Bile acids,gut microbiota,and therapeutic insights in hepatocellular carcinoma

Hepatocellular carcinoma(HCC)is a prevalent and aggressive liver malignancy.The interplay between bile acids(BAs)and the gut microbiota has emerged as a critical factor in HCC development and progression.Under normal conditions,BA metabolism is tightly regulated through a bidirectional interplay between gut microorganisms and BAs.The gut microbiota plays a critical role in BA metabolism,and BAs are endogenous signaling molecules that help maintain liver and intestinal homeostasis.Of note,dysbiotic changes in the gut microbiota during pathogenesis and cancer development can disrupt BA homeostasis,thereby leading to liver inflammation and fibrosis,and ultimately contributing to HCC development.Therefore,understanding the intricate interplay between BAs and the gut microbiota is crucial for elucidating the mechanisms underlying hepatocarcinogenesis.In this review,we comprehensively explore the roles and functions of BA metabolism,with a focus on the interactions between BAs and gut microorganisms in HCC.Additionally,therapeutic strategies targeting BA metabolism and the gut microbiota are discussed,including the use of BA agonists/antagonists,probiotic/prebiotic and dietary interventions,fecal microbiota transplantation,and engineered bacteria.In summary,understanding the complex BA-microbiota crosstalk can provide valuable insights into HCC development and facilitate the development of innovative therapeutic approaches for liver malignancy.

Fatty acid binding protein 5 is a novel therapeutic target for hepatocellular carcinoma

BACKGROUND Hepatocellular carcinoma(HCC)is an aggressive subtype of liver cancer and is one of the most common cancers with high mortality worldwide.Reprogrammed lipid metabolism plays crucial roles in HCC cancer cell survival,growth,and evolution.Emerging evidence suggests the importance of fatty acid binding proteins(FABPs)in contribution to cancer progression and metastasis;however,how these FABPs are dysregulated in cancer cells,especially in HCC,and the roles of FABPs in cancer progression have not been well defined.AIM To understand the genetic alterations and expression of FABPs and their associated cancer hallmarks and oncogenes in contributing to cancer malignancies.METHODS We used The Cancer Genome Atlas datasets of pan cancer and liver hepatocellular carcinoma(LIHC)as well as patient cohorts with other cancer types in this study.We investigated genetic alterations of FABPs in various cancer types.mRNA expression was used to determine if FABPs are abnormally expressed in tumor tissues compared to non-tumor controls and to investigate whether their expression correlates with patient clinical outcome,enriched cancer hallmarks and oncogenes previously reported for patients with HCC.We determined the protein levels of FABP5 and its correlated genes in two HCC cell lines and assessed the potential of FABP5 inhibition in treating HCC cells.RESULTS We discovered that a gene cluster including five FABP family members(FABP4,FABP5,FABP8,FABP9 and FABP12)is frequently co-amplified in cancer.Amplification,in fact,is the most common genetic alteration for FABPs,leading to overexpression of FABPs.FABP5 showed the greatest differential mRNA expression comparing tumor with non-tumor tissues.High FABP5 expression correlates well with worse patient outcomes(P<0.05).FABP5 expression highly correlates with enrichment of G2M checkpoint(r=0.33,P=1.1e-10),TP53 signaling pathway(r=0.22,P=1.7e-5)and many genes in the gene sets such as CDK1(r=0.56,P=0),CDK4(r=0.49,P=0),and TP53(r=0.22,P=1.6e-5).Furthermore,FABP5 also correlates well with two co-expressed oncogenes PLK1 and BIRC5 in pan cancer especially in LIHC patients(r=0.58,P=0;r=0.58,P=0;respectively).FABP5high Huh7 cells also expressed higher protein levels of p53,BIRC5,CDK1,CDK2,and CDK4 than FABP5low HepG2 cells.FABP5 inhibition more potently inhibited the tumor cell growth in Huh7 cells than in HepG2 cells.CONCLUSION We discovered that FABP5 gene is frequently amplified in cancer,especially in HCC,leading to its significant elevated expression in HCC.Its high expression correlates well with worse patient outcome,enriched cancer hallmarks and oncogenes in HCC.FABP5 inhibition impaired the cell viability of FABP5high Huh7 cells.All these support that FABP5 is a novel therapeutic target for treating FABP5high HCC.

Suberoylanilide hydroxamic acid upregulates reticulophagy receptor expression and promotes cell death in hepatocellular carcinoma cells

BACKGROUND Hepatocellular carcinoma(HCC)is a common clinical condition with a poor prognosis and few effective treatment options.Potent anticancer agents for treating HCC must be identified.Epigenetics plays an essential role in HCC tumorigenesis.Suberoylanilide hydroxamic acid(SAHA),the most common histone deacetylase inhibitor agent,triggers many forms of cell death in HCC.However,the underlying mechanism of action remains unclear.Family with sequence similarity 134 member B(FAM134B)-induced reticulophagy,a selective autophagic pathway,participates in the decision of cell fate and exhibits anticancer activity.This study focused on the relationship between FAM134B-induced reticulophagy and SAHA-mediated cell death.AIM To elucidate potential roles and underlying molecular mechanisms of reticulophagy in SAHA-induced HCC cell death.METHODS The viability,apoptosis,cell cycle,migration,and invasion of SAHA-treated Huh7 and MHCC97L cells were measured.Proteins related to the reticulophagy pathway,mitochondria-endoplasmic reticulum(ER)contact sites,intrinsic mitochondrial apoptosis,and histone acetylation were quantified using western blotting.ER and lysosome colocalization,and mitochondrial Ca^(2+)levels were characterized via confocal microscopy.The level of cell death was evaluated through Hoechst 33342 staining and propidium iodide colocalization.Chromatin immunoprecipitation was used to verify histone H4 lysine-16 acetylation in the FAM134B promoter region.RESULTS After SAHA treatment,the proliferation of Huh7 and MHCC97L cells was significantly inhibited,and the migration and invasion abilities were greatly blocked in vitro.This promoted apoptosis and caused G1 phase cells to increase in a concentration-dependent manner.Following treatment with SAHA,ER-phagy was activated,thereby triggering autophagy-mediated cell death of HCC cells in vitro.Western blotting and chromatin immunoprecipitation assays confirmed that SAHA regulated FAM134B expression by enhancing the histone H4 lysine-16 acetylation in the FAM134B promoter region.Further,SAHA disturbed the Ca^(2+)homeostasis and upregulated the level of autocrine motility factor receptor and proteins related to mitochondria-endoplasmic reticulum contact sites in HCC cells.Additionally,SAHA decreased the mitochondrial membrane potential levels,thereby accelerating the activation of the reticulophagy-mediated mitochondrial apoptosis pathway and promoting HCC cell death in vitro.CONCLUSION SAHA stimulates FAM134B-mediated ER-phagy to synergistically enhance the mitochondrial apoptotic pathway,thereby enhancing HCC cell death.

Zoledronic acid inhibits growth of hepatocellular carcinoma cells in vitro and in vivo

Background Growing preclinical evidence shows that zoledronic acid （ZOL） exhibits direct antitumor activity in various cancer cell lines. However, the cytotoxic effects of ZOL on human hepatocellular carcinoma （HCC） cells have not been established. In the present study, we investigated the effect of ZOL on HCC both in vitro and in vivo. Methods Cytotoxicity and cell cycles were assessed with Sulforhodamine B colorimetric assay and flow cytometry. Expression levels of cell cycle phase-linked proteins were examined. The effect of ZOL on HCC in vivo was explored based on H22-subcutaneous injection （s.c.） and H22-intraperitoneal injection （i.p.） mice model. Results ZOL inhibited the growth of SK-HEP-1 and H22 cells and induced S-phase arrest through downregulating cdc2 protein and upregulating cyclin A. It inhibited the growth of s.c tumors, and increased the survival of both H22-s.c. and H22-i.p. mice in vivo. Conclusion ZOL inhibits qrowth of HCC cells in vitro and in vivo.

Gadoxetic acid-enhanced magnetic resonance imaging can predict the pathologic stage of solitary hepatocellular carcinoma

BACKGROUND Although important for determining long-term outcome, pathologic stage of hepatocellular carcinoma (HCC) is difficult to predict before surgery. Current state-of-the-art magnetic resonance imaging (MRI) using gadoxetic acid provides many imaging features that could potentially be used to classify single HCC as pT1 or pT2. AIM To determine which gadoxetic acid-enhanced MRI (EOB-MRI) findings predict pathologic stage T2 in patients with solitary HCC (cT1). METHODS Pre-operative EOB-MRI findings were reviewed in a retrospective cohort of patients with solitary HCC. The following imaging features were examined: Hyperintensity in unenhanced T2-weighted images, hypointensity in unenhanced T1-weighted images, arterial enhancement, corona enhancement, washout appearance, capsular appearance, hypointensity in the tumor tissue during the hepatobiliary (HB) phase, peritumoral hypointensity in the HB phase, hypointense rim in the HB phase, intratumoral fat, hyperintensity on diffusionweighted imaging, hypointensity on apparent diffusion coefficient map, mosaic appearance, nodule-in-nodule appearance, and the margin (smooth or irregular). Surgical pathology was used as the reference method for tumor staging. Univariate and multivariate analyses were performed to identify predictors of microvascular invasion or satellite nodules. RESULTS There were 39 (34.2%;39 of 114) and 75 (65.8%;75 of 114) pathological stage T2 and T1 HCCs, respectively. Large tumor size (≥ 2.3 cm) and two MRI findings, i.e., corona enhancement [odds ratio = 2.67;95% confidence interval: 1.101-6.480] and peritumoral hypointensity in HB phase images (odds ratio = 2.203;95% confidence interval: 0.961-5.049) were associated with high risk of pT2 HCC. The positive likelihood ratio was 6.25 (95% confidence interval: 1.788-21.845), and sensitivity of EOB-MRI for detecting pT2 HCC was 86.2% when two or three of these MRI features were present. Small tumor size and hypointense rim in the HB phase were regarded as benign features. Small HCCs with hypointense rim but not associated with aggressive features were mostly pT1 lesions (specificity, 100%). CONCLUSION Imaging features on EOB-MRI could potentially be used to predict the pathologic stage of solitary HCC (cT1) as pT1 or pT2.

Hydrochloric acid-enhanced radiofrequency ablation for treating a large hepatocellular carcinoma with spontaneous rapture:a case report

Background:A ruptured hepatocellular carcinoma(HCC)is often fatal.In addition to surgery and transarterial embo?lization,radiofrequency ablation(RFA)might be another option for treating a ruptured HCC.Unfortunately,conven?tional RFA has a limited ablation zone;as such,it is rarely used to treat ruptured tumors.Case presentation:This case was a 60?year?old man who had a large,ruptured HCC in which hydrochloric acid(HCl)?enhanced RFA successfully controlled the bleeding and made the tumor completely necrotic.Conclusion:Considering the effectiveness of HCl?enhanced RFA in achieving hemostasis and tumor ablation,it might be a new option for treating large,ruptured HCCs.

Added value of hepatobiliary phase gadoxetic acid-enhanced MRI for diagnosing hepatocellular carcinoma in high-risk patients

AIM:To determine the added value of hepatobiliary phase(HBP)gadoxetic acid-enhanced magnetic resonance imaging(MRI)in evaluating hepatic nodules in high-risk patients.METHODS:The institutional review board approved this retrospective study and waived the requirement for informed consent.This study included 100 patients at high risk for hepatocellular carcinoma(HCC)and 105hepatic nodules that were larger than 1 cm.A blind review of two MR image sets was performed in a random order:set 1,unenhanced(T1-and T2-weighted)and dynamic images;and set 2,unenhanced,dynamic20-min and HBP images.The diagnostic accuracy,sensitivity,specificity,positive predictive value(PPV),and negative predictive value(NPV)were compared for the two image sets.Univariate and multivariate analyses were performed on the MR characteristics utilized to diagnose HCC.RESULTS:A total of 105 hepatic nodules were identified in 100 patients.Fifty-nine nodules were confirmed to be HCC.The diameter of the 59 HCCs ranged from1 to 12 cm(mean:1.9 cm).The remaining 46 nodules were benign(28 were of hepatocyte origin,nine were hepatic cysts,seven were hemangiomas,one was chronic inflammation,and one was focal fat infiltration).The diagnostic accuracy significantly increased with the addition of HBP images,from 88.7%in set 1to 95.5%in set 2(P=0.002).In set 1 vs set 2,the sensitivity and NPV increased from 79.7%to 93.2%and from 78.9%to 91.8%,respectively,whereas the specificity and PPV were not significantly different.The hypointensity on the HBP images was the most sensitive(93.2%),and typical arterial enhancement followed by washout was the most specific(97.8%).The multivariate analysis revealed that typical arterial enhancement followed by washout,hyperintensity on T2-weighted images,and hypointensity on HBP images were statistically significant MRI findings that could diagnose HCC(P<0.05).CONCLUSION:The addition of HBP gadoxetic acidenhanced MRI statistically improved the diagnostic accuracy in HCCs larger than 1 cm.Typical arterial enhancement followed by washout and hypointensity on HBP images are useful for diagnosing HCC.

Omega-3 fatty acid improves the clinical outcome of hepatectomized patients with hepatitis B virus(HBV)-associated hepatocellular carcinoma

Omega-3 fatty acid supplemented total parenteral nutrition improves the clinical outcome of patients undergoing certain operations; however, its benefits for patients with hepatitis type B virus (HBV)-associated hepatocellular carcinoma (HCC) who have undergone hepatectomy are still not clear. The aim of this study was to evaluate the effect of omega-3 fatty acid supplemented total parenteral nutrition on the clinical outcome of patients with HBV- associated HCC who underwent hepatectomy at our institution. A total of 63 patients with HBV-associated HCC who underwent hepatectomy were included in this study. These patients were randomly assigned to receive stand- ard total parenteral nutrition (the control group, n = 31) or omega-3 fatty acid supplemented total parenteral nutri- tion (the omega-3 fatty acid group, n = 32) for at least 5 d. The study endpoints were the occurrence of infection- related complications, recovery of liver function and length of hospital stay. The results showed that the omega-3 fatty acid group had a lower infection rate (omega-3 fatty acid, 19.4% vs control, 43.8%, P 〈 0.05), a better liver function after hepatectomy: alanine transaminase (omega-3 fatty acid, 48.23__+ 18.48 U/L vs control, 73.34 q-40.60 U/L, P 〈 0.01), aspartate transaminase (omega-3 fatty acid, 35.77_ 14.56 U/L vs control, 50.53 4-24.62 U/L, P 〈 0.01), total bilirubin (omega-3 fatty acid, 24.29___7.40 mmol/L vs control, 28. 374-8.06 mmol/L, P 〈 0.05) and a shorter length of hospital stay (omega-3 fatty acid, 12.71 ___2.58 d vs control, 15.91 ___3.23 d, P 〈 0.01). The serum contents of IL-6 (omega-3 fatty acid, 23.98___5.63 pg/mL vs control, 35.55___7.5 pg/mL, P 〈 0.01) and TNF-a (ome- ga-3 fatty acid, 4.43___1.22 pg/mL vs control, 5.96___1.58 pg/mL, P 〈 0.01) after hepatectomy were significantly lower in the omega-3 fatty acid group than those of the control group. In conclusion, administration of omega-3 fatty acid may reduce infection rate and improve liver function recovery in HBV-associated HCC patients after hepatectomy. This improvement is associated with suppressed production of proinflammatory cytokines in these patients.

Blood exosomal micro ribonucleic acid profiling reveals the complexity of hepatocellular carcinoma and identifies potential biomarkers for differential diagnosis

BACKGROUND Hepatocellular carcinoma(HCC)is one of the leading causes of cancer-related deaths worldwide,but there is a shortage of effective biomarkers for its diagnosis.AIM To explore blood exosomal micro ribonucleic acids(miRNAs)as potential biomarkers for HCC diagnosis.RESULTS The principal component analysis suggested that daily alcohol consumption could alter the blood exosomal miRNA profiles of hepatitis B virus positive non-HCC patients through miR-3168 and miR-223-3p.The miRNA profiles also revealed the tumor stages of HCC patients.High expression of miR-455-5p and miR-30c-5p,which significantly correlated with better overall survival in tumor tissues,could also be detected in blood exosomes.Two pairs of miRNAs(miR-584-5p/miR-106-3p and miR-628-3p/miR-941)showed a 94.1%sensitivity and 68.4%specificity to differentiate HCC patients from non-HCC patients.The specificity of the combination was substantially influenced by alcohol consumption habits.CONCLUSION This study suggested that blood exosomal miRNAs can be used as new noninvasive diagnostic tools for HCC.However,their accuracy could be affected by tumor stage and alcohol consumption habits.

Hydrochloric acid enhanced radiofrequency ablation for treatment of large hepatocellular carcinoma in the caudate lobe:Report of three cases

BACKGROUND To report on the use of percutaneous hydrochloric acid(HCl) enhanced radiofrequency ablation(HRFA) for the treatment of large(maximum diameter ≥5 cm) hepatocellular carcinoma(HCC) in the caudate lobe.CASE SUMMARY Between August 2013 and June 2016, three patients with a large HCC(maximum diameter: 5.0, 5.7, and 8.1 cm) in the caudate lobe were treated by transarterial chemoembolization followed by computer tomography(CT) guided RFA using a monopolar perfusion RF electrode, which was enhanced by local infusion of 10%HCl at 0.2 mL/min(total volume, 3 to 12 mL). The output power of HRFA reached 100 W, and the average ablation time was 39 min(range, 15 to 60 min).Two patients each underwent one session of HRFA and one patient two sessions.After treatment, CT/magnetic resonance imaging showed that all the three lesions were completely ablated. There was no major complication. Two patients had asymptomatic bile duct dilatation. One patient died of tongue cancer 24 mo after ablation. The remaining two patients were alive and no area of enhancement is detected in the caudate lobe at 28 and 60 mo after ablation, respectively.CONCLUSION Percutaneous CT-guided HRFA is safe and efficacious in treating large HCC in the caudate lobe.

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

Objective:Local 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.Methods:The proportions of CD133+and epithelial cell adhesion molecule(Ep CAM);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 Ep CAM+TICs and expression of stem cell-related genes,was evaluated in vitro and in vivo.Finally,the effect of ATRA on the tumor growth and the proportion of TICs was evaluated.Results:In clinical data,a higher proportion of CD133+and Ep CAM+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 Ep CAM+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 PI3 K/AKT pathway.Combination treatment with ATRA resulted in longer survival outcomes in mouse xenografts than RFA alone.Conclusions:ATRA,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.

Diagnostic efficacy of gadoxetic acid-enhanced MRI for hepatocellular carcinoma and dysplastic nodule

AIM:To evaluate the relationship between the signal intensity of hepatobiliary phase images on gadoxetic acid-enhanced magnetic resonance imaging(MRI)and histological grade.METHODS:Fifty-nine patients with 82 hepatocellular lesions were evaluated retrospectively.Hepatobiliary phase images on gadoxetic acid-enhanced MRI were classified into 3 groups:low,iso or high.Angiographyassisted computed tomography(CT)findings were also classified into 3 groups:CT during arterial portography,and CT hepatic arteriography:A:iso,iso or low;B:slightly low,iso or low;;and C:low,high.We correlated angiography-assisted CT,hepatobiliary phase findings during gadoxetic acid-enhanced MRI and histological grades.Furthermore,correlations between MRI findings and histological grade for each hemodynamic pattern were performed.Correlations among radiologicaland pathological findings were statistically evaluated using the chi-square test and Fisher's exact test.RESULTS:There was a significant correlation between histological grade and hemodynamic pattern(P<0.05).There was a significant correlation between histological grade and signal intensity in the hepatobiliary phase(P<0.05)in group A lesions.There was no significant correlation between histological grade and signal intensity in the hepatobiliary phase in group B or C lesions(P>0.05).CONCLUSION:Signal intensity in the hepatobiliary phase correlated with histological grade in the lesions that maintained portal blood flow,but did not correlate in lesions that showed decreased or defective portal blood flow.

Liquid biopsy in patients with hepatocellular carcinoma: Circulating tumor cells and cell-free nucleic acids

Hepatocellular carcinoma(HCC), with its high incidence and mortality rate, is one of the most common malignant tumors. Despite recent development of a diagnostic and treatment method, the prognosis of HCC remains poor. Therefore, to provide optimal treatment for each patient with HCC, more precise and effective biomarkers are urgently needed which could facilitate a more detailed individualized decision-making during HCC treatment, including the following; risk assessment, early cancer detection, prediction of treatment or prognostic outcome. In the blood of cancer patients, accumulating evidence about circulating tumor cells and cell-free nucleic acids has suggested their potent clinical utilities as novel biomarker. This concept, so-called "liquid biopsy" is widely known as an alternative approach to cancer tissue biopsy. This method might facilitate a more sensitive diagnosis and better decision-making by obtaining genetic and epigenetic aberrations that are closely associated with cancer initiation and progression. In this article, we review recent developments based on the available literature on both circulating tumor cells and cell-free nucleic acids in cancer patients, especially focusing on Hepatocellular carcinoma.

Branched-chain amino acids to tyrosine ratio value as a potential prognostic factor for hepatocellular carcinoma

The prognosis of hepatocellular carcinoma(HCC) depends on tumor extension as well as hepatic function.Hepatic functional reserve is recognized as a factor affecting survival in the treatment of HCC;the ChildPugh classif ication system is the most extensively used method for assessing hepatic functional reserve in patients with chronic liver disease,using serum albumin level to achieve accurate assessment of the status of protein metabolism.However,insuff icient attention has been given to the status of amino acid(AA) metabolism in chronic liver disease and HCC.Fischer's ratio is the molar ratio of branched-chain AAs(BCAAs:leucine,valine,isoleucine) to aromatic AAs(phenylalanine,tyrosine) and is important for assessing liver metabolism,hepatic functional reserve and the severity of liver dysfunction.Although this ratio is diff icult to determine in clinical situations,BCAAs/tyrosine molar concentration ratio(BTR) has been proposed as a simpler substitute.BTR correlates with various liver function examinations,including markers of hepatic f ibrosis,hepatic blood flow and hepatocyte function,and can thus be considered as reflecting the degree of hepatic impairment.This manuscript examines the literature to clarify whether BTR can serve as a prognostic factor for treatment of HCC.

Gamma-aminobutyric acid promotes human hepatocellular carcinoma growth through overexpressed gamma-aminobutyric acid A receptor α3 subunit

AIM:To investigate the expression pattern of gamma-aminobutyric acid A(GABAA) receptors in hepatocellular carcinoma(HCC) and indicate the relationship among gamma-aminobutyric acid(GABA),gamma-aminobutyric acid A receptor α3 subunit(GABRA3) and HCC.METHODS:HCC cell line Chang,HepG2,normal liver cell line L-02 and 8 samples of HCC tissues and paired non-cancerous tissues were analyzed with semiquantitative polymerase chain reaction(PCR) for the expression of GABAA receptors.HepG2 cells were treated with gamma-aminobutyric acid(GABA) at serial concentrations(0,1,10,20,40 and 60 μmol/L),and their proliferating abilities were analyzed with the 3-(4,5-methylthiazol-2-yl)-2,5-diphenyltetrazolium bromide(MTT) assay,cell doubling time test,colon formation assay,cell cycle analysis and tumor planted in nude mice.Small interfering RNA was used for knocking down the endogenous GABRA3 in HepG2.Proliferating abilities of these cells treated with or without GABA were analyzed.RESULTS:We identified the overexpression of GABRA3 in HCC cells.Knockdown of endogenous GABRA3 expression in HepG2 attenuated HCC cell growth,suggesting its role in HCC cell viability.We determined the in vitro and in vivo effect of GABA in the proliferation of GABRA3-positive cell lines,and found that GABA increased HCC growth in a dose-dependent manner.Notably,the addition of GABA into the cell culture medium promoted the proliferation of GABRA3-expressing HepG2 cells,but not GABRA3-knockdown HepG2 cells.This means that GABA stimulates HepG2 cell growth through GABRA3.CONCLUSION:GABA and GABRA3 play important roles in HCC development and progression and can be a promising molecular target for the development of new diagnostic and therapeutic strategies for HCC.

Branched-chain amino acid treatment before transcatheter arterial chemoembolization for hepatocellular carcinoma

AIM:To examine the significance of branched-chain amino acid (BCAA) treatment before transcatheter arterial chemoembolization (TACE) for hepatocellular carcinoma (HCC).METHODS:This study included 99 patients who underwent TACE therapy for HCC at our hospital and were followed up without treatment for at least 6 mo between January 2004 and January 2010.They were divided into 2 groups:those receiving BCAA granules (n=40) or regular diet (n=59,control).Data obtained were retrospectively analyzed (prior to TACE,and 1 wk,1,3,and 6 mo after TACE) in terms of nutritional condition and clinical laboratory parameters (serum albumin level and Child-Pugh score),both of which are determinants of hepatic functional reserve.RESULTS:The BCAA group comprised 27 males and 13 females with a mean age of 69.9 ± 8.8 years.The patients of the BCAA group were classified as follows:Child-Pugh A/B/C in 22/15/3 patients,and Stage Ⅱ/Ⅲ /ⅣA HCC in 12/23/5 patients,respectively.The controlgroup comprised 32 males and 27 females with a mean age of 73.2 ± 10.1 years.In the control group,9 patients had chronic hepatitis,Child-Pugh A/B/C in 39/10/1 patients,and StageⅠ/Ⅱ/Ⅲ/ⅣA HCC in 1/11/35/12 patients,respectively.Overall,both serum albumin level and Child-Pugh score improved significantly in the BCAA group as compared with the control 3 and 6 mo after TACE (P < 0.05).Further analysis was performed by the following categorization:(1) child-Pugh classification;(2) liver cirrhosis subgroup with a serum albumin level > 3.5 g/dL;and (3) epirubicin dose.A similar trend indicating a significant improvement of all variables in the BCAA group was noted (P < 0.05).CONCLUSION:Treatment with BCAA granules in patients who have undergone TACE for HCC is considered useful to maintain their hepatic functional reserve.

Effect of branched-chain amino acids in patients receiving intervention for hepatocellular carcinoma

AIM:To investigate the usefulness of branched-chain amino acids(BCAA)before transarterial chemoembolization(TACE)or radiofrequency ablation(RFA).METHODS:We investigated the usefulness of preintervention with BCAAs by comparing patients treated with BCAAs at 12.45 g/d orally for at least 2 wk before TACE or RFA and those not receiving such pretreatment.A total of 270 patients with hepatocellular carcinoma complicated by cirrhosis were included in the study.Mean changes from baseline(Δ)in serum albumin(Alb),C-reactive protein(CRP),and transaminase levels,as well as peak body temperature were also determined and compared at days 2,5,and 10 after the start of TACE or RFA.RESULTS:In patients who underwent TACE or RFA,BCAA pre-intervention significantly suppressed the development of post-intervention hypoalbuminemia and reduced inflammatory reactions during the subsequent clinical course.After TACE,theΔAlb peaked on day 2,remained constantly lower in BCAA-treated patients,compared to the control group,and was-0.13±0.42g/dL in BCAA-treated patients and-0.33±0.51 g/dL in untreated patients on day 10.TheΔCRP was also significantly lower in BCAA-treated patients on days2,5 and 10 after TACE.Like the trends noted after TACE,a similar tendency was noted as to theΔAlb andΔCRP after RFA.The changes in serum Alb level were inversely correlated with CRP changes;therefore,a possible involvement of the anti-inflammatory effect of BCAAs was inferred as a factor contributory to the suppression of decrease in serum Alb level.CONCLUSION:Pre-intervention with BCAAs may hasten the recovery of serum Alb level and mitigate postoperative complications in patients undergoing TACE or RFA.

Activity and safety of pegylated liposomal doxorubicin,5-fluorouracil and folinic acid in inoperable hepatocellular carcinoma:A phase Ⅱ study

AIM: To improve the results of New therapeutic strategies in hepatocellular carcinoma (HCC). We have conducted a phase Ⅱ study with pegylated liposomal doxorubicin (PLD), 5-fluorouracil (5FU) and folinic acid (FA). METHODS: Thirty-one patients with hystologically- confirmed, inoperable HCC, received combination chemotherapy with PLD 25 mg/mq on d 1, 5FU 1200 mg/mq in 48 h continuous infusion, and oral FA 30 mg on d 1 and 2 every 3 wk until disease progression or intolerable toxicity. RESULTS: The median age was 65 years (range 41-82) and 28 patients were hepatitis C virus seropositive (90%). The majority of patients were Child-Pugh Class B (55%). Two patients showed a partial response (PR), and 16 had stable disease (SD). With a median follow-up of 14 mo, the median time to progression of all evaluable patients was 4 mo (95% CI 1.7-7). Median overall survival was 9 mo (95% CI 3-24 mo). After 1 year, 9 of 18 PR/SD patients were alive. Chemotherapy was well tolerated. CONCLUSION: PLD/FU/FA combination seems capable of achieving durable stabilization of HCC. The manageable toxicity supports a role for combination with other anticancer agents.

Gadoxetic acid-enhanced magnetic resonance imaging for the detection of small hepatocellular carcinoma(≤ 2.0 cm) in patients with chronic liver disease:A meta-analysis

AIM To perform a meta-analysis assessing the value of gadoxetic acid-enhanced magnetic resonance imaging(Gd-EOB-MRI)in detecting small hepatocellular carcinoma(HCC)(≤2.0 cm)in patients with chronic liver disease.METHODS Databases,including MEDLINE and EMBASE,were searched for relevant original articles published from January 2008 to February 2015.Data were extracted,and summary estimates of diagnostic accuracy indexes such as sensitivity,specificity,diagnostic odds ratio,predictive value,and areas under summary receiver operating characteristic curve were obtained using a random-effects model,with further exploration employing meta-regression and subgroup analyses.RESULTS In 10 studies evaluating 768 patients,pooled perlesion sensitivity of Gd-EOB-DTPA was 91%(95%CI:83%-95%),with a specificity of 95%(95%CI:87%-98%).Overall positive likelihood ratio was 18.1(95%CI:6.6-49.4),for negative likelihood ratio(NLR)of 0.10(95%CI:0.05-0.19)and diagnostic odds ratio of182(95%CI:57-581).Subgroup analysis suggested that diagnostic performance of Gd-EOB-MRI for sub-centimeter HCC(≤1.0 cm)detection was low,with a sensitivity of69%(95%CI:59%-78%).In studies with both Gd-EOBMRI and diffusion-weighted imaging(DWI)performed,Gd-EOB-MRI/DWI combination was more sensitive than Gd-EOB-DTPA alone,whether for small lesions(86%vs77%)or sub-centimeter ones(80%vs 56%).CONCLUSION A limited number of small studies suggested that GdEOB-MRI has good diagnostic performance in the detection of small HCC(≤2.0 cm)among patients with chronic liver disease,but relatively lower performance for detection of sub-centimeter HCC(≤1.0 cm).Combination of Gd-EOB-MRI and DWI can improve the diagnostic sensitivity of MRI.

Relationship between Serum Hyaluronic Acid Level and Patient Prognosis in Hepatocellular Carcinoma Patients Who Underwent Hepatectomy

We examined the relationship between hyaluronic acid (HA) and tumor-related factors after hepatectomy in 158 patients with hepatocellular carcinoma (HCC) who underwent hepatectomy. We examined serum HA levels before hepatectomy by evaluating data for clinicopathological parameters, surgical records, postoperative complications and survival. The mean HA level was 190 ± 202 ng/ml. Patients were divided into 3 groups: group A had serum HA levels less than 50 ng/ml (normal range), group B had levels between 50 and 190 ng/ml, and group C had levels over 190 ng/ml. Group C had a higher rate of poor liver function compared to others. Multiple tumors were significantly more frequent in groups B and C compared to A. The grade of fibrosis and the inflammatory responses were positively correlated with the serum HA level. Postoperative long-term ascites was significantly more frequent in group C compared to others. Although the recurrence rate and the relapse-free period were not significantly related to the serum HA level, the serum HA level was significantly associated with overall survival after hepatectomy (p 0.05). Cox’s multivariate analysis did not show a significant relationship between HA level and survival. Serum HA reflects progression and survival in HCC patients.