Local Recurrence Of Hepatocellular Carcinoma Research Articles

A first-in-human study to evaluate a personalized neoantigen-based mRNA-loaded dendritic cell vaccine, in combination with ablation, in patients with hepatocellular carcinoma.

e14513 Background: Local recurrence of Hepatocellular Carcinoma (HCC) following potentially curative resection or ablation is an inevitable risk. T cells recognizing neoantigens are present in most cancers including HCC and offer a specific and highly immunogenic target for the personalized vaccination as an adjuvant therapy. The purpose of this study is to evaluate the safety and T cell immune response of a personalized neoantigen-based mRNA loaded dendritic cell (DC) vaccine (LK101) combined with regular ablation in patients with HCC (NCT03674073). Methods: Each patient’s fresh tumor biopsy and matched peripheral blood sample were subjected to whole-exome sequencing and RNA-sequencing. Following determination of somatic mutations and gene expression, up to 30 in silico–predicted potential neoantigens were selected with bioinformatic tool. DCs were generated from the monocytes enriched by leukapheresis, followed by transfection with neoantigen-encoding mRNA through electroporation. In total, 7 doses of LK101 were administered at 1.5×107 cells/injection intradermally with 4-dose priming weekly and 3 boosters in 4-week intervals. The primary endpoint of the clinical study was safety, and key secondary endpoints were immunogenicity, overall survival and recurrence. 24 HCC patients at HKLC stage IIa were enrolled 1:1 to ablation control arm and ablation combined with DC (LK101) arm. Results: The study was completed, patients were followed up for a median of 48.4 months and 38.8 months in vaccination arm and control arm. All of adverse events were Grade 1-2. Most frequent treatment-related adverse events are injection site reactions, inclusive of local pain (63.6%), itch (27.3%) and others (27.3%), such as redness and edema. 3 patients in ablation control arm died, whereas all vaccinated patients survived. The 1-year and 2-year recurrence rates in vaccination arm compared with control arm are 18.2%, respectively. LK101 induced neoantigen-specific T cell responses were investigated by ELISpot analysis. Overall, all patients elicited at least one measurable response. Immune responses to 60-90% of neoantigen peptides were seen post vaccination across patients. Both preexisting immune response and De Novo immune response that targeted neoantigens were observed in vaccinated patients. Moreover, both CD4 and CD8 positive effector memory T cells showed the trend of increase, and PD-1 positive T cells reduced in peripheral blood of patients after vaccination. Conclusions: Combination of LK101 with ablation therapy has a manageable safety profile, showing the evidence of immune activation and potential of prolonged survival. In combination with regular ablation, neoantigen based mRNA loaded DC vaccine is a promising immunotherapy for the treatment of HCC. Clinical trial information: NCT03674073 .

An Investigative Analysis of Therapeutic Strategies in Hepatocellular Carcinoma: A Raetrospective Examination of 23 Biopsy-Confirmed Cases Emphasizing the Significance of Histopathological Insights

Background: The Liver Imaging Reporting and Data System (LI-RADS) combines standardized terminology with a classification system for imaging findings in patients with HCC, therefore rendering diagnostic biopsy unnecessary in many cases. This retrospective study included 23 patients with a biopsy diagnosis of HCC, performed either before or after local interventional procedures, in order to evaluate the histopathologic changes induced by previous procedures and their potential influence on the response to immune therapy. Material and Methods: The study encompassed a cohort of patients diagnosed with Hepatocellular Carcinoma (HCC). Diagnosis was established via contrast-enhanced computer tomography or magnetic resonance imaging that identified LI-RADS-5 nodules in conjunction with historical liver disease and elevated alpha-fetoprotein (AFP) levels or via histological examination confirming positivity for glypican3, heat shock protein 70, and glutamine synthetase. The study detailed the liver disease etiology, LI-RADS scores, characteristics and dimensions of HCC nodules, serum AFP concentrations, Edmondson–Steiner grading, and the expression of programmed cell death ligand 1 (PD-L1) in the tumor cells. Results: Among the study’s cohort of Hepatocellular Carcinoma (HCC) patients, a portion had not received any prior treatments, while the remainder experienced local HCC recurrence following trans-arterial chemoembolization or radiofrequency ablation. Observations indicated elevated alpha-fetoprotein (AFP) levels in those who had not undergone any previous interventions, showing statistical significance. The Edmondson–Steiner classification predominantly identified grade III differentiation across patients, irrespective of their treatment history. Furthermore, an increase in intra-tumoral programmed cell death ligand 1 (PD-L1) expression was noted in patients who had not been subjected to previous therapies. Conclusion: Liver biopsy offers valuable insights for patients with Hepatocellular Carcinoma (HCC), assisting in the tailoring of immune therapy strategies, particularly in cases of recurrence following prior local interventions.

Liver Transplantation Outcomes of HBV-, HCV-, and Alcohol-induced Hepatocellular Carcinoma in the United States: Analysis of National Inpatient Samples.

Liver transplantation is a current treatment option for hepatocellular carcinoma (HCC). The United States National Inpatient Sample database was utilized to identify risk factors that influence the outcome of liver transplantation, including locoregional recurrence, distant metastasis, and in-hospital mortality, in HCC patients with concurrent hepatitis B infection, hepatitis C infection, or alcoholic cirrhosis. This retrospective cohort study included HCC patients (n=2391) from the National Inpatient Sample database who underwent liver transplantation and were diagnosed with hepatitis B or C virus infection, co-infection with hepatitis B and C, or alcoholic cirrhosis of the liver between 2005 and 2014. Associations between HCC etiology and post-transplant outcomes were examined with multivariate analysis models. Liver cirrhosis was due to alcohol in 10.5% of patients, hepatitis B in 6.6%, hepatitis C in 10.8%, and combined hepatitis B and C infection in 24.3%. Distant metastasis was found in 16.7% of patients infected with hepatitis B and 9% of hepatitis C patients. Local recurrence of HCC was significantly more likely to occur in patients with hepatitis B than in those with alcohol-induced disease. After liver transplantation, patients with hepatitis B infection have a higher risk of local recurrence and distant metastasis. Postoperative care and patient tracking are essential for liver transplant patients with hepatitis B infection.

Roles of hypoxia-inducible factor in hepatocellular carcinoma under local ablation therapies.

Hepatocellular carcinoma (HCC) is one of the most common digestive malignancies. HCC It ranges as the fifth most common cause of cancer mortality worldwide. While The prognosis of metastatic or advanced HCC is still quite poor. Recently, locoregional treatment, especially local ablation therapies, plays an important role in the treatment of HCC. Radiofrequency ablation (RFA) and high-intensity focused ultrasound (HIFU) ablation are the most common-used methods effective and feasible for treating HCC. However, the molecular mechanisms underlying the actions of ablation in the treatments for HCC and the HCC recurrence after ablation still are poorly understood. Hypoxia-inducible factor (HIF), the key gene switch for adaptive responses to hypoxia, has been found to play an essential role in the rapid aggressive recurrence of HCC after ablation treatment. In this review, we summarized the current evidence of the roles of HIF in the treatment of HCC with ablation. Fifteen relevant studies were included and further analyzed. Among them, three clinical studies suggested that HIF-1α might serve as a crucial role in the RAF treatment of HCC or the local recurrence of HCC after RFA. The remainder included experimental studies demonstrated that HIF-1, 2α might target the different molecules (e.g., BNIP3, CA-IX, and arginase-1) and signaling cascades (e.g., VEGFA/EphA2 pathway), constituting a complex network that promoted HCC invasion and metastasis after ablation. Currently, the inhibitors of HIF have been developed, providing important proof of targeting HIF for the prevention of HCC recurrence after IRFA and HIFU ablation. Further confirmation by prospective clinical and in-depth experimental studies is still warranted to illustrate the effects of HIF in HCC recurrence followed ablation treatment in the future.

Radiofrequency ablation of hepatocellular carcinoma: CT texture analysis of the ablated area to predict local recurrence

PurposeTo investigate the role of CT-texture analysis of liver ablation area to predict local recurrence after HCC ablation. MethodsPatients treated with liver ablation were retrospectively enrolled. CT-texture analysis was performed on the core and borders of ablation area 1–2 months after procedure. Tumors were grouped according to the onset of local recurrence at follow-up (persistence, recurrence-free, short- or long-term recurrence). Differences in texture parameters and which parameters were predictive of recurrence risk were assessed using a Cox regression model. Results151 HCCs were treated in 98 patients (72 ± 9 years, 83 men). 68 HCCs reported no disease recurrence, 32 persistent disease, 19 short-term and 32 long-term recurrence. Median follow-up was 280 [IQR: 156–569] days. Venous phase (Ven)Skewness (HR 6.07, 1.29–28.6, p =.02) and VenKurtosis (HR 2.27, 1.23–4.21, p =.01) of the ablation core were predictive of short-term recurrence. VenHUmean (HR 0.30, 0.11–0.81, p =.02) and VenGLRLM_HGRE (HR 1.06, 1.01–1.11, p =.02) of the core were independent predictors of tumor recurrence (C-index 0.64, CI 0.52–0.76, p =.03). Arterial phase (Art)Entropy of ablation border predicted the recurrence risk (HR 3.15, 1.05–9.42, p =.04) and values higher than 3.71 reported an increased recurrence incidence (p =.05). ArtHUstd (HR 1.14, 1.04–1.24, p =.01), LateHUmean (HR 8.69, 1.11–68.23, p =.04), LateGLRLM_HGRE (HR 0.9, 0.82–0.99, p =.03), LateGLZLM_HGZE (HR 1.01, 1.00–1.02, p <.01) and LateGLZLM_SZHGE (HR 0.99, 0.99–1.00, p =.02) of ablation border were independent predictors of local recurrence risk (C-index 0.73, CI 0.61–0.86, p <.01). ConclusionsCT texture analysis of ablation area performed at 1–2 months follow-up could estimate the risk of local recurrence of hepatocellular carcinoma treated by radiofrequency ablation.

Suppression of Local Tumor Progression in Perivascular Hepatocellular Carcinoma by Combination Therapy with Radiofrequency Ablation and Percutaneous Ethanol Injection: A Propensity Score Matching Analysis

Background and Aims: Radiofrequency ablation (RFA) has replaced percutaneous ethanol injection (PEI) as the treatment of choice for hepatocellular carcinoma (HCC); however, control of local tumor progression (LTP) remains a challenge in perivascular HCC. The aim of this study was to determine whether PEI added to RFA can reduce the LTP rate in perivascular HCC patients. Methods: We retrospectively analyzed 167 patients, with 197 newly diagnosed HCC nodules with peritumoral vessels, who underwent either RFA plus PEI or RFA monotherapy as the first-line treatment between June 2001 and April 2015. Ethanol was injected inside the tumor close to the peritumoral vessels in the combination therapy group. Patients were matched 1:1 according to their propensity scores to reduce selection bias; cumulative LTP was then analyzed using log-rank tests and Cox proportional hazard regression analyses. Results: The two matched groups comprised 62 tumors each. The overall median follow-up period was 34 months (range, 1–140 months). In the RFA plus PEI group, the cumulative LTP rates were 5.7%, 15.5%, and 20.4% at 1, 3, and 5 years, respectively; in the RFA monotherapy group, the rates were 13.2%, 32.0%, and 40.2%, respectively. The rates were significantly lower in the RFA plus PEI group (p = 0.032). Cox proportional hazard regression analysis showed that PEI combination treatment was significantly associated with a reduced risk of local HCC recurrence (hazard ratio, 0.44; 95% confidence interval, 0.19–0.93; p = 0.031). Discussion/Conclusion: The risk of LTP after RFA for perivascular HCC can be significantly reduced by injecting ethanol close to the peritumoral vessels.

Surgical outcomes of pulmonary metastasectomy in hepatocellular carcinoma patients according to approach method: thoracoscopic versus open approach

BackgroundThe role of surgical intervention as a treatment for pulmonary metastasis (PM) from hepatocellular carcinoma (HCC) has not been established. In this study, we investigated the clinical outcomes of pulmonary metastasectomy. Using propensity score matching (PSM) analysis, we compared the results according to the surgical approach: video-assisted thoracic surgery (VATS) versus the open method.MethodsA total of 134 patients (115 men) underwent pulmonary metastasectomy for isolated PM of HCC between January 1998 and December 2010 at Seoul Asan Medical Center. Of these, 84 underwent VATS (VATS group) and 50 underwent thoracotomy or sternotomy (open group). PSM analysis between the groups was used to match them based on the baseline characteristics of the patients.ResultsDuring the median follow-up period of 33.4 months (range, 1.8–112.0), 113 patients (84.3%) experienced recurrence, and 100 patients (74.6%) died of disease progression. There were no overall survival rate, disease-free survival rate, and pulmonary-specific disease-free survival rate differences between the VATS and the open groups (p = 0.521, 0.702, and 0.668, respectively). Multivariate analysis revealed local recurrence of HCC, history of liver cirrhosis, and preoperative alpha-fetoprotein level as independent prognostic factors for overall survival (hazard ratio, 1.729/2.495/2.632, 95% confidence interval 1.142–2.619/1.571–3.963/1.554–4.456; p = 0.010/< 0.001/< 0.001, respectively).ConclusionsMetastasectomy can be considered a potential alternative for selected patients. VATS metastasectomy had outcomes comparable to those of open metastasectomy.

All-trans retinoic acid (ATRA) inhibits insufficient radiofrequency ablation (IRFA)-induced enrichment of tumor-initiating cells in hepatocellular carcinoma.

ObjectiveLocal recurrence of hepatocellular carcinoma (HCC) after radiofrequency ablation (RFA) treatment remains a serious problem. Tumor-initiating cells (TICs) are thought to be responsible for tumor relapse. Here, we investigated the effect of the TIC differentiation inducer, all-trans retinoic acid (ATRA), on RFA and explored the potential molecular mechanisms.MethodsThe proportions of CD133+ and epithelial cell adhesion molecule (EpCAM)+ TICs in recurrent HCC after RFA and primary HCC were first determined in clinic. Then, the effect of heat intervention or insufficient RFA (IRFA) on the malignant potential of HCC cells, including cell migration, sphere formation ability, tumor growth, the proportion of CD133+ and EpCAM+ TICs and expression of stem cell-related genes, was evaluated in vitro andin vivo. Finally, the effect of ATRA on the tumor growth and the proportion of TICs was evaluated. ResultsIn clinical data, a higher proportion of CD133+ and EpCAM+ TICs was found in recurrent tumors than in primary tumors. In vitro heat intervention promoted the cell migration and sphere formation ability. Additionally, it increased the proportion of CD133+ and EpCAM+ TICs and the expression of stem cell-related genes. In addition, after IRFA the residual tumors in xenografts grew faster and had more TICs than untreated tumors. ATRA remarkably inhibited residual tumor growth after IRFA by elimination of TICs though the PI3K/AKT pathway. Combination treatment with ATRA resulted in longer survival outcomes in mouse xenografts than RFA alone. ConclusionsATRA, as a TIC differentiation inducer, could help to improve the effect of RFA treatment, which was partially attributed to its effect against TICs. The data indicated its potential as an alternative drug in the development of better therapeutic strategies for use in combination with RFA.

Monitoring of Heat Shock Response and Phenotypic Changes in Hepatocellular Carcinoma After Heat Treatment

Local recurrence of hepatocellular carcinoma (HCC) after thermal coagulation therapy may be associated with an aggressive phenotypic change. This study focused on the thermal effects on HCC cells and evaluated the heat shock response and phenotypic changes after heat treatment. HepG2 and HuH7 cells were used. After heat treatment at 37-50°C for 5-30 min, we assessed their survival rate, induction of heat shock protein (HSP)70 promoter, proliferation rate, induction of the epithelial-mesenchymal transition (EMT) and cancer stem cell (CSC)-related markers. Induction of HSP70 promoter per surviving cell was maximized after 10 min of heat treatment at 48°C. Induction of EMT and CSC-related markers was also observed. Sub-lethal heat treatment causes large heat shock response to surviving HCC cells and induce EMT-like and CSC-like phenotypic changes that might contribute to increased aggressiveness.

Indicator for local recurrence of hepatocellular carcinoma after proton beam therapy: analysis of attenuation difference between the irradiated tumor and liver parenchyma on contrast enhancement CT.

We aimed to identify dynamic CT features that can be used for prediction of local recurrence of hepatocellular carcinoma (HCC) after proton beam therapy (PBT). We retrospectively retrieved CT scans of patients with PBT-treated HCC, taken between January 2004 and December 2016. 17 recurrent lesions and 34 non-recurrent lesions were retrieved. The attenuation difference between irradiated tumor and irradiated parenchyma (ADHCC-IP) was compared in the two groups by using the Mann-Whitney U test. Cut-off value of ADHCC-IP was estimated by using the Youden index. The follow-up time after PBT initiation ranged from 374 to 2402 days (median, 1069 days) in recurrent lesions, and 418 to 2923 days (median, 1091.5 days) in non-recurrent lesions (p = 0.892). The time until appearance of local recurrence after PBT initiation ranged from 189 to 2270 days (median, 497 days). ADHCC-IP of recurrent lesions [mean, -21.8 Hounsfield units (HU); from -95 to -31 HU] was significantly greater than that of non-recurrent lesions (mean, -51.7 HU; from -117 to -12 HU) at 1-2 years in portal venous phase (p = 0.039). 5-year local tumor control rates were 0.93 and 0.56 in lesions with ADHCC-IP at 1-2 years in PVP < -55 and ≥ -55 HU, respectively. The attenuation difference between irradiated HCC and irradiated liver parenchyma in portal venous phase at 1-2 years after PBT can predict long-term local recurrence of HCC after treatment. We identified a cut-off value for contrast enhancement of HCC after PBT that could predict future local recurrence.

Risk factors for local recurrence of hepatocellular carcinoma after transcatheter arterial chemoembolization with drug-eluting beads (DEB-TACE).

To identify the risk factors for local recurrence in hepatocellular carcinoma (HCC) patients treated with transcatheter arterial chemoembolization (TACE) with drug-eluting beads (DEB-TACE). In this retrospective study, 35 patients (27 males, 8 females; median age 73years) with 116 tumors (median size 14mm) treated with DEB-TACE from May 2014 to September 2018 were evaluated. Age, sex, etiology, Child-Pugh class, alpha-fetoprotein, des-gamma-carboxyprothrombin, previous conventional TACE, tumor location, tumor size, tumor number, contact with the liver surface, level of embolization, corona enhancement on CT during hepatic arteriography, vascular lakes, additional embolization with gelatin sponge particles, and supplying vessels on digital subtraction angiography (DSA) after embolization were analyzed. Univariate analysis showed that advanced age, female, large tumor, contact with the liver surface, and residual supplying vessels were significant risk factors for local recurrence (p = 0.012, 0.0013, 0.0023, 0.025, and < 0.001, respectively). On multivariate logistic analysis, large tumor, contact with the liver surface, and residual supplying vessels on DSA were significant risk factors for local recurrence (p = 0.0026, 0.038, and < 0.001, respectively). Large tumor size, contact with the liver surface, and residual supplying vessels on DSA were significant risk factors associated with local recurrence after DEB-TACE for HCC.

Safety margin of embolized area can reduce local recurrence of hepatocellular carcinoma after superselective transarterial chemoembolization.

Background/AimsWe aimed to determine the relationship between the safety margin of an embolized area and local tumor recurrence (LTR) of patients with hepatocellular carcinoma (HCC) who underwent superselective transarterial chemoembolization (TACE).MethodsThe medical records of 77 HCC patients with 109 HCC nodules who underwent superselective TACE were retrospectively analyzed for LTR. Univariate and multivariate analyses were performed for 16 potential factors using Cox proportional hazard regression. Iodized oil deposition on cone-beam computed tomography (CBCT) imaging was divided into three grades: A=complete tumor staining and complete circumferential safety margin, B=complete tumor staining but incomplete safety margin, C=incomplete tumor staining. The effect of a safety margin on LTR was evaluated by comparison between grade A and B group.ResultsUnivariate and multivariate analyses revealed that grade A iodized oil deposition and portal vein visualization were the only two independent significant factors of LTR (P<0.001 and P=0.029, respectively). The 12- and 24-month LTR rates of tumors for grade A (n=62), grade B (n=30), and grade C (n=17) were 16% vs. 41% vs. 100% and 16% vs. 61% vs. 100%, respectively (P<0.001). The tumors in the grade A group had a 75% risk reduction in LTR (odds ratio, 0.25; 95% confidence interval, 0.10 to 0.64; P=0.004) compared to the grade B group.ConclusionsLTR was significantly lower when a greater degree of iodized oil deposition occurred with a complete circumferential safety margin. In superselective TACE, the safety margin of the embolized areas using intraprocedural CBCT affected LTR in HCC patients.

Effect of treatment support on preventing local recurrence of hepatocellular carcinoma directly adjacent to the diaphragm.

Treatment support is anticipated to improve the results of radiofrequency ablation (RFA) treatment in cases in which visualization of tumors using the conventional B-mode is unclear. In the present study, the effectiveness of treatment support for RFA reducing the local recurrence rate of hepatocellular carcinoma (HCC) that are located directly adjacent to the diaphragm, and which are difficult to visualize with B-mode ultrasound imaging, was investigated. A total of 103 HCC tumors measuring <5 cm, which were located abutting the diaphragm, and which were difficult to visualize using the B-mode, were treated using RFA. Thirty-three of those HCC tumors were treated using RFA without treatment support, whereas the remaining 70 HCC tumors were treated using RFA with treatment support, including artificial pleural effusion, contrast-enhanced ultrasonography (CEUS) with the contrasting agent, Sonazoid™, and fusion imaging, either alone or in combination to improve the visualization of the tumors. The rate of local recurrence, and factors affecting local recurrence, were analyzed. Local recurrences were confirmed in 17 of the 103 nodules (16.50%). The overall rate of local recurrence was 13.1% at 6 months, and 20.2% at 12 months. The rate of local recurrence using RFA with artificial pleural effusion was significantly lower compared with those cases of HCC tumors treated without artificial pleural effusion (P=0.008). Similarly, the rate of local recurrence for CEUS RFA with Sonazoid™ was significantly lower compared with those cases of HCC tumors treated without Sonazoid™ (P=0.00081). In a multivariate analysis, CEUS RFA with Sonazoid™ and artificial pleural effusion contributed to the decrease in the rate of local recurrence (hazard ratios, 0.075 and 0.143, respectively). Based on these results, it is possible to conclude that CEUS with Sonazoid™ as a treatment support was the most effective method for reducing the rate of local recurrences abutting the diaphragm that are difficult to visualize using B-mode ultrasonography.

One‐month apparent diffusion coefficient correlates with response to radiofrequency ablation of hepatocellular carcinoma

To assess whether apparent diffusion coefficient (ADC) values at 1 and 3 months after radiofrequency ablation (RFA) may be associated with a favorable response to therapy for hepatocellular carcinoma (HCC) and liver metastases. Fifty-nine patients with HCC (n = 35) or liver metastases (n = 24) who underwent 1.5T diffusion-weighted magnetic resonance imaging (DWMRI) at 1 and 3 months post-RFA were included. ADC values of patients with local tumor recurrence were compared to those without local recurrence. A subgroup analysis was performed for HCC and metastases. Thirty-eight HCC and 27 metastases were evaluated. The ADC value of HCC at 1 month after RFA was lower in recurrent tumors (0.957 ± 0.229 [SD] × 10-3 mm2 ) compared to tumors with complete response (1.414 ± 0.322 [SD] × 10-3 mm2 /s, P = 0.006). At multivariate analysis, ADC at 1 month was the single independent variable associated with recurrence for HCC (area under the receiver operating characteristic curve = 0.860). No significant association was observed for liver metastases (P = 0.089). A low ADC value at 1 month after RFA is associated with an early local recurrence of HCC. This study does not confirm that such association exists for hepatic metastases. 3 Technical Efficacy: Stage 2 J. MAGN. RESON. IMAGING 2017;45:1648-1658.

Heat Stress-Induced PI3K/mTORC2-Dependent AKT Signaling Is a Central Mediator of Hepatocellular Carcinoma Survival to Thermal Ablation Induced Heat Stress.

Thermal ablative therapies are important treatment options in the multidisciplinary care of patients with hepatocellular carcinoma (HCC), but lesions larger than 2–3 cm are plagued with high local recurrence rates and overall survival of these patients remains poor. Currently no adjuvant therapies exist to prevent local HCC recurrence in patients undergoing thermal ablation. The molecular mechanisms mediating HCC resistance to thermal ablation induced heat stress and local recurrence remain unclear. Here we demonstrate that the HCC cells with a poor prognostic hepatic stem cell subtype (Subtype HS) are more resistant to heat stress than HCC cells with a better prognostic hepatocyte subtype (Subtype HC). Moreover, sublethal heat stress rapidly induces phosphoinositide 3-kinase (PI3K)/mammalian target of rapamycin (mTOR) dependent-protein kinase B (AKT) survival signaling in HCC cells in vitro and at the tumor ablation margin in vivo. Conversely, inhibition of PI3K/mTOR complex 2 (mTORC2)-dependent AKT phosphorylation or direct inhibition of AKT function both enhance HCC cell killing and decrease HCC cell survival to sublethal heat stress in both poor and better prognostic HCC subtypes while mTOR complex 1 (mTORC1)-inhibition has no impact. Finally, we showed that AKT isoforms 1, 2 and 3 are differentially upregulated in primary human HCCs and that overexpression of AKT correlates with worse tumor biology and pathologic features (AKT3) and prognosis (AKT1). Together these findings define a novel molecular mechanism whereby heat stress induces PI3K/mTORC2-dependent AKT survival signaling in HCC cells and provide a mechanistic rationale for adjuvant AKT inhibition in combination with thermal ablation as a strategy to enhance HCC cell killing and prevent local recurrence, particularly at the ablation margin.

Epithelial–mesenchymal transition-related genes are linked to aggressive local recurrence of hepatocellular carcinoma after radiofrequency ablation

We reported that poor prognoses of hepatocellular carcinoma (HCC) patients after radiofrequency ablation (RFA) are owing to up-regulation of expression of hypoxia-inducible factor-1 and epithelial cell adhesion molecule. We investigated aggressive progression in residual liver tumors (RLTs) after RFA to focus on expression of epithelial–mesenchymal transition (EMT)-related genes and miRNAs. Ten patients with recurrent HCC post-RFA who underwent hepatectomy (RFA group) and 78 patients with HCC without prior RFA (non-RFA group) were enrolled. We examined expression of transforming growth factor (TGF)-β, Twist, vimentin, and Snail-1 mRNAs in tumor tissues, and expression of miR-34a and miR-200c. Expression of TGF-β, Twist and Snail-1 in the RFA group was significantly higher than that in the non-RFA group (P < 0.05); vimentin expression in the RFA group was higher than that in the non-RFA group (P = 0.07). Expression of miR-200c and miR-34a in the RFA group was significantly lower than that in the non-RFA group (miR-200c: P = 0.04; miR-34a: P < 0.01). Increased expression of EMT markers through down-regulation of miRNA expression in RLTs after RFA may be related to poor prognoses of HCC patients with aggressive local recurrence after RFA.

Recurrent feature after radiofrequency ablation therapy for hepatocelluar carcinoma: Risk of increasing malignant behavior.

213 Background: Radiofrequency ablation (RFA) is a widely used therapy for hepatocellular carcinoma (HCC). However, several reports demonstrated the aggressive local recurrence after RFA, suggesting the induction of further malignant transformation of HCC. Methods: Eighty-eight patients with HCC who underwent hepatic resection were included in this study. Hepatectomy was indicated for local recurrence of HCC after RFA (n = 10, RFA group) and for the HCC without prior RFA (n = 78, non-RFA group). Clinicopathological data and patient’s prognosis after hepatectomy were compared between the two groups. Expression levels of HIF-1, EpCAM, CD44, VEGF and EMT markers mRNA in the tumor tissues were examined. We also investigated miR-34a and miR-200c expressions in the two groups. Results: RFA group showed higher frequency of portal vein invasion and less tumor differentiation compared with the non-RFA group (p &lt; 0.05). Overall and disease-free survival rates in the RFA group were significantly worse than those in the non-RFA group (p &lt; 0.05). HIF-1 and EpCAM mRNA expression levels in the RFA group were significantly higher than those in the non-RFA group (p &lt; 0.05). The expressions of TGF-β, Twist and Snail-1 in RFA group were significantly higher than those in non-RFA group (P &lt; 0.05), and Vimentin expression in RFA group tended to be higher than that in non-RFA group (P = 0.07). Regarding to miRNA, miR-200c and miR-34a expressions in RFA group were significantly lower than those in non-RFA group (miR-200c: P = 0.04, miR-34a: P &lt; 0.01). Conclusions: Recurrent HCC after RFA showed the highly malignant potential through the expressions of HIF-1, cancer stem cell and EMT markers.

Radiofrequency ablation-increased CXCL10 is associated with earlier recurrence of hepatocellular carcinoma by promoting stemness.

Radiofrequency ablation (RFA) represents a valuable choice in hepatocellular carcinoma (HCC); however, local recurrence of HCC is common after RFA. Here, 20 primary HCC patients treated by RFA were enrolled. Before (termed 0d) and after RFA treatment for 1 and 7days (termed 1d and 7d, respectively), plasma and noncancerous tissue were collected. ELISA assay showed that plasma C-X-C motif chemokine 10 (CXCL10) was increased in ten patients (type I patients) but decreased in the other 10 patients (type II patients). The mean interval for HCC recurrence in type I patients was less than the mean interval in type II patients. Interestingly, a significant negative correlation between interval for HCC recurrence and fold change of plasma CXCL10 (1d/0d or 7d/0d) was identified, suggesting that RFA-induced CXCL10 is associated with earlier HCC recurrence. Immunofluorescence assay showed that the receptor of CXCL10, chemokine (C-X-C motif) receptor 3 (CXCR3), was significantly increased in type I, but not type II, patients after RFA. In vitro assay demonstrated that CXCL10 stimulus increased the rate of CD133(+) cancer stem cells (CSCs) in HepG2 cells by binding to CXCR3 and then inducing c-Myc expression. Many studies have reported that induction of CD133(+) CSCs contributes to HCC recurrence. Thus, CXCL10-increased CD133(+) CSCs by activating CXCR3/c-Myc pathway might accelerate HCC recurrence after RFA. These data might have potential implications for HCC therapy.

Salvage treatment for local recurrence of hepatocellular carcinoma after local ablation therapy

Local recurrence of hepatocellular carcinoma (HCC) after local ablation therapy (LAT) is a serious problem; however, the optimal treatment strategy remains unclear. A retrospective analysis was conducted of 50 patients with local recurrence of HCC after LAT that underwent either salvage hepatectomy (n = 23) or radiofrequency ablation (RFA; n = 27). Their background characteristics, intraoperative data, and postoperative short- and long-term outcome were analyzed. The RFA group was found to be significantly associated with an impaired liver functional reserve, smaller number and size of tumors. The hepatectomy group showed a significantly longer operation time, greater intraoperative blood loss and more frequent red blood cell transfusion. In-hospital stay mortality and morbidity rate did not significantly differ. The disease-free and overall survival showed no significant difference between the groups. Although local recurrence after salvage treatment was found in zero (0%) for hepatectomy and in seven (25.9%) for RFA, that did not affect the overall survival. The tumor size at the prior LAT was identified as an independent prognostic factor for disease-free survival and serum albumin for overall survival. Whereas salvage hepatectomy for local recurrent HCC is recommended for selected patients in terms of its good local control effect, salvage RFA is also acceptable because it is less invasive and also has a reasonable long-term outcome.

Expressions of Hypoxia-Inducible Factor-1 and Epithelial Cell Adhesion Molecule are Linked with Aggressive Local Recurrence of Hepatocellular Carcinoma After Radiofrequency Ablation Therapy

Radiofrequency ablation (RFA) is a widely used therapy for hepatocellular carcinoma (HCC). Several reports have demonstrated the aggressive local recurrence of HCC after RFA, suggesting that induction of further malignant transformation of HCC has occurred. Eighty-eight (88) patients with HCC who underwent hepatic resection were included in this study. Hepatectomy was indicated for local recurrence of HCC after RFA (n = 10, RFA group) and for HCC without prior RFA (n = 78, non-RFA group). Clinicopathological data and the patient's prognosis after hepatectomy were compared between the two groups. Expression levels of hypoxia-inducible factor-1 (HIF-1), epithelial cell adhesion molecule (EpCAM), CD44, and vascular endothelial growth factor messenger RNA (mRNA) in the tumor tissues were also examined. The RFA group showed higher frequency of portal vein invasion and less tumor differentiation compared with the non-RFA group (p < 0.05). Overall and disease-free survival rates in the RFA group were significantly worse than those in the non-RFA group (p < 0.05). HIF-1 and EpCAM mRNA expression levels in the RFA group were significantly higher than those in the non-RFA group (p < 0.05). These results suggest that local HCC recurrence after RFA shows an aggressive tumor phenotype and poor prognosis through the enhanced expressions of HIF-1 and EpCAM in the residual HCC tumors after insufficient or sub-lethal treatment by RFA.