Okayama University Medical SchoolActa Medica Okayama0386-300X6942015Local Recurrence and Complications after Percutaneous Radiofrequency Ablation of Hepatocellular Carcinoma : A Retrospective Cohort Study Focused on Tumor Location219226ENJunichiToshimoriKazuhiroNousoShinichiroNakamuraNozomuWadaYukiMorimotoYasutoTakeuchiTetsuyaYasunakaKenjiKuwakiHidekiOhnishiFusaoIkedaHidenoriShirahaAkinobuTakakiKazuhideYamamotoOriginal Article10.18926/AMO/53558We conducted a retrospective cohort study to investigate the predisposing factors for local recurrence and complications after percutaneous radiofrequency ablation (RFA) of hepatocellular carcinoma (HCC). HCC patients (n＝397) consecutively treated with RFA (256 males, 141 females, median age 69 years) were enrolled. In these patients, 1,455 nodules (median size 17mm) were ablated. Predisposing factors for overall recurrence and local recurrence in the context of tumor location and complications were examined. Local recurrence was observed for 113 of the 1,455 nodules. The 1-, 3- and 5-year local recurrence rates were 2.2ｵ, 7.4ｵ and 9.5ｵ, respectively. A multivariate Cox proportional hazard analysis revealed that large tumor size (＞2cm), tumor location (adjacent to the major portal branch or hepatic vein), and small ablated margin (＜3mm) were independent predisposing factors for local recurrence after RFA (HR＝1.70-2.81). Tumor location (adjacent to the major portal branch, hepatic vein, or diaphragm) was also revealed as a risk factor for liver damage due to RFA. HCC adjacent to the major portal vein or hepatic vein was associated with a higher risk for local recurrence and for complications;therefore, special precautions are necessary when applying RFA to HCC near vessels even when the tumors are located at an easy-to-puncture site.No potential conflict of interest relevant to this article was reported.Acta Medica Okayama0944-11744832013The impact of patatin-like phospholipase domain-containing protein 3 polymorphism on hepatocellular carcinoma prognosis405412ENYasutoTakeuchiFusaoIkedaYukiMoritouHiroakiHagiharaTetsuyaYasunakaKenjiKuwakiYasuhiroMiyakeHidekiOhnishiShinichiroNakamuraHidenoriShirahaAkinobuTakakiYoshiakiIwasakiKazuhiroNousoKazuhideYamamotoThe single nucleotide polymorphism (SNP) rs738409 in patatin-like phospholipase domain-containing protein 3 (PNPLA3) is associated with hepatic fat accumulation and disease progression in patients with non-alcoholic fatty liver disease and alcoholic liver disease (ALD). This study was conducted to determine whether PNPLA3 rs738409 SNPs affect development and prognosis of hepatocellular carcinoma (HCC) in patients with various liver diseases.
We enrolled 638 consecutive Japanese patients newly diagnosed with HCC between 2001 and 2010: 72 patients with hepatitis B virus (HBV), 462 with hepatitis C virus (HCV), and 104 with non-B non-C (NBNC).
NBNC patients exhibited large tumors of advanced TNM stages at HCC diagnosis, and had significantly poorer prognosis than HBV or HCV patients (P < 0.001 and < 0.001, respectively; log-rank test). The G/G genotype of PNPLA3 rs738409 SNP had significantly higher distribution in NBNC patients (P < 0.001) and was significantly associated with higher body mass index (BMI) and an increased aspartate aminotransferase to platelet ratio index. No significant differences were observed in survival with differences in PNPLA3 SNP genotypes among the patients, although ALD patients with the G/G genotype of PNPLA3 SNP and low BMI had significantly poorer survival than those with high BMI (P = 0.028).
The G/G genotype of PNPLA3 rs738409 SNP was more frequently distributed, and associated with BMI and fibrosis among NBNC-HCC patients but not among HBV or HCV patients. These genotypes might affect HCC prognosis in ALD patients, but not in HBV, HCV, or NAFLD patients.No potential conflict of interest relevant to this article was reported.Acta Medica Okayama0944-11744742012Risk factors for recurrence after transarterial chemoembolization for early-stage hepatocellular carcinoma421426ENHideakiKinugasaKazuhiroNousoYasutoTakeuchiTetsuyaYasunakaHidekiOnishiShin-ichiroNakamuraHidenoriShirahaKenjiKuwakiHiroakiHagiharaFusaoIkedaYasuhiroMiyakeAkinobuTakakiKazuhideYamamotoRadiofrequency ablation (RFA) is a standard therapy for the treatment of hepatocellular carcinoma (HCC) with 3 or fewer tumors of up to 3 cm (early-stage HCC); when RFA is unsuccessful or unfeasible, transcatheter arterial chemoembolization (TACE) has often been performed. However, little information about the outcome of TACE for early-stage HCC has been reported and it is hard to decide whether to perform additional treatment following TACE in these difficult conditions. The aim of this study was to determine the risk factors for local or intrahepatic distant recurrence after TACE in early-stage HCC.
Among 1,560 newly diagnosed HCC patients who were admitted to Okayama University Hospital, 43 patients with early-stage HCC who received only TACE in at least one nodule were enrolled in this study. We analyzed the risk factors for local and distant recurrence by the Cox proportional hazard model.
The local recurrence rates and intrahepatic distant recurrence rates at 3 months, 6 months, and 1 year were 18.6, 33.4, and 61.8%, and 2.8, 2.8, and 10.2%, respectively. Among 12 parameters examined as possible risk factors for recurrence, heterogeneous Lipiodol uptake (risk ratio 3.38; 95% confidence interval 1.14-10.60) and high serum des-gamma-carboxy prothrombin (DCP) (2.58; 1.03-7.14) were significantly correlated with local recurrence, and the presence of multiple tumors (10.64; 1.76-93.75) was significantly correlated with intrahepatic distant recurrence.
Heterogeneous Lipiodol uptake, high serum DCP, and multiple tumors are risk factors for recurrence in patients with early-stage HCC who have undergone palliative TACE.No potential conflict of interest relevant to this article was reported.Acta Medica Okayama0815-931927102012Des-gamma-carboxyl prothrombin is associated with tumor angiogenesis in hepatocellular carcinoma16021608ENMinoruMatsubaraHidenoriShirahaJyunroKataokaMasayaIwamuroShigeruHoriguchiShin-ichiNishinaNobuyukiTakaokaMasayukiUemuraAkinobuTakakiShinichiroNakamuraYoshiyukiKobayashiKazuhiroNousoKazuhideYamamotoBackground and Aim: Hepatocellular carcinoma (HCC) is a hypervascular tumor, and angiogenesis plays an important role in its development. Previously, we demonstrated that des-gamma-carboxyl prothrombin (DCP) promotes both cell proliferation and migration of human umbilical vein endothelial cells (HUVECs) by inducing the autophosphorylation of kinase insert domain receptor (KDR). In the present study, DCP-associated tumor angiogenesis was assessed by comparing hypovascular and common hypervascular HCC. Methods: The solitary HCCs of 827 patients were classified into two groups according to the tumor density at the arterial phase of a dynamic computed tomography scan; the initial clinical data of patients with the hyper- and hypovascular types were compared. The HCC tissues from 95 tumors were analyzed by immunohistochemical staining for DCP and phosphorylated KDR, and intratumoral microvessel density (MVD) was analyzed to evaluate microvessel angiogenesis. Results: The serum DCP levels (320 +/- 3532 mAU/mL) and tumor size (18.4 +/- 9.0 mm) of patients with hypervascular HCC were significantly greater than those with hypovascular HCC (38.7 +/- 80 mAU/mL and 14.6 +/- 5.2 mm, P < 0.001). Immunohistochemical analysis revealed that the expressions of DCP and phospho-KDR were significantly greater in hypervascular HCC (71.4% and 31.0%, respectively) than in hypovascular HCC (7.6% and 5.7%, respectively). Intratumoral MVD was significantly correlated with DCP (r = 0.48, P < 0.0001). Conclusions: des-gamma-carboxyl prothrombin production is associated with tumor angiogenesis in HCC.No potential conflict of interest relevant to this article was reported.Wiley-BlackwellActa Medica Okayama1386-634643102013Serum oxidative-anti-oxidative stress balance is dysregulated in patients with hepatitis C virus-related hepatocellular carcinoma10781092ENMamoruNishimuraAkinobuTakakiNaofumiTamakiTakayukiMaruyamaHidekiOnishiSayoKobayashiKazuhiroNousoTetsuyaYasunakaKazukoKoikeHiroakiHagiharaKenjiKuwakiShinichiroNakamuraFusaoIkedaYoshiakiIwasakiTakaakiTomofujiManabuMoritaKazuhideYamamotoAim
Oxidative stress is associated with progression of chronic liver disease (CLD). This association is best established in chronic hepatitis C. However, the anti-oxidative state is not well characterized. The objective of the present study was to investigate the balance of oxidative and anti-oxidative stress in CLD patients.
Methods
We recruited a study population of 208 patients, including healthy volunteers (HV; n = 15), patients with hepatitis B virus (HBV)-related CLD without or with hepatocellular carcinoma (HBV-non-HCC, n = 25, and HBV-HCC, n = 50, respectively), and patients with hepatitis C virus (HCV)-related CLD without or with HCC (HCV-non-HCC, n = 49, and HCV-HCC, n = 69, respectively). Serum levels of reactive oxygen metabolites (ROM) and anti-oxidative markers (OXY-adsorbent test; OXY) were determined, and the balance of these values was used as the oxidative index. Correlations among ROM, OXY, oxidative index and clinical characteristics were investigated.
Results
Patients with CLD exhibited elevated ROM and oxidative index compared to HV. Among patients with CLD, HCV positive status correlated with increased ROM. In CLD, HCV-HCC patients exhibited the highest ROM levels. Among HCV-related CLD patients, lower OXY correlated with HCC positive status, but was recovered by eradication of HCC. In HCV-HCC, lower OXY correlated with high PT-INR.
Conclusion
HCV positive CLD patients displayed higher oxidative stress and HCV-HCC patients displayed lower anti-oxidative state. Anti-oxidative state depression was associated with liver reservoir-related data in HCV-HCC and could be reversed with HCC eradication.No potential conflict of interest relevant to this article was reported.Acta Medica Okayama0020-7136131112012Runt-related transcription factor 3 reverses epithelial-mesenchymal transition in hepatocellular carcinoma25372546ENShigetomiTanakaHidenoriShirahaYutakaNakanishiShin-IchiNishinaMinoruMatsubaraShigeruHoriguchiNobuyukiTakaokaMasayaIwamuroJunroKataokaKenjiKuwakiHiroakiHagiharaJunichiToshimoriHidekiOhnishiAkinobuTakakiShinichiroNakamuraKazuhiroNousoTakahitoYagiKazuhideYamamotoLoss or decreased expression of runt-related transcription factor 3 (RUNX3), a tumor suppressor gene involved in gastric and other cancers, has been frequently observed in hepatocellular carcinoma (HCC). The objective of this study was to identify the regulatory mechanism of the epithelialmesenchymal transition (EMT) by RUNX3 in HCC. Human HCC cell lines, Hep3B, Huh7, HLF and SK-Hep1, were divided into low- and high-EMT lines, based on their expression of TWIST1 and SNAI2, and were used in this in vitro study. Ectopic RUNX3 expression had an anti-EMT effect in low-EMT HCC cell lines characterized by increased E-cadherin expression and decreased N-cadherin and vimentin expression. RUNX3 expression has previously been reported to reduce jagged-1 (JAG1) expression; therefore, JAG1 ligand peptide was used to reinduce EMT in RUNX3-expressing low-EMT HCC cells. Immunohistochemical analyses were performed for RUNX3, E-cadherin, N-cadherin and TWIST1 in 33 human HCC tissues, also divided into low- and high-EMT HCC, based on TWIST1 expression. E-cadherin expression was correlated positively and N-cadherin expression was correlated negatively with RUNX3 expression in low-EMT HCC tissues. Correlations between EMT markers and RUNX3 mRNA expression were analyzed using Oncomine datasets. Similarly, mRNA expression of E-cadherin was also significantly correlated with that of RUNX3 in low-EMT HCC, while mRNA expression of JAG1 was negatively correlated with that of RUNX3. These results suggest a novel mechanism by which loss or decreased expression of RUNX3 induces EMT via induction of JAG1 expression in low-EMT HCC.No potential conflict of interest relevant to this article was reported.Acta Medica Okayama1471-2407112011Loss of runt-related transcription factor 3 expression leads hepatocellular carcinoma cells to escape apoptosisENYutakaNakanishiHidenoriShirahaShin-ichiNishinaShigetomiTanakaMinoruMatsubaraShigeruHoriguchiMasayaIwamuroNobuyukiTakaokaMasayukiUemuraKenjiKuwakiHiroakiHagiharaJunichiToshimoriHidekiOhnishiAkinobuTakakiShinichiroNakamuraYoshiyukiKobayashiKazuhiroNousoTakahitoYagiKazuhideYamamotoBackground: Runt-related transcription factor 3 (RUNX3) is known as a tumor suppressor gene for gastric cancer and other cancers, this gene may be involved in the development of hepatocellular carcinoma (HCC).
Methods: RUNX3 expression was analyzed by immunoblot and immunohistochemistry in HCC cells and tissues, respectively. Hep3B cells, lacking endogenous RUNX3, were introduced with RUNX3 constructs. Cell proliferation was measured using the MTT assay and apoptosis was evaluated using DAPI staining. Apoptosis signaling was assessed by immunoblot analysis.
Results: RUNX3 protein expression was frequently inactivated in the HCC cell lines (91%) and tissues (90%). RUNX3 expression inhibited 90 +/- 8% of cell growth at 72 h in serum starved Hep3B cells. Forty-eight hour serum starvation-induced apoptosis and the percentage of apoptotic cells reached 31 +/- 4% and 4 +/- 1% in RUNX3-expressing Hep3B and control cells, respectively. Apoptotic activity was increased by Bim expression and caspase-3 and caspase-9 activation.
Conclusion: RUNX3 expression enhanced serum starvation-induced apoptosis in HCC cell lines. RUNX3 is deleted or weakly expressed in HCC, which leads to tumorigenesis by escaping apoptosis.No potential conflict of interest relevant to this article was reported.Acta Medica Okayama1471-240792009Twist expression promotes migration and invasion in hepatocellular carcinomaENNoriyukiMatsuoHidenoriShirahaTatsuyaFujikawaNobuyukiTakaokaNaokiUedaShigetomiTanakaShinichiNishinaYutakaNakanishiMasayukiUemuraAkinobuTakakiShinichiroNakamuraYoshiyukiKobayashiKazuhiroNousoTakahitoYagiKazuhideYamamotoBackground: Twist, a transcription factor of the basic helix-loop-helix class, is reported to regulate cancer metastasis. It is known to induce epithelial-mesenchymal transition (EMT). In this study, we evaluated the expression of twist and its effect on cell migration in hepatocellular carcinoma (HCC).
Methods: We examined twist expression using immunohistochemistry in 20 tissue samples of hepatocellular carcinoma, and assessed twist expression in HCC cell lines by RT-PCR and Western blot analysis. Ectopic twist expression was created by introducing a twist construct in the twist-negative HCC cell lines. Endogenous twist expression was blocked by twist siRNA in the twist-positive HCC cell lines. We studied EMT related markers, E-cadherin, Vimentin, and N-cadherin by Western blot analysis. Cell proliferation was measured by MTT assay, and cell migration was measured by in vitro wound healing assay. We used immunofluorescent vinculin staining to visualize focal adhesion.
Results: We detected strong and intermediate twist expression in 7 of 20 tumor samples, and no significant twist expression was found in the tumor-free resection margins. In addition, we detected twist expression in HLE, HLF, and SK-Hep1 cells, but not in PLC/RPF/5, HepG2, and Huh7 cells. Ectopic twist-expressing cells demonstrated enhanced cell motility, but twist expression did not affect cell proliferation. Twist expression induced epithelial-mesenchymal transition together with related morphologic changes. Focal adhesion contact was reduced significantly in ectopic twist-expressing cells. Twist-siRNA-treated HLE, HLF, and SK-Hep1 cells demonstrated a reduction in cell migration by 50, 40 and 18%, respectively.
Conclusion: Twist induces migratory effect on hepatocellular carcinoma by causing epithelial-mesenchymal transition.No potential conflict of interest relevant to this article was reported.Okayama University Medical SchoolActa Medica Okayama0386-300X6512011Prognostic Model for Hepatocellular Carcinoma with Time-Dependent Factors1119ENKenjiKuwakiKazuhiroNousoYoshiyukiKobayashiShinichiroNakamuraYoichi M.ItoShoutaIwadouHiroakiHagiharaTetsuyaYasunakaJunichiToshimoriHirokazuMiyatakeKenjiMiyoshiHidekiOnishiYasuhiroMiyakeBonShojiAkinobuTakakiHidenoriShirahaYoshiakiIwasakiHaruhikoKobashiKazuhideYamamotoOriginal Article10.18926/AMO/43825The purpose of this study was to build a prognostic model of hepatocellular carcinoma (HCC) using time-dependent covariates to re-evaluate the prognosis at any stage of the disease. The subjects were consecutive HCC patients who were treated at our institute between 1995 and 2007. We constructed time-fixed and time-dependent prognostic models with a training group (n＝336) and compared the prognostic abilities between conventional Cancer of the Liver Italian Program (CLIP) scores, Japan Integrated Staging (JIS) scores, an Okuda classification, and our prognostic models in the testing group (n＝227) with the c-index. The time-dependent prognostic model consisted of main tumor size, tumor number, portal vein invasion, distant metastasis, alpha-fetoprotein, des-gamma-carboxy prothrombin (DCP), bilirubin, and albumin and the weighted scores were set for each factor depending on the hazard ratio for the prognosis. The prognostic index was determined by summing the scores. The c-index values for the CLIP scores, JIS scores, Okuda classification, and our time-dependent model were 0.741, 0.727, 0.609, and 0.870, respectively. These results indicate that our time-dependent model can estimate the prognosis of HCC more precisely than traditional time-fixed models and can be used to re-predict the prognosis of HCC.No potential conflict of interest relevant to this article was reported.岡山医学会Acta Medica Okayama0030-155812112009肝癌診療ガイドライン4145ENYoshiyukiKobayashiShinichiroNakamuraHidekiOhnishiJunichiToshimoriKenjiKuwakiHiroakiHagiharaYasuhiroMiyakeHidenoriShirahaKazuhiroNousoTakahitoYagiNoriakiTanakaKazuhideYamamotoNo potential conflict of interest relevant to this article was reported.岡山医学会Acta Medica Okayama0030-155811332001肝細胞癌の超音波ガイド下ラジオ波焼灼療法による治療289294ENKazuhiroNousoYoshiyukiKobayashiShinichiroNakamuraEiichiroYumotoTakahiroSuzukiTomomiHakodaYoshitakaTakumaHironoriTanakaEijiMatsumotoHideakiTaniguchiTatsuyaFujikawaMayumiSuzukiAkioMoriyaKousakuSakaguchiTakaoTsujiHapatocellular carcinoma (HCC) is one of the most prevalent malignancies in Japan. Percutaneous ethanol injection therapy (PEIT) of microwave coagulation therapy (MCT) have been used to treat small HCC. These two methods are effective methods to treat HCC; however, they have weak points. A period of admission is long to treat patients with PEIT because it requires several sessions to kill HCC completely. The risk of complications of MCT is high because the needle is thick, although it can coagulate HCC completely in one session. Radio frequecy ablation (RFA) is a new method to treat HCC. RFA compensates these weak points and is thought to become a main modality to treat HCC. In this paper, we demonstrated two patients with HCC who were treated with RFA. The effect of the treatment was examined with helical computed tomography and contrast harmonic power doppler, and the results were satisfactory. We also mentioned the merits and demerits of RAF in Discussion.No potential conflict of interest relevant to this article was reported.Acta Medica Okayama2006Expression and immunogenicity of NY-ESO-1 in hepatocellular carcinomaENShin-ichiroNakamuraNo potential conflict of interest relevant to this article was reported.