cells following exposure to Hep3B and HepG2 cells. In the Hep3B peritoneal dissemination xenograft mouse model, a single intraperitoneal infusion of CAR (hYP7) cells exhibited sustainable antitumor efficacy, and all of the mice survived without recurrence 10 weeks post treatment. Strikingly, we demonstrated that single injection of CAR (hYP7) T cells derived from either healthy donor or HCC patient significantly regressed tumor growth in the orthotopic HCC mouse model. Using a highly sensitive droplet digital PCR (ddPCR), we detected high frequencies of CAR T cells in the spleen and tumor microenvironment of mice in the CAR (hYP7) group. By contrast, CD19 CAR T cells showed no sign of gene integration in either tissue. More importantly, we found that CD8+ CAR (hYP7) T cells recovered from mouse spleens exhibited much higher lytic activity against HCC cells than CD4+ CAR (hYP7) T cells. Finally, we found that the GPC3-targeted CAR T cells downregulated the Wnt signaling in HCC cells. Conclusion: The hYP7 antibody-derived GPC3-specific CAR is a promising therapeutic that could be tested for the treatment of advanced stage liver cancer. Citation Format: Dan Li, Nan Li, Yifan Zhang, Haiying Fu, Zhijian Duan, Alissa Hummer, Hongjia Yang, Madeline B. Torres, Xiaoling Luo, Ling Su, Hu Zhu, Josh Kramer, Jinqiu Chen, Xiaolin Wu, Matthew Hall, Qun Wang, Stephen Hewitt, Tim Greten, Mitchell Ho. Analysis of glypican 3-targeted chimeric antigen receptor T cells in hepatocellular carcinoma cell and mouse models [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 2311.

Evaluation of Fucosylated Haptoglobin as a Diagnostic Biomarker for Hepatocellular Carcinoma in Egypt

Background: Most Hepatocellular Carcinomas (HCCs) are diagnosed at an advanced stage. Therefore, there is citation-type an urgent need for better methods of detection and surveillance of patients at risk of HCC. Alpha-fetoprotein (AFP) has a suboptimal diagnostic performance for HCC surveillance, so novel and reliable diagnostic biomarkers are required. Objective: The aim of this work is to evaluate fucosylated haptoglobin as a diagnostic biomarker for hepatocellular carcinoma in Egyptian patients. Materials and Methods: This case-control study was carried out on 60 patients classified into 3 groups (20 patients on each); group I (HCC group), group II (Cirrhotic group) and group III (Control group). Diagnosis of liver cirrhosis was done by clinical, biochemical and ultrasound (US), whereas the diagnosis of HCC was done by percutaneous biopsy or radiological (by US and triphasic Computerized Tomography (CT) based on the guidelines of the American-Association for the Study of Liver Diseases. HCC stage was clinically defined according to the Barcelona Clinic Liver Cancer (BCLC) staging system. AFP &amp; fucosylated haptoglobin levels were estimated in all groups. Results: There was a statistically significant positive correlation between serum fucosylated haptoglobin and tumor size in the HCC group. Serum fucosylated haptoglobin (at 116 U/ ml) showed sensitivity 80%, specificity 65%, positive predictive value 53% and negative predictive value 87% with AUC 0.786. Combination of serum fucosylated haptoglobin and serum AFP at (200 ng/ ml) increased sensitivity that reached 95%. Conclusion: Serum fucosylated haptoglobin may serve as a novel diagnostic biomarker for the detection of HCC with higher sensitivity than AFP.

MicroRNA-548b suppresses aggressive phenotypes of hepatocellular carcinoma by directly targeting high-mobility group box 1 mRNA.

Background and purpose: An increasing number of studies have revealed that microRNAs (miRNAs) are the main drivers of hepatocarcinogenesis including progression to later stages of liver cancer. Recently, miR-548b was identified as a cancer-related miRNA in glioma and tongue squamous cell carcinoma. Nonetheless, the expression pattern and specific roles of miR-548b in hepatocellular carcinoma (HCC) have not yet been clarified.Methods: Expression levels of miR-548b in HCC tissues and cell lines were measured by reverse-transcription quantitative PCR. In vitro and in vivo functional assays were performed to determine the effects of miR-548b on the malignant phenotypes of HCC cells. In addition, the molecular mechanisms by which miR-548b regulates the initiation and progression of HCC were investigated in detail.Results: miR-548b expression was weak in HCC tissues and cell lines. The low miR-548b expression significantly correlated with tumor size, TNM stage, and venous infiltration of HCC. In addition, exogenous miR-548b expression suppressed HCC cell proliferation, colony formation, and metastasis and induced apoptosis in vitro. Silencing of miR-548b exerted an opposite effect on these characteristics of HCC cells. Furthermore, miR-548b overexpression hindered tumor growth in vivo. Mechanistic analysis identified high-mobility group box 1 (HMGB1) as a direct target gene of miR-548b in HCC cells. Moreover, an HMGB1 knockdown reproduced the effects of miR-548b upregulation on HCC cells. Recovered HMGB1 expression reversed the effects of miR-548b on HCC cells. Notably, miR-548b overexpression deactivated the PI3K–AKT pathway in HCC cells in vitro and in vivo.Conclusion: Our findings provide the first evidence that miR-548b restrains HCC progression, at least partially, by downregulating HMGB1 and deactivating the PI3K–AKT pathway. Thus, miR-548b might be a novel target for the development of new therapies for HCC.

Underlying liver disease and advanced stage liver cancer are associated with elevated neutrophil-lymphocyte ratio.

Background/AimsInflammation-based scores, such as the neutrophil-to-lymphocyte ratio (NLR), have been associated with prognosis in hepatocellular carcinoma (HCC); but variable cut-off values and potential lack of specificity have limited the utility of NLR. This study evaluates NLR in a large cohort of HCC patients.MethodsWe retrospectively reviewed 789 HCC cases (1993–2017) for demographics, tumor characteristics, treatment, and survival. NLR was stratified into NLR ≥1.5 and NLR ≥3 and analyzed for correlation with American Joint Committee on Cancer (AJCC) and Barcelona Clinic Liver Cancer (BCLC) stages. In 235 patients who underwent liver resection, survival and recurrence were evaluated by NLR.ResultsIn 789 HCC cases, mean NLR was increased with advanced AJCC and BCLC stages. Hepatitis C patients were less likely to have NLR ≥1.5 and ≥3. Non-alcoholic steatohepatitis patients were more likely to have NLR ≥3. Patients with tumor size >5 cm, rupture, or macrovascular invasion were more likely to have NLR ≥3. In patients treated with resection, NLR ≥3 predicted early recurrence (odds ratio [OR] 4.14, P<0.01) and overall recurrence (OR 4.05, P<0.01). Mean NLR was 4.30 in those with recurrence and 2.75 in those without recurrence. Patients with NLR ≥3 showed significantly worse survival compared to those with NLR <3 (P<0.01 by log-rank test).ConclusionsElevated NLR is associated with advanced cancer stage and aggressive tumor characteristics, such as large size, rupture, and invasion. NLR ≥3 was associated with early and overall recurrence after resection but varied with etiology. NLR may be a useful biomarker in predicting recurrence for HCC patients undergoing curative resection, but further studies are required to elucidate the effect of disease etiology.

03:27 PM Abstract No. 366 Quantification of the heat-sink effect during microwave ablation with 4D-flow MRI in an in-vivo porcine liver model

Image-guided thermal ablation has become the standard of care for treating early stage liver cancer. However, a significant limitation to a successful ablation procedure is the heat-sink effect, which may lead to suboptimal thermal dose delivery. Currently, there is no reliable method for quantifying the heat-sink effect due to limited information on blood vessel flow and its associated anatomy relative to the ablation zone. The purpose of this study was to use 4D-flow MRI to map out liver flow patterns and correlate flow rates with ablation zone volumes. Female domestic swine (n=3) were anesthetized and prepared as per IACUC guidelines. A subject-specific map of the hepatic vasculature was generated via 4D-flow MRI. Microwave ablation zones were created in individual liver lobes near major vessels by applying 65 W for 5 minutes through a high-powered microwave antenna. For each ablation zone, the pre-ablation and post-ablation flow rates (mL/sec) of the encompassed vessels, as well as their corresponding ablation sizes (mL) were recorded and analyzed via logistic regression modeling and Student t-test. A total of n=9 ablation zones were created that encompassed individual portal and hepatic veins. There were 3/6 (50%) thrombosed portal veins and 0/8 (0%) thrombosed hepatic veins (p=0.21). Hepatic vein flow rate decreased significantly after ablation (2.3±0.7 mL/sec vs 1.5 ±0.4 mL/sec, p=0.01). Portal vein flow rate similarly decreased (2.1±0.9 mL/sec vs 1.8±1.1 mL/sec, p=0.72) after ablation, although not significantly so. Ablation zones that encompassed vessels with flow rates > 2.0 mL/sec were significantly smaller in volume than ablation zones encompassing vessels with flow rates < 2.0 mL/ sec (4.2±3.0 mL vs 9.3±2.5 mL, p=0.03). Microwave ablation zones were found to be significantly smaller in volume when encompassing vessels with flow rates greater than 2.0 mL/sec. Both portal veins and hepatic veins show decreased flow in ablation zones, likely relating to their response to thermal damage. Mapping out blood flow vasculature with 4D-flow MRI prior to ablation zones can assist clinicians in deciding how aggressively to treat liver tumors.

Effect of Nucleos(t)ide Analogs on Patients with Intermediate and Advanced Hepatitis B Virus-Related Hepatocellular Carcinoma.

The role of nucleos(t)ide analogs (NAs) therapy in intermediate and advanced hepatitis B virus (HBV)-related hepatocellular carcinoma (HCC) remains unclear. The aim was to evaluate the effect of NAs therapy on survival of intermediate- and advanced-stage HBV-related HCC patients initially treated with chemoembolization. A total of 1016 Barcelona Clinic Liver Cancer (BCLC) stage B/C HBV-related HCC patients initially treated with chemoembolization were included. Propensity score matching (PSM) was performed to decrease heterogeneity between the antiviral and non-antiviral groups. Kaplan-Meier and Cox regression analysis were performed to evaluate the effects of NAs therapy on overall survival (OS). Antiviral group (n = 394) significantly prolonged OS compared with non-antiviral group (n = 622) (p = 0.003). NAs therapy (p < 0.001) along with tumor size (p = 0.002), tumor number (p = 0.001), gross vascular invasion (p < 0.001), metastasis (p < 0.001), α-fetoprotein (p < 0.001), Child-Pugh score (p = 0.008), aspartate aminotransferase (p < 0.001), and HBV DNA (p = 0.018) were identified as independent prognostic factors for OS. After PSM processing, deducting the influence of subsequent treatments for HCC, NAs therapy was still identified as an independent protective factor (p = 0.009) for OS in patients who survived ≥ 7months, regardless of BCLC stage B or C HCC. NAs therapy prolongs OS in intermediate- and advanced-stage HBV-related HCC patients initially treated with chemoembolization. After PSM processing, patients who survived ≥ 7months still benefited from NAs therapy.

Liver Cancer Detection

Machine learning techniques play an important role in building predictive models by learning from Electronic Health Records (EHR). Predictive models building from Electronic Health Records still remains as a challenge as the clinical healthcare data is complex in nature and analysing such data is a difficult task. This paper proposes prediction models built using random forest ensemble by using three different classifiers viz. J48, C4.5 and Naïve Bayes classifiers. The proposed random forest ensemble was used for classifying four stages of liver cancer. Using a feature selection method the reliable features are identified and this subset serves as input for the ensemble of classifiers. Further a majority voting mechanism is used to predict the class labels of the liver cancer data. Experiments were conducted by varying the number of decision trees generated using the J48, C4.5 and Naïve Bayes classifiers and compared with the classification made using decision stump and Adaboost algorithms.

Neutrophil to Lymphocyte Ratio as Predictive Marker of Recurrence of HCC after Radiofrequency Ablation

Background: Neutrophil to lymphocyte ratio (NLR), platelet to lymphocyte ratio (PLR), and prognostic nutritional index (PNI), are immunonutritional indices, have been shown as an independent factor to predict postoperative recurrence and/or overall survival in patients with hepatocellular carcinoma (HCC). Aim: to validate neutrophils-to-lymphocytes ratio (NLR) as a predictor of post radiofrequency ablation recurrence of hepatocellular carcinoma (HCC). Patients and methods: This cross sectional prospective study was conducted on 50 Egyptian patients had radiofrequency ablation for HCC within Barcelona clinical liver cancer (BCLC) stage A. Pre-treatment laboratory tests and imaging were used to measure NLR, PLR, PNI, Child-Pugh (CTP score, tumor number and tumor size. HCC recurrences were followed after three month. Results: According to NLR cut-off value, the patients with NLR >1.73 had larger tumour size, and higher rates of tumor multiplicity .After 3 month follow up 30/50 (60%) patients had tumour recurrence. Regarding recurrence after RFA showed that CTP class B (P 11.74), NLR-PNI 1 group (NLR > 1.73 or PNI ≤ 11.74) and NLR-PNI 2 group (NLR > 1.73 and PNI ≤ 11.74). The patients with NLR-PNI 2 group had increase in number of portahepatis lymph node, number and size of tumor, decrease serum albumin and change in PNLR. Multivariate analyses suggested increased NLR (hazard ratio [HR] =2.09; 95% confidence interval [CI] =1.88–2.55; PConclusion: High blood NLR after RFA is a predictor for worse survival and also can predict recurrence of HCC. Higher NLR-PNI score predict a worse prognosis in patients who underwent RFA.

Safety and efficacy of cytotoxic chemotherapy in hepatocellular carcinoma after first-line treatment with sorafenib

BackgroundBefore the targeted therapies era, cytotoxic chemotherapy (CCT) was an option for advanced hepatocellular carcinoma (HCC), even with the lack of supporting evidence. Since the last decade, sorafenib has been established as the first-line therapy. Although new agents are being incorporated, CCT is still considered in regions where new drugs are not available or for patients who progressed through the approved therapies and remain in good clinical condition. We aimed to describe our experience regarding the use of CCT as second-line treatment after sorafenib.MethodsA database of 273 patients was evaluated. Patients that received CCT after sorafenib progression were selected for the analysis. Descriptive statistics was used for categorical and continue variables. Median survival was estimated with Kaplan-Meier curves. Variables were found to be significant if the two-sided p value was ≤ 0.05 on multivariate testing using the Cox regression model.ResultsForty-five patients received CCT; 33 (73.3%) had Child-Pugh classification A, and 34 (75.6%) had stage C according to the Barcelona Clinic Liver Cancer (BCLC) staging system. The most used regimen was doxorubicin in 25 patients (55.6%). Median overall survival (OS) was 8.05 months (95% confidence interval [CI] 2.73 – 9.88 months). The 6-month and 1-year survival probability was 52.4% and 27.36%, respectively. Eastern Cooperative Oncology Group performance status (ECOG PS) 0–1 and disease control with sorafenib was independently associated with better OS in patients treated with CCT. Any-grade toxicities were observed in 82.2% and grade 3–4 in 44.4% of the patients.ConclusionIn accordance with previous studies, CCT had a notable rate of adverse events. The poor prognosis of this cohort suggests that CCT may not alter the natural history of HCC after sorafenib progression.

MicroRNA‑363‑3p inhibits hepatocarcinogenesis by targeting HMGA2 and is associated with liver cancer stage.

The importance of microRNAs (miRNAs) in cancer development has been widely recognized in recent decades. In the present study, the function and mechanism of miRNA-363-3p (miR-363-3p), formerly characterized as a tumor suppressor, in the hepatocarcinogenesis of liver cancer cells was investigated. Reverse transcription-quantitative polymerase chain reaction (RT-qPCR) was applied to detect the expression of miR-363-3p in liver cancer tissues. Cell proliferation, survival and migration capacities were determined by MTT, colony formation and wound-healing assays, respectively. The targeting of high mobility group AT-hook 2 (HMGA2) mRNA by miR-363-3p was confirmed by bioinformatics analysis, and RT-qPCR, luciferase reporter and western blot assays. The correlation between the expression levels of HMGA2 and miR-363-3p was analyzed. The RT-qPCR results revealed that the levels of miR-363-3p were downregulated in liver cancer tissues. Cellular assays validated that miR-363-3p exerted tumor suppressing functions, including the inhibition of cell proliferation, survival and migration abilities in two liver cancer cell lines. Bioinformatics prediction and subsequent experiments demonstrated that HMGA2 was a direct target of miR-363-3p. Restoration of the expression of HMGA2 in miR-363-3p mimic-transfected cells reversed the tumor suppressing effects caused by miR-363-3p. Finally, there was a significant negative correlation between the expression levels of HMGA2 and miR-363-3p in liver cancer tissues. miR-363-3p was identified as an important tumor suppressor in liver cancer via targeting HMGA2, which may have potential benefits in liver cancer therapy.