Sustained low alanine aminotransferase levels can predict the survival for 10 years without hepatocellular carcinoma development in patients with hepatitis C virus-associated liver cirrhosis of child stage A

An analysis was performed of the patients with hepatitis C virus-associated liver cirrhosis (HCV-LC) who never developed hepatocellular carcinoma (HCC) for 10 years after the histological diagnosis of LC. Seventy-four consecutive HCV-LC patients of Child stage A were observed for >10 years prospectively for the development of HCC with frequent ultrasonography and magnetic resonance imaging or computed tomography. Of the 63 patients who fulfilled the study, 48 patients were treated and 15 were nontreated because of their stable state. They were subdivided into three groups according to their serum alanine aminotransferase (ALT) levels: the high ALT group comprised of 23 patients whose annual average serum ALT level was persistently high (>/=80 IU); the low ALT group comprised of 28 patients whose annual average serum ALT level was persistently low (<80 IU), and the unclassified ALT group comprised of 12 patients. In the low ALT group, as high as 71.4% of patients had never developed HCC for 10 years, in contrast to only 17.4% in the high ALT group (p < 0.001). In the 30 patients who never developed HCC for 10 years, 20 patients belonged to the low ALT group, in contrast to only 4 belonging to the high ALT group. Sustained low ALT levels were important to survive for 10 years without developing HCC in the HCV-LC patients of Child stage A.     

Detection of TT virus (TTV) in non-B, non-C hepatitis patients with hepatocellular carcinoma, and clinical features of these patients]

A novel human DNA virus, TTvirus (TTV), was identified from a patient with posttransfusion hepatitis of unknown etiology. It is thought to be a new hepatitis virus, but the clinical significance of this virus is uncertain. We investigated the frequency of TTV viremia by PCR in 39 non-B, non-C hepatitis (NBNC) patients with hepatocellular carcinoma (HCC), and clinical features of these patients. TTV viremia was detected in 20 (51.3%) of 39 NBNC hepatitis patients with HCC. Liver cirrhosis (LC) were found in 11 (55%) of 20 TTV-positive patients and 16 (84%) of 19 TTV-negative patients (p < 0.05). The levels of AST, LDH, LAP, gamma GTP in TTV-positive patients were significantly higher than those in TTV-negative patients (p < 0.05). (AST: 58 +/- 26 vs 42 +/- 23 IU/l, LDH: 468 +/- 127 vs 366 +/- 123 IU/l, LAP: 339 +/- 242 vs 206 +/- 80 IU/l, gamma GTP: 207 +/- 207 vs 105 +/- 107 IU/l) These results suggest clinical differences between TTV-positive and TTV-negative patients in NBNC hepatitis patients with HCC.     

肝癌肝移植后亚砷酸化疗的应用

背景：移植后肿瘤复发是影响肝癌肝移植疗效的主要因素,如何防止肝癌肝移植后肿瘤复发是目前肝移植研究的热点问题之一。亚砷酸全身化疗被认为对中晚期肝癌具有一定作用,但在肝移植后的应用还未见报道。目的：观察超出米兰标准的肝癌患者肝移植后应用亚砷酸全身化疗的对肿瘤复发的干预效果。方法：对23例超出米兰标准的肝癌患者肝移植后采用亚砷酸行预防性化疗：静脉滴注10mg/d,连续使用7d后间隔7d,重复4次为1个疗程,患者接受1~4个疗程。观察以上使用亚砷酸化疗患者的生存、肿瘤复发情况,以及化疗不良反应,并与同期16例未使用化疗的肝癌肝移植患者相比较。结果与结论：经过3~32个月随访,共30例患者出现肝癌复发,化疗组16例,非化疗组14例,复发部位最常见于肺部、移植肝及骨骼。化疗组与非化疗组肿瘤复发率差异无显著性意义,但化疗组复发时间明显延迟（P=0.026）;两组6个月、1年生存率差异无显著性意义,化疗组2年生存率显著高于非化疗组（P=0.037）;两组6个月无瘤生存率差异无显著性意义,1年、2年无瘤生存率化疗组显著高于明显非化疗组（P=0.030,0.023）。亚砷酸使用过程中未发现严重不良反应。提示肝癌肝移植患者静脉使用亚砷酸化疗可以延迟肿瘤复发,提高生存率。     

Methylated cyclin D2 gene circulating in the blood as a prognosis predictor of hepatocellular carcinoma

BackgroundPrognosis of hepatocellular carcinoma (HCC) remains poor because of high recurrence rate. We examined preoperatively the methylated CCND2 gene levels present in the serum following release from HCC cells as a prognosis predictor in patients undergoing curative hepatectomy.MethodsQuantitative real-time RT-PCR and quantitative methylation-specific PCR were used to measure methylated CCND2 gene and its mRNA levels.ResultsThe CCND2 mRNA levels were down-regulated in HCC with early intrahepatic recurrence (IHR) within 1 year of curative hepatectomy. We also identified that this down-regulation was due to promoter hypermethylation. In 70 HCC patients who underwent curative hepatectomy, 39 patients sero-positive for the methylated CCND2 gene (> 70 pg/ml serum) exhibited a significantly shorter disease-free survival (DFS) period (P = 0.02) than the 31 patients who were sero-negative for the methylated CCND2 gene. None of the sero-negative patients demonstrated early IHR, and this method of serum testing did not produce any false-negative predictions for early IHR. Multivariate analysis showed that the serum level of methylated CCND2 was an independent risk factor for DFS (hazard ratio of 1.866, 95% CI: 1.106–3.149).ConclusionMethylated CCND2 gene in the serum serves as a prognosis predictor of HCC after curative hepatectomy.     

Hepatitis B virus-related hepatocellular carcinogenesis and its prevention

To elucidate the influence of serum hepatitis B virus (HBV) load on hepatocellular carcinogenesis in cirrhotic patients, HBV-DNA was sequentially measured. In a nested, case-control study using 96 patients without antiviral therapy, high HBV-DNA (> or =10(3.7) copies/ml) in the last 3 years was significantly associated with carcinogenesis (a patient group without hepatocellular carcinoma (HCC) development; 0/48 vs. a patient group with eventual HCC development; 22/48, p < 0.0001). No patient with a continuously low HBV-DNA for the last 3 years developed HCC. Persistence of high HBV-DNA concentration suggested an increased risk of carcinogenesis. In a retrospective cohort study using 57 patients with interferon therapy, HCC developed in 2 (8.0%) of the 25 patients with HBV-DNA loss, while carcinogenesis was found in 11 (34.4%) of 32 patients without HBV-DNA loss (Fisher's exact test, p = 0.026). A significant decrease or loss of serum HBV-DNA stops HCC development, and its sequential analysis could be very useful both in the prediction and early detection of small HCC.     

Possible contribution of serum activin A and IGF-1 in the development of hepatocellular carcinoma in Egyptian patients suffering from combined hepatitis C virus infection and hepatic schistosomiasis

OBJECTIVES: The present study evaluated the role of activin A, insulin-like growth factor 1 (IGF-1) and insulin-like growth factor binding protein 3 (IGFBP-3) in Egyptian patients suffering from combined hepatitis C virus (HCV) infection and hepatic schistosomiasis. DESIGN AND METHODS: Four groups were included in the present study. Group I: 30 healthy subjects were included as controls; Group II (HCV): 30 patients with chronic liver disease due to HCV infection without evidence of schistosomiasis; Group III (SHF + HCV): 30 patients with combined disease, chronic schistosomal hepatic fibrosis (SHF) and chronic hepatitis C infection; Group IV (HCC): 30 patients with hepatocellular carcinoma associated with chronic hepatitis C virus and schistosomal infection. RESULTS: Patients with HCV, HCV + SHF and those with HCC had a significantly higher serum activin A compared with the control group (P < 0.001). Serum activin A level (mean +/- SD) was 5.7 +/- 2.76, 10.59 +/- 3.59, 15.39 +/- 4.61 and 19.93 +/- 5.43 ng/mL in controls, HCV patients, HCV + SHF patients and HCC patients, respectively. Serum IGF-1 was significantly lower in HCV patients, HCV + SHF patients and HCC patients compared to the control group (P < 0.001). Serum IGF-1 was 121.7 +/- 73.4, 76.7 +/- 23.5, 35.7 +/- 17.6 and 39.9 +/- 25.9 ng/mL in controls, HCV patients, HCV + SHF patients and HCC patients, respectively. Similarly, serum IGFBP-3 was significantly lower in HCV patients, HCV + SHF patients and HCC patients compared to the control group (P < 0.001). Furthermore, serum insulin-like growth factor binding protein 3 (IGFBP-3) was significantly lower in patients with HCC compared to patients with HCV or those with HCV + SHF (P < 0.01 and P = 0.024, respectively). The median (range) of serum IGFBP-3 was 4452 (352.2-8965), 3457 (1114-6000), 2114 (867-5901) and 1202 (576-3994) ng/mL in controls, HCV patients, HCV + SHF patients and HCC patients, respectively. Serum activin A correlated positively with Child-Pugh scoring in patients with HCV, HCV + SHF and those with HCC. The correlation coefficient was significant, at 0.001, in total cases. CONCLUSIONS: We conclude that patients with HCV, HCV + SHF and those with HCC have a significantly higher serum activin A when compared with controls. Serum activin A level was significantly higher in patients with HCV + SHF compared to those with HCV alone (P < 0.01) with a significant positive correlation between the serum activin A level and Child-Pugh scoring in patients with HCV, HCV + SHF and those with HCC. Furthermore, serum IGF-1 and IGFBP-3 levels were significantly reduced in patients with HCV, HCV + SHF and those with HCC compared to the control group. We suggest that this pattern (high activin A and low IGF-1 and its binding protein 3) may play a role in development of HCC in Egyptian patients suffering from combined hepatitis C virus infection and hepatic schistosomiasis.     

Diabetes worsens the surgical outcomes in cirrhotic patients with hepatocellular carcinoma

Diabetes is a known risk factor for developing hepatocellular carcinoma (HCC). Reported rates of diabetes are higher in chronic hepatitis, cirrhosis and HCC patients. However, its effects on postoperative recurrence and survival with HCC are controversial. This study offers a retrospective analysis of the impacts of diabetes on postoperative recurrence and survival in patients with cirrhosis and HCC. A total of 389 cirrhotic patients who underwent curative resection for primary HCC at our institution between January 2000 and December 2008 were enrolled. Of them, 272 (70%) patients were classified into a non-diabetes group and 117 (30%) patients into the diabetes group. The diabetes group was divided into an oral hypoglycemic agent (OHA) control group (n = 100) and an insulin control group (n = 17). The result indicates that the diabetes group had a higher postoperative recurrence rate and poorer long-term survival rate (p = 0.001 vs. 0.01). There was no significant difference in recurrence-free survival rate between the OHA control group and the insulin control group (p = 0.17). The insulin control group had a poorer long-term surgical outcome than the OHA control group (p = 0.035). In conclusion, our results suggest that diabetes is an independent risk factor for postoperative recurrence and surgical survival of cirrhotic HCC patients. Cirrhotic HCC patients with diabetes who received hepatic resection should be closely followed-up for postoperative recurrence and long-term outcome. Moreover, an effective peri-operative sugar control planning in HCC patients with diabetes should be established.     

Metastasis-related miR-185 is a potential prognostic biomarker for hepatocellular carcinoma in early stage

We previously reported that miR-185 is associated with hepatocellular carcinoma (HCC) venous metastasis analysed by miRNA-array profile. The aim of this study is to further investigate the clinicopathological significance and prognostic value of miR-185 in early stage HCC. We classified 95 patients with early stage HCC into treated recurrence group (TR) and none treated recurrence group (NTR), and detected the miR-185 expression levels in TR and NTR groups. We found that low miR-185 expression correlated with more tumor recurrence (37/46), while high miR-185 level led to lower recurrence rate (17/49) (P < 0.05). There was no direct relationship between miR-185 and clinicopathological features, including age, gender, ALT, AFP, liver cirrhosis, tumor size, tumor encapsulation, tumor differentiation (P > 0.05). Kaplan–Meier analysis showed that low miR-185 group had a remarkable lower survival rate and shorter time to recurrence than high miR-185 group (P < 0.05). Univariate and multivariate analysis, using Cox's proportional hazards model, also indicated that low miR-185 expression was a sensitive prognostic factor for survival and recurrence in early stage HCC (P < 0.05). We upregulated or downregulated miR-185 expression by transfected miR-185 mimics or inhibitor into HCC cell lines, and observed the influence of miR-185 on HCC cells in vitro. Our results manifested that miR-185 could suppress the tumor cell growth and invasive ability (P < 0.05). Therefore, miR-185 might be an effective and sensitive biomarker of HCC in early stage, and the upregulation of miR-185 might be considered to be a potentially important molecular treatment strategy for patients with HCC.     

利用芯片技术筛选肝癌早期复发密切相关的microRNA

目的:应用miRNA芯片技术筛选肝癌早期复发密切相关的miRNA,为进一步深入探讨miRNA在介导肝癌高侵袭性及术后早期复发中的作用打下基础。方法:从本院肝癌标本库中选取10例肝癌患者进入本研究,其中早期复发组(术后第1年出现肝内复发病灶)和非早期复发组(术后2年以上未出现肝癌复发或转移)各5例,抽提总RNA并分离出小分子RNA进行Cy3荧光标记,将标记的小分子RNA与miRNA芯片进行杂交反应,分析差异表达的miRNA。结果:相对于非早期复发组,早期复发组肿瘤标本中有miR-602、miR-451、miR-144和miR-486-5p等4个miRNAs表达显著上调(上调倍数>2.0);有miR-551b、miR-96和miR-502-3p等3个miRNAs表达显著下调(下调倍数<0.5)。结论:筛选得到肝癌早期复发的miRNA差异表达谱,其可能与肝癌的早期复发发生发展有关。     

Risk factors for long-term prognosis of hepatocellular carcinoma patients after anatomic hepatectomy

BACKGROUND The risk factors for patients with major postoperative complications immediately after liver resection have been identified;however,the intermediate and longterm prognoses for these patients have yet to be determined.AIM To evaluate the factors responsible for the long-term recurrence-free survival rate in patients with hepatocellular carcinoma(HCC)following anatomic hepatectomy.METHODS We performed a retrospective analysis of 74 patients with HCC who underwent precise anatomic hepatectomy at our institution from January 2013 to December 2015.The observational endpoints for this study were the tumor recurrence or death of the HCC patients.The overall follow-up duration was three years.The recurrence-free survival curves were plotted by the Kaplan-Meier method and were analyzed by the log-rank test.The value of each variable for predicting prognosis was assessed via multivariate Cox proportional hazards regression analysis.RESULTS The 1-year and 3-year recurrence-free survival rates of HCC patients were 68.92%and 55.41%,respectively,following anatomic liver resection.The results showed that the 3-year recurrence-free survival rate in HCC patients was closely related to preoperative cirrhosis,jaundice level,tumor stage,maximal tumor diameter,complications of diabetes mellitus,frequency of intraoperative hypotensive episodes,estimated blood loss(EBL),blood transfusion,fluid infusion,and postoperative infection(P<0.1).Based on multivariate analysis,preoperative cirrhosis,tumor stage,intraoperative hypotension,and EBL were identified to be predictors of 3-year recurrence-free survival in HCC patients undergoing anatomic hepatectomy(P<0.05).CONCLUSION Tumor stage and preoperative cirrhosis adversely affect the recurrence-free survival rate in HCC patients following anatomic hepatectomy.The long-term recurrence-free survival rate of patients with HCC is closely related to intraoperative hypotension and EBL.     

影响肝细胞癌根治性切除术后预后因素的Cox模型分析

目的探讨影响肝细胞癌根治性切除术后预后的相关因素，为肝细胞癌的术后综合治疗及判断预后提供依据。方法回顾性分析194例肝细胞癌的临床病理资料，对潜在的可能影响术后预后的临床病理因素进行单因素分析，将单因素分析后有统计学意义的因素带人Cox比例风险回归模型进行多因素分析。结果全组患者的术后1、3、5年的累积生存率分别为81．4％、50．6％、33．9％。单因素分析结果显示：术前AST、ALP、GGT、肿瘤最大直径、肿瘤病灶数目、手术持续时间、手术中总失血量、手术中输血情况、术后2年内复发是有统计学意义的因素。Cox模型多因素分析结果示术后2年内复发、术前AST浓度是有统计学意义的因素。结论HCC根治性切除术后的预后取决于多种因素的共同作用，术后2年内复发、术前AST浓度是影响术后预后的最重要因素。     

Recurrent hepatocellular carcinoma: usefulness of ultrasonography compared with computed tomography and AFP assay.

Follow-up studies using monitoring with alpha1-feto protein (AFP), ultrasonography (US), and computed tomography (CT) were carried out in 75 patients who had prior partial hepatectomy for hepatocellular carcinoma (HCC). Recurrence in the remaining liver was confirmed in 31 patients (41.3%) during the 4 month to 3 years 7 month period. Ultrasonography detected recurrence in 29 cases (sensitivity: 93.5%), CT in 26 (83.9%), and AFP assay in 12 (38.7%). In 4 patients, ultrasonography detected four recurrent nodules that CT missed. In 1 patient, two subphrenic nodules were detected with CT but not with ultrasonography. The specificity of US, CT, and AFP assay was 90.9%, 95.5%, and 93.2% respectively. Frequent follow-up study with ultrasonography in combination with CT and AFP assay should be recommended for the early detection of recurrent HCC. Ultrasonography is mandatory for the follow-up of patients with a prior hepatectomy for HCC.     

磷脂酰肌醇蛋白多糖-3在肝细胞肝癌的表达及其与术后复发的相关性

目的探讨磷脂酰肌醇蛋白多糖-3（GPC-3）mRNA在肝细胞癌（HCC）癌组织及癌旁组织的表达及其与术后复发的相关性。方法用逆转录聚合酶链反应测定60例HCC癌组织及癌旁组织中GPC-3 mRNA的表达。PCR产物经凝胶电泳,比较HCC组织及癌旁组织中GPC-3与-βactin条带的灰度值之比,半定量分析GPC-3 mRNA的表达水平,分析术后3年不易复发HCC患者的癌组织GPC-3 mRNA的表达水平。结果 GPC-3 mRNA在HCC癌组织中高度表达,阳性率为68.3%,癌旁组织中低表达或未见表达,术后3年内未复发的HCC患者癌组织中GPC-3 mRNA呈现低表达水平,GPC-3 mRNA表达与肿瘤大小、肿瘤数目、有无包膜、门静脉癌栓、HBsAg及AFP水平无明显相关。结论 GPC-3 mRNA在HCC癌组织中的表达明显高于癌旁组织,可能是HCC的一种新的潜在的肿瘤特异标志物;癌组织中GPC-3mRNA低表达的HCC患者不易复发,可能是判断HCC进展和评价预后的一个潜在重要指标。     

Adoptive immunotherapy for postoperative hepatocellular carcinoma: a systematic review

The high risk of recurrence in post-operative hepatocellular carcinoma (HCC) highlights the need for an effective adjuvant treatment. A systematic review of randomised controlled trials (RCTs) was performed to evaluate the clinical efficacy of adjuvant adoptive immunotherapy (AIT) for post-operative HCC patients. Electronic (MEDLINE, EMBASE and Cochrane Library databases) and manual searches were conducted throughout May 2011 to identify RCTs evaluating postoperative AIT for patients with HCC. Methodological quality was assessed in accordance with the QUOROM statement. Four RCTs totalling 423 patients met the eligibility criteria. All RCTs reported significantly improved disease-free survival rate or reduced recurrence rate after treating with adjuvant AIT (p < 0.05). The overall survival rates of AIT group are slightly higher than those of the control group in one study. Moreover, AIT was a safe treatment, with fever as the main adverse effects. This study adds to the evidence that postoperative HCC patients treated with adjuvant AIT show an improvement in disease-free survival rate or recurrence rate.     

A strategy for early detection of recurrent hepatocellular carcinoma following initial remission by transcatheter arterial chemoembolization

OBJECTIVES: To find a better surveillance method in detecting recurrent HCCs, patterns of recurrences following initial remission by transcatheter arterial chemoembolization (TACE) were evaluated. METHODS: Of 235 consecutive HCC patients who underwent TACE, 69 with initial remission were followed for >12 months. We compared the recurrence rates according to the characteristics of original HCCs and analyzed the locations of recurrent HCCs. We also evaluated the diagnostic efficacies of CT scan with serum AFP, angiography and Lipiodol CT scan in detecting recurrent HCCs. RESULTS: In 37 of 69, recurrent HCCs were detected after a median period of 17 months. Multinodular HCCs recurred more frequently than single-nodular HCCs. All of 5 patients with portal vein thrombosis recurred. Although 46% of recurrences were adjacent to original tumors, 62% were separated from them (8% at both). HCC with heterogeneous lipiodol uptake frequently recurred adjacent to original tumors. Only 18 of 37 recurrent HCCs were initially detected by serum AFP and CT scans; 17 by angiography, 2 only by lipiodol CT scan. CONCLUSIONS: Regular angiography may be valuable in detecting recurrent HCCs, especially in multinodular HCC. HCC with heterogeneous lipiodol uptake should be treated in combination with local ablation therapy.     

Low-dose, long-term, intermittent interferon-alpha-2b therapy after radical treatment by radiofrequency ablation delays clinical recurrence in patients with hepatitis C virus-related hepatocellular carcinoma

OBJECTIVE: To assess whether or not interferon (IFN) therapy prevents recurrence, and eventually improves the prognosis of patients with hepatocellular carcinoma (HCC) after completion of radical radiofrequency ablation (RFA) therapy. METHODS: Included as the IFN group in this study were 24 patients in total, who received radical RFA therapy first, followed by medication with IFN-alpha2b at such a low dose of 3 MIU x 2/week for as long as possible. On the other hand, the control group comprised 33 patients in total, who received radical RFA therapy without subsequent treatment with IFN. The control group was matched to the IFN group in age, platelet counts and size of nodules. RESULTS: Of the 24 patients treated with IFN, only one patient showed sustained virologic response. The median tumor-free period until the first recurrence after radical RFA therapy was 3.4 years in the IFN group and 1.4 years in the control group (p = 0.02). During the first 3 years after commencement of IFN administration, the cumulative recurrence rate in the IFN group was found to be lower than in the control group (p = 0.01); however, with the lapse of time over 3 years, the recurrence rate in the IFN group increased. There was no difference in the cumulative survival rates between the IFN group and the control group (p = 0.25). CONCLUSION: Subsequently after radical RFA therapy, long-term, low-dose, intermittent IFN therapy successfully delayed clinical recurrence of HCC.     

Successful laparoscopic extirpation of peritoneal dissemination after hepatectomy for ruptured hepatocellular carcinoma

Clinically, peritoneal dissemination of hepatocellular carcinoma (HCC) rarely occurs. We herein report a case that had a good outcome following laparoscopic extirpation of peritoneal dissemination after hepatectomy for ruptured HCC. A 66‐year‐old man underwent central bisectionectomy 12 days after emergency transcatheter arterial embolization for a ruptured HCC. Thereafter, pulmonary resection was performed twice for lung metastasis. About 8 months after the second pulmonary resection, a mass lesion was detected at the left subphrenic space on CT and ¹⁸F‐fluorodeoxyglucose PET scans. We made a diagnosis of peritoneal dissemination of HCC, and laparoscopic extirpation was performed. The patient is now doing well without any signs of recurrence 2 years after the last operation. Laparoscopic surgical resection for peritoneal dissemination that develops after hepatectomy for HCC may have a beneficial effect as a less‐invasive approach and may improve the prognosis in select patients.     

Serum alpha 1-antitrypsin in patients with hepatocellular carcinoma.

To evaluate the diagnostic value of alpha 1-antitrypsin (alpha-AT) as a tumor marker for hepatocellular carcinoma (HCC), we studied the serum levels of alpha-AT by rocket immunoelectrophoresis and alpha-fetoprotein (alpha-FP) by radioimmunoassay in 46 proven HCC patients, 43 cirrhosis patients and 200 healthy blood donors. The mean alpha-AT level of the 46 patients with HCC (4.8 +/- 2.7 mg/ml) was significantly higher than that of 200 healthy control subjects (1.7 +/- 0.7 mg/ml) (P less than 0.0001). The sensitivity of alpha-AT in 24 patients with high level of alpha-FP (greater than 400 ng/ml) and 22 patients with low level of alpha-FP (less than 400 ng/ml) were 96% and 64%, respectively. There was no substantial correlation between alpha-FP and alpha-AT in the two groups (alpha-FP greater than 400 ng/ml, alpha-FP less than 400 ng/ml) (r = 0.078, 0.064). The sensitivity for HCC using alpha-FP level alone (greater than 400 ng/ml) was only 52%, and the sensitivity using alpha-AT level alone (greater than 3.2 mg/ml) was 76% of the 46 patients. Combining both tests, sensitivity was improved only to 80%.     

Comprehensive proteomic profiling identifies serum proteomic signatures for detection of hepatocellular carcinoma and its subtypes

BACKGROUND: Detection of hepatocellular carcinoma (HCC) in patients with chronic liver disease (CLD) is difficult. We investigated the use of comprehensive proteomic profiling of sera to differentiate HCC from CLD. METHODS: Proteomes in sera from 20 CLD patients with alpha-fetoprotein (AFP) <500 microg/L (control group) and 38 HCC patients (disease group) were profiled by anion-exchange fractionation (first dimension), two types (IMAC3 copper and WCX2) of ProteinChip Arrays (second dimension), and time-of-flight mass spectrometry (third dimension). Bioinformatic tests were used to identify tumor-specific proteomic features and to estimate the values of the tumor-specific proteomic features in the diagnosis of HCC. Cross-validation was performed, and we also validated the models with pooled sera from the control and disease groups, serum from a CLD patient with AFP >500 microg/L, and postoperative sera from two HCC patients. RESULTS: Among 2384 common serum proteomic features, 250 were significantly different between the HCC and CLD cases. Two-way hierarchical clustering differentiated HCC and CLD cases. Most HCC cases with advanced disease were clustered together and formed two subgroups that contained significantly more cases with lymph node invasion or distant metastasis. For differentiation of HCC and CLD by an artificial network (ANN), the area under the ROC curve was 0.91 (95% confidence interval, 0.82-1.01; P <0.0005) for all cases and 0.954 (95% confidence interval, 0.881-1.027; P <0.0005) for cases with nondiagnostic serum AFP (<500 microg/L). At a specificity of 90%, the sensitivity was 92%. Both cluster analysis and ANN correctly classified the pooled serum samples, the CLD serum sample with increased AFP, and the HCC patient in complete remission. CONCLUSION: Tumor-specific proteomic signatures may be useful for detection and classification of hepatocellular cancers.     

Minimally invasive treatment combined with cytokine-induced killer cells therapy lower the short-term recurrence rates of hepatocellular carcinomas

The recurrence of hepatocellular carcinoma (HCC) after minimally invasive therapy is frequent. Adoptive immunotherapy is thought to be an effective method to lower recurrence and metastasis rates of malignant tumors. Therefore, 85 HCC patients after transcatheter arterial chemoembolization and radiofrequency ablation therapy were randomized to immunotherapy group and no adjuvant therapy group. Autologous cytokine-induced killer (CIK) cells were transfused via hepatic artery to the patients. The alteration of levels of lymphocyte subsets in peripheral blood of patients was examined by flow cytometry. All patients were screened by computed tomography every 2 months to observe the tumor recurrent conditions. After CIK cell infusions, the percentages of CD3+, CD4+, CD56+, CD3+CD56+ cells, and CD4+/CD8+ ratio increased from 68.6+/-11.0%, 31.1+/-9.0%, 15.6+/-7.9%, 5.2+/-3.1%, and 1.1+/-0.5 to 70.7+/-10.1%, 33.5+/-8.0%, 18.4+/-9.4%, 5.9+/-2.8%, and 1.3+/-0.7, respectively (P<0.05); whereas the percentage of CD8 cells decreased from 31.1+/-7.8% to 28.6+/-8.3% (P<0.05). The 1-year and 18-month recurrence rates of the study group were 8.9% and 15.6%, compared with 30.0% and 40.0% of the control group (both P value<0.05). The data suggest that CIK cell transfusion is an effective treatment. It can boost the immunologic function in HCC patients and plays an important role in reducing the recurrence rate of HCC.