Nomogram prediction of vessels encapsulating tumor clusters in small hepatocellular carcinoma≤3 cm based on enhanced magnetic resonance imaging

BACKGROUND Vessels encapsulating tumor clusters(VETC)represent a recently discovered vascular pattern associated with novel metastasis mechanisms in hepatocellular carcinoma(HCC).However,it seems that no one have focused on predicting VETC status in small HCC(sHCC).This study aimed to develop a new nomogram for predicting VETC positivity using preoperative clinical data and image features in sHCC(≤3 cm)patients.AIM To construct a nomogram that combines preoperative clinical parameters and image features to predict patterns of VETC and evaluate the prognosis of sHCC patients.METHODS A total of 309 patients with sHCC,who underwent segmental resection and had their VETC status confirmed,were included in the study.These patients were recruited from three different hospitals:Hospital 1 contributed 177 patients for the training set,Hospital 2 provided 78 patients for the test set,and Hospital 3 provided 54 patients for the validation set.Independent predictors of VETC were identified through univariate and multivariate logistic analyses.These independent predictors were then used to construct a VETC prediction model for sHCC.The model’s performance was evaluated using the area under the curve(AUC),calibration curve,and clinical decision curve.Additionally,Kaplan-Meier survival analysis was performed to confirm whether the predicted VETC status by the model is associated with early recurrence,just as it is with the actual VETC status and early recurrence.RESULTS Alpha-fetoprotein_lg10,carbohydrate antigen 199,irregular shape,non-smooth margin,and arterial peritumoral enhancement were identified as independent predictors of VETC.The model incorporating these predictors demonstrated strong predictive performance.The AUC was 0.811 for the training set,0.800 for the test set,and 0.791 for the validation set.The calibration curve indicated that the predicted probability was consistent with the actual VETC status in all three sets.Furthermore,the decision curve analysis demonstrated the clinical benefits of our model for patients with sHCC.Finally,early recurrence was more likely to occur in the VETC-positive group compared to the VETC-negative group,regardless of whether considering the actual or predicted VETC status.CONCLUSION Our novel prediction model demonstrates strong performance in predicting VETC positivity in sHCC(≤3 cm)patients,and it holds potential for predicting early recurrence.This model equips clinicians with valuable information to make informed clinical treatment decisions.

Small hepatocellular carcinoma: current status and prospects

Background: More than two decades have gone by since the early report of resection for small hepatocel- lular carcinoma (HCC), which resulted in improved prognosis of HCC. Objective: To review the past and recent data, and prospect the future in this field. Data sources: Literature and recent data from the Liver Cancer Institute of Fudan University, Shang- hai, China. Data synthesis: 1232 patients with small HCC from the institute were analyzed between 1960-1984 (n= 107) and 1985-1999 (n=1125). The increase of li- mited resection rate from 69.5 % to 82.5 % contribu- ted in part to the increase of resectability from 76.6 % to 95.5 %, decrease of operative mortality from 2.4 % to 1.2 %, and improvement of 5-year sur- vival after resection (from 53.1% to 64.0%). The 5-year survival was higher after limited resection than after lobectomy, being 64.4 % versus 55.9%. The 5-year survival after resection was superior to that after cryosurgery and other regional cancer therapies (32.8 %). However, molecular studies found that biological characteristics were only slightly better in small HCC than in large HCC. Conclusions: Resection remains the treatment choice for small HCC with compensated liver function, while regional cancer therapies and liver transplanta- tion are alternatives for patients with incompensated liver function. Biological characteristics remain the leading factor influencing prognosis of small HCC.

Therapeutic potential of small interfering RNAs/micro interfering RNA in hepatocellular carcinoma

Hepatocellular carcinoma(HCC) is the predominant form of primary liver cancer and represents the third leading cause of cancer-related death worldwide. Current available therapeutic approaches are poorly effective,especially for the advanced forms of the disease. In the last year,short double stranded RNA molecules termed small interfering RNAs(si RNAs) and micro interfering RNAs(mi RNA),emerged as interesting molecules with potential therapeutic value for HCC. The practical use of these molecules is however limited by the identification of optimal molecular targets and especially by the lack of effective and targeted HCC delivery systems. Here we focus our discussion on the most recent advances in the identification of si RNAs/mi RNAs molecular targets and on the development of suitable si RNA/mi RNAs delivery systems.

Survival factors after resection of small hepatocellular carcinoma

Early resection of hepatocellular carcinoma is a key measure to prolong the survival of patients. This study was designed to summarize our experience in surgical resection of small hepatocellular carcinoma (HCC), and to analyze the factors influencing the postoperative survival of patients. METHODS:The clinicopathologic data of 105 patients with small HCC after resection from 1986 through 2003 were analyzed ; the patients had been followed up for more than half a year (median 33 months). Nine clinicopathologic factors, preoperative α-fetoprotein (AFP) level, liver cirrhosis, Child-Pugh score, tumor size (>2cm vs.≤2 cm) and number (single vs. multiple), capsule formation, portal vein tumor thrombi (PVTT), Edmondson tumor grade and surgical method, were analyzed by the log-rank test and the Cox proportional harzards model analysis. RESULTS:The cumulative 1-,3- and 5-year survival rates after the operation were 86.5%, 70.3% and 55.2%, respectively, and the 1-, 3- and 5-year disease-free survival rates were 78%, 58. 9% and 45. 6%, respectively. One patient died from esophagogastric varices hemorrhage in 2 weeks after reoperation. Thirty-six patients had intrahepatic recurrence or metastasis postoperatively and 34 patients died. The Kaplan-Meier method and the Cox proportional harzards model analysis indicated that poor Child-Pugh score, tumor more than 2 cm in diameter, PVTT and multiple lesions (including satellitic lesions) were adverse factors affecting postoperative survival. The Cox proportional harzards model analysis indicated that tumor size, PVTT and multiple lesions were the factors influencing postoperative disease-free survival. CONCLUSIONS:Limited hepatectomy with a margin more than 1 cm is an appropriate surgical approach. Adverse preoperative Child-Pugh score and postoperative intrahepatic recurrences are the main factors leading to the death of patients with small HCC.

Des-gamma-carboxy prothrombin as an important prognostic indicator in patients with small hepatocellular carcinoma

AIM:To clarify the effect of a high des-gamma-carboxy prothrombin (DCP) level on the invasiveness and prognosis of small hepatocellular carcinoma. METHODS: Among 142 consecutive patients with known DCP levels, who underwent hepatectomy because of hepatocellular carcinoma, 85 patients met the criteria for small hepatocellular carcinoma, i.e. one ≤ 5 cm sized single tumor or no more than three ≤ 3 cm sized tumors. RESULTS: The overall survival rate of the 142 patients was 92.1% for 1 year, 69.6% for 3 years, and 56.9% for 5 years. Multivariate analysis showed that microscopic vascular invasion (P = 0.03) and serum DCP ≥ 400 mAU/mL (P = 0.02) were independent prognostic factors. In the group of patients who met the criteria for small hepatocellular carcinoma, DCP ≥ 400 mAU/mL was found to be an independent prognostic factor for recurrence-free (P = 0.02) and overall survival (P = 0.0005). In patients who did not meet the criteria, the presence of vascular invasion was an independent factor for recurrence-free (P = 0.02) and overall survivals (P = 0.01). In 75% of patients with small hepatocellular carcinoma and high DCP levels, recurrence occurred extrahepatically. CONCLUSION: For small hepatocellular carcinoma, a high preoperative DCP level appears indicative fortumor recurrence. Because many patients with a high preoperative DCP level develop extrahepatic recurrence, it is necessary to screen the whole body.

Prognostic factors affecting postoperative survival of patients with solitary small hepatocellular carcinoma

Background: Small hepatocellular carcinoma(sHCC) is a unique variant of HCC that is characterized by small tumor size(maximum tumor diameter predic≤3 cm) and favorable long?term outcomes. The present study aimed to define clin?icopathologic factors that t survival in patients with s HCC.Methods: The study population consisted of 335 patients who underwent hepatectomy for solitary s HCC between December 1998 and 2010. Prognostic factors were evaluated using Kaplan–Meier curves and Cox proportional hazard models.Results: The 5?year overall survival(OS) and recurrence?free survival(RFS) rates were 77.7% and 59.9%, respectively. Kaplan–Meier curves showed that tumor size and vascular invasion had prognostic significance within this relatively selected cohort(P < 0.05). Multivariate analysis confirmed that increased tumor size and vascular invasion were independent prognostic factors for short OS(hazard ratio [HR] = 2.367, 95% confidence interval [CI] 1.406–3.985; HR = 2.954, 95% CI 1.781–4.900) and RFS(HR = 1.779, 95% CI 1.259–2.514; HR < 0.05). Importantly, a proposed prognostic scoring model was deri= 1.699, 95% CI 1.165–2.477) in s HCC patients(Pved according to the two variables; tumor size and extent of vascular invasion were significantly associated with OS and RFS in patients with s HCC(P < 0.001).Conclusions: Tumor size and vascular invasion are feasible and useful prognostic factors for s HCC. The proposed prognostic model, based on tumor size and vascular invasion, is informative in predicting survival in s HCC patients undergoing hepatectomy.

A special recurrent pattern in small hepatocellular carcinoma after treatment:Bile duct tumor thrombus formation

AIM:To investigate the clinicopathologic features of bile duct tumor thrombus(BDTT) occurrence after treatment of primary small hepatocellular carcinoma(sHCC) .METHODS:A total of 423 patients with primary sHCC admitted to our hospital underwent surgical resection or local ablation.During follow-up,only six patients were hospitalized due to obstructive jaundice,which occurred 5-76 mo after initial treatment.The clinicopathologic features of these six patients were reviewed.RESULTS:Six patients underwent hepatic resection(n=5) or radio-frequency ablation(n=1) due to primary sHCC.Five cases had an R1 resection margin,and one case had an ablative margin less than 5.0 mm.No vascular infiltration,microsatellites or bile duct/canaliculus affection was noted in the initial resected specimens.During the follow-up,imaging studies revealed a macroscopic BDTT extending to the common bile duct in all six patients.Four patients had a concomitant intrahepatic recurrent tumor.Surgical re-resection of intrahepatic recurrent tumors and removal of BDTTs(n=4) ,BDTT removal through choledochotomy(n= 1) ,and conservative treatment(n=1) was performed.Microscopic portal vein invasion was noted in three of the four resected specimens.All six patients died,with a mean survival of 11 mo after BDTT removal or conservative treatment.CONCLUSION:BDTT occurrence is a rare,special recurrent pattern of primary sHCC.Patients with BDTTs extending to the common bile duct usually have an unfavorable prognosis even following aggressive surgery.Insufficient resection or ablative margins against primary sHCC may be a risk factor for BDTT development.

Comparison of unenhanced magnetic resonance imaging and ultrasound in detecting very small hepatocellular carcinoma

BACKGROUND In hepatocellular carcinoma(HCC),detection and treatment prior to growth beyond 2 cm are important as a larger tumor size is more frequently associated with microvascular invasion and/or satellites.In the surveillance of very small HCC nodules(≤2 cm in maximum diameter,Barcelona clinical stage 0),we demonstrated that the tumor markers alpha-fetoprotein and PIVKA-Ⅱare not so useful.Therefore,we must survey with imaging modalities.The superiority of magnetic resonance imaging(MRI)over ultrasound(US)to detect HCC was confirmed in many studies.Although enhanced MRI is now performed to accurately diagnose HCC,in conventional clinical practice for HCC surveillance in liver diseases,unenhanced MRI is widely performed throughout the world.While,MRI has made marked improvements in recent years.AIM To make a comparison of unenhanced MRI and US in detecting very small HCC that was examined in the last ten years in patients in whom MRI and US examinations were performed nearly simultaneously.METHODS In 394 patients with very small HCC nodules,those who underwent MRI and US at nearly the same time(on the same day whenever possible or at least within 14 days of one another)at the first diagnosis of HCC were selected.The detection rate of HCC with unenhanced MRI was investigated and compared with that of unenhanced US.RESULTS The sensitivity of unenhanced MRI for detecting very small HCC was 95.1%(97/102,95%confidence interval:90.9-99.3)and that of unenhanced US was 69.6%(71/102,95%confidence interval:60.7-78.5).The sensitivity of unenhanced MRI for detecting very small HCC was significantly higher than that of unenhanced US(P<0.001).Regarding the location of HCC in the liver in patients in whom detection by US was unsuccessful,S7-8 was identified in 51.7%.CONCLUSION Currently,unenhanced MRI is a very useful tool for the surveillance of very small HCC in conventional clinical follow-up practice.

Intussusception due to hematogenous metastasis of hepatocellular carcinoma to the small intestine:A case report

BACKGROUND The commonest sites of extrahepatic metastases from hepatocellular carcinoma(HCC)are the lungs,bones,adrenal glands,and regional lymph nodes.Hematogenous metastasis to the gastrointestinal(GI)tract is a rare condition in patients with HCC,and the prognosis is usually poor.We report,herein,an extremely rare case of a patient with intussusception due to hematogenous metastasis of HCC to the ileum and his long-term survival with multidisciplinary therapy.CASE SUMMARY The patient was a 71-year-old man with a history of chronic hepatitis B,who had undergone three surgeries for HCC.He was treated with sorafenib for peritoneal metastases of HCC.He was admitted to our hospital with chief complaints of abdominal pain and vomiting.Abdominal contrast-enhanced computed tomography imaging revealed a small intestinal tumor,presenting with intussusception and small bowel obstruction.Conservative treatment was started,but due to repeated exacerbation of symptoms,surgery was planned on the 28th d of hospitalization.Partial ileal resection without reducing the intussusception and end-to-end anastomosis was performed.On histological examination,tumor cells were not observed on the serosal surface,but intravascular invasion of tumor cells was seen.Immunohistochemistry was positive for immunohistochemical markers,and a diagnosis of hematogenous metastasis of HCC to the ileum was made.He remains alive 82 mo after the first surgery.CONCLUSION Prognosis of HCC patients with GI tract metastasis is usually poor,but in some cases,multidisciplinary therapy may prolong survival.

THE USEFULNESS OF ^(99)Tc-PMT DELAYED HEPATOBILIARY IMAGING IN THE DIAGNOSIS OF SMALL HEPATOCELLULAR CARCINOMA

This investigation was aimed to assess the usefulness of delayed hepalobillary Imaging in the diagnosis of small hepatocellular carcinoma (HCC). Sixty-two patients with this hepatic cancer were Included in the study. 56 males and 6 females, with a mean age of 50. 6 yr. (32 - 72 years old). All patients were performed by surgery, verified histologically, and these tumors were smaller than 5 cm. Liver scans were performed 5 minutes, 2 hours and 5 hours after the administration of radlopharmaceutices. In 31 of the 62 patients (50%), the tumor exhibited equal radioactivity uptake or greater radioactivity uptake than the surrounding liver in delayed imaging. And the sensitivity was 33. 3% (2/6), 41.2% (7/17), 60.0% (9/15) and 54.2% (13/24) In the tumor size was ≤2 cm, 2-3cm, 3-4 cm and 4 - 5 cm, respectively. The smallest mass to be detected was only 1. 2 cm. The uptake of radiopharmaceutic was nonsignificantly related to serum AFP level and hepatic cirrhosis (P>0. 05). These results show that 99-Tc-PMT delayed hepatobiliary imaging can be useful in the diagnosis of small hepatocellular carcinoma.

Small-duct primary sclerosing cholangitis with hepatocellular carcinoma requiring liver transplantation

BACKGROUND:Primary sclerosing cholangitis(PSC)is a chronic progressive cholestatic liver disease,which usually affects young adults and is diagnosed by cholangiography.On a few occasions,the disease either starts in or exclusively involves the small intrahepatic bile ducts,referred to as small-duct PSC. METHODS:A 31-year-old man presented with severe hematemesis secondary to liver cirrhosis.Over a course of 8 years,his liver decompensated and required an orthotopic liver transplantation. In this report we discuss his disease presentation,course of management,and the post-transplantation course of manage- ment,and review the morphologic diagnosis,and differential diagnosis of the disease with large-duct type and other diseases that involve small intrahepatic bile ducts. RESULTS:The patient’s explanted liver showed changes of PSC affecting only the small-and medium-sized bile ducts in addition to three incidental nodules of hepatocellular carcinoma. CONCLUSIONS:Small-duct PSC has a substantially better prognosis than the large-duct type,with less chance of developing cirrhosis and an equal risk for developing hepato- cellular carcinoma,but no increased risk for developing cholangiocarcinoma.Treatment seems to help relieve the symptoms but not necessarily improve survival.Liver transplantation remains the ultimate cure.

Correlation of small hepatocellular carcinoma ultrasonography parameters with the expression of oncogenes and angiogenesis genes

Objective: To investigate the correlation of small hepatocellular carcinoma ultrasonography parameters with the expression of oncogenes and angiogenesis genes. Methods: A total of 61 patients with small hepatocellular carcinoma who were diagnosed in this hospital between July 2015 and May 2017 were selected as small hepatocellular carcinoma group, and 48 patients with hepatolithiasis were selected as hepatolithiasis group. The ultrasonography parameters of the two groups were recorded and the expression levels of oncogenes and angiogenesis genes in the surgical lesion tissues were detected. Pearson test was used to evaluate the correlation of ultrasonography parameters with the expression of oncogenes and angiogenesis genes. Results: IMAX level in small hepatocellular carcinoma group was higher than that in hepatolithiasis group while TTP and mTT levels were lower than those in hepatolithiasis group;oncogenes C-myc, N-ras, PIK3CA, RMP, Bmil and pim-3 mRNA expression in lesion tissues were higher than those of hepatolithiasis group;angiogenesis genes VEGF, Ang-1, Tie-2 and MACC1 mRNA expression in lesion tissues were higher than those of hepatolithiasis group while ARH1 mRNA expression was lower than that of hepatolithiasis group. Pearson test showed that the ultrasonography parameters IMAX, TTP and mTT levels in patients with small hepatocellular carcinoma were directly correlated to the expression of oncogenes and angiogenesis genes in lesions. Conclusion: The ultrasonography parameters of patients with small hepatocellular carcinoma are significantly different from those of patients with benign diseases, and the specific parameter levels are directly correlated with the malignancy of cancer cells.

NHE7 upregulation potentiates the uptake of small extracellular vesicles by enhancing maturation of macropinosomes in hepatocellular carcinoma

Background:Small extracellular vesicles(sEVs)mediate intercellular commu-nication that contributes to hepatocellular carcinoma(HCC)progression via multifaceted pathways.The success of cell entry determines the effect of sEV on recipient cells.Here,we aimed to delineate the mechanisms underlying the uptake of sEV in HCC.Abbreviations:AF,Alexa Fluor;ANOVA,analysis of variance;ATP9A,ATPase Phospholipid Transporting 9A;BCECF-AM,2’,7’-bis-(2-barboxyethyl)-5-(and-6)-carboxyfluorescein,acetoxymethyl ester;BSA,bovine serum albumin;CCMR,Centre for Comparative Medicine Research;CRISPR,clustered regularly interspaced short palindromic repeats;CTL,ctrl;CXCR4,C-X-C Chemokine Receptor Type 4;DAPI,4′,6-diamidino-2-phenylindole;DFS,disease-free survival;DMEM,Dulbecco’s Modified Eagle Medium;DMSO,dimethyl sulfoxide;Dox,doxycycline;EEA1,early endosome antigen 1;EIPA,5-(N-ethyl-N-isopropyl)-amiloride;FBS,fetal bovine serum;FITC,fluorescein isothiocyanate;GAPDH,glyceraldehyde-3-phosphate dehydrogenase;GM130,Golgi matrix protein 130;HCC,hepatocellular carcinoma;HEPES,4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid;HPRT1,hypoxanthine phosphoribosyltransferase 1;H-score,histoscore;IAA,indole-3-acetic acid;KD,knockdown;KO,knockout;mAID,mini-auxin-inducible degron;MTT,3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide;NHE7,Na(+)/H(+)exchanger 7;ns,non-significant;OD,optical density;OS,overall survival;PBS,phosphate-buffered saline;PCR,polymerase chain reaction;pHe,endosomal pH;pHi,intracellular pH;PKH67,Paul Karl Horan 67;Rab21,Ras-associated binding protein 21;RIPA,radioimmunoprecipitation assay;SAM,synergistic activation mediator;SEMs,standard error of the means;sEVs,small extracellular vesicles;sgRNA,single-guide RNA;shRNA,short-hairpin RNA;SLC9,solute carrier gene 9;SLiCE,Seamless Ligation Cloning Extract;TCGA,The Cancer Genome Atlas;TCL,total cell lysates;TGN,trans-Golgi network;TMA,tissue microarray;TMR,tetramethyl rhodamine;TSG101,tumor susceptibility gene 101.Yue Yao and Yi Xu contributed equally to this work.Methods:Macropinocytosis was examined by the ability of cells to internalize dextran and sEV.Macropinocytosis was analyzed in Na(+)/H(+)exchanger 7(NHE7)-knockdown and-overexpressing cells.The properties of cells were stud-ied using functional assays.pH biosensor was used to evaluate the intracellular and endosomal pH.The expression of NHE7 in patients’liver tissues was exam-ined by immunofluorescent staining.Inducible silencing of NHE7 in established tumors was performed to reveal the therapeutic potential of targeting NHE7.Results:The data revealed that macropinocytosis controlled the internaliza-tion of sEVs and their oncogenic effect on recipient cells.It was found that metastatic HCC cells exhibited the highest efficiency of sEV uptake relative to normal liver cells and non-metastatic HCC cells.Attenuation of macropinocytic activity by 5-(N-ethyl-N-isopropyl)-amiloride(EIPA)limited the entry of sEVs and compromised cell aggressiveness.Mechanistically,we delineated that high level of NHE7,a sodium-hydrogen exchanger,alkalized intracellular pH and acidized endosomal pH,leading to the maturation of macropinosomes.Inducible inhibition of NHE7 in established tumors developed in mice delayed tumor development and suppressed lung metastasis.Clinically,NHE7 expression was upregulated and linked to dismal prognosis of HCC.Conclusions:This study advances the understanding that NHE7 enhances sEV uptake by macropinocytosis to promote the malignant properties of HCC cells.Inhibition of sEV uptake via macropinocytosis can be exploited as a treatment alone or in combination with conventional therapeutic approaches for HCC.

Patients with early recurrence of hepatocellular carcinoma have poor prognosis

BACKGROUND： Early recurrence （ER） after hepatic resection （HR） is a poor prognostic factor for patients with hepatocellular carcinoma （HCC）. This study aimed to identify the clinico- pathological features, outcomes, and risk factors for ER after HR for small HCC in order to clarify the reasons why ER is a worse recurrence pattern. METHODS： We retrospectively examined 130 patients who underwent HR for small HCC （___30 mm）. Recurrence was clas- sifted into ER （〈2 years） and late recurrence （LR） （_〉2 years）. The clinicopathological features, outcomes, and risk factors for ER were analyzed by multivariate analysis. RESULTS： ER was observed in 39 patients （30.0%）. The sur- vival rate of the ER group was significantly lower than that of the LR group （P〈0.005）, and ER was an independent prognos- tic factor for poor survival （P=0.0001）. The ER group had a significantly higher frequency （P=0.0039） and shorter interval （P=0.027） of development to carcinoma beyond the Milan criteria （DBMC） compared with the LR group, and ER was an independent risk factor for DBMC （P〈0.0001）. Multi-nodularity, non-simple nodular type, and microvascular invasion were independent predictors for ER （P=0.012, 0.010, and 0.019, respectively）.CONCLUSIONS： ER was a highly malignant recurrence pattern associated with DBMC and subsequent poor survival after HR for small HCC. Multi-nodularity, non-simple nodular type, and microvascular invasion predict ER, and taking these factors into consideration may be useful for the decision of the treatment strategy for small HCC after HR.

Latest developments in precancerous lesions of hepatocellular carcinoma

Hepatocarcinogenesis in human chronic liver diseases is a multi-step process in which hepatic precancerous lesions progress into early hepatocellular carcinoma(HCC) and progressed HCC, and the close surveillance and treatment of these lesions will help improve the survival rates of patients with HCC. The rapid development and extensive application of imaging technology have facilitated the discovery of nodular lesions of ambiguous significance, such as dysplastic nodules. Further investigations showed that these nodules may be hepatic precancerous lesions, and they often appear in patients with liver cirrhosis. Although the morphology of these nodules is not sufficient to support a diagnosis of malignant tumor, these nodules are closely correlated with the occurrence of HCC, as indicated by long-term follow-up studies. In recent years, the rapid development and wide application of pathology, molecular genetics and imaging technology have elucidated the characteristics of precancerous lesions. Based on our extensive review of the relevant literature, this article focuses on evidence indicating that high-grade dysplastic nodules are more likely to transform into HCC than low-grade dysplastic nodules based on clinical, pathological, molecular genetic and radiological assessments. In addition, evidence supporting the precancerous nature of large cell change in hepatitis B virus-related HCC is discussed.

Single hepatocellular carcinoma ≤ 3 cm in left lateral segment:Liver resection or radiofrequency ablation?

AIM: To evaluate the long-term results of radiofrequency ablation (RFA) compared to left lateral sectionectomy (LLS) in patients with Child-Pugh class A disease for the treatment of single and small hepatocellular carcinoma (HCC) in the left lateral segments.

A potential oncogenic role of the commonly observed E2F5 overexpression in hepatocellular carcinoma

AIM: To explore the expression pattern of E2F5 in primary hepatocellular carcinomas (HCCs) and elucidate the roles of E2F5 in hepatocarcinogenesis. METHODS: E2F5 expression was analyzed in 120 primary HCCs and 29 normal liver tissues by immunohistochemistry analysis. E2F5-small interfering RNA was transfected into HepG2, an E2F5-overexpressed HCC cell line. After E2F5 knockdown, cell growth capacity and migrating potential were examined. RESULTS: E2F5 was significantly overexpressed in primary HCCs compared with normal liver tissues (P = 0.008). The E2F5-silenced cells showed significantly reduced proliferation (P = 0.004). On the colony formation and soft agar assays, the number of colonies was significantly reduced in E2F5-silenced cells (P = 0.004 and P = 0.009, respectively). E2F5 knockdown resulted in the accumulation of G0/G1 phase cells and a reduction of S phase cells. The number of migrating/invading cells was also reduced after E2F5 knockdown (P = 0.021). CONCLUSION: To our knowledge, this is the first evidence that E2F5 is commonly overexpressed in primary HCC and that E2F5 knockdown significantly repressed the growth of HCC cells.

GABA stimulates human hepatocellular carcinoma growth through overexpressed GABAA receptor theta subunit

AIM: To investigate the function of gamma-aminobutyric acid (GABA) and gamma-aminobutyric acid A receptor θ subunit (GABRQ) in hepatocellular carcinoma (HCC). METHODS: Semiquantitative polymerase chain reaction was used for detecting the expression of GABRQ receptor among HCC cell line HepG2, normal liver cell line L-02, non-malignant Chang's liver cells, 8 samples of HCC tissues and paired non-cancerous tissues. HepG2 cells were treated with GABA at serial concentrations (0, 1, 10, 20, 40 and 60 μmol/L), and their proliferating abilities were analyzed with the methyl thiazolyl tetrazolium assay, cell cycle analysis and tumor implanted in nude mice. Small interfering RNA was used for knocking down the endogenous GABRQ in HepG2. Proliferating abilities of these cells treated with or without GABA were analyzed. RESULTS: We identified the overexpression of GABRQ in HCC cell lines and half of the tested HCC tissues. Knockdown of endogenous GABRQ expression in HepG2 attenuated HCC cell growth, suggesting its role in HCC cell viability. We studied the effect of GABA in the proliferation of GABRQ-positive cell lines in vitro and in vivo , and found that GABA increased HCC growth in a dosedependent manner. Notably, the addition of GABA into the cell culture medium promoted the proliferation of GABRQ-expressing HepG2 cells, but not GABRQ-knockdown HepG2 cells, which means that GABA stimulates HepG2 cell growth through GABRQ. CONCLUSION: GABRQ play important roles in HCC development and progression and could be a promising molecular target for the development of new diagnostic and therapeutic strategies of HCC.

The Role of Multimodality Imaging Techniques on Differential Diagnosis of Precancerous Nodules and Hepatocellular Carcinoma

In recent years, the incidence of hepatocellular carcinoma (HCC) has been increasing worldwide, and its high mortality seriously threatens public health. Early detection and treatment are crucial to improving the survival rate. Imaging examination widely used for the diagnosis of HCC and provides a non-invasive means of tumor visualization. The rapid development of medical imaging technology is expected to improve early-stage diagnosis rates for HCC. This article summarizes the methods for the differential diagnosis of premalignant dysplastic nodule (DN) and small hepatocellular carcinoma during the carcinogenesis of cirrhosis and reviews their application. In addition, a discussion on some recently patented medical imaging development was also presented.

Development and in vitro study of a bi-specific magnetic resonance imaging molecular probe for hepatocellular carcinoma

BACKGROUND Hepatocellular carcinoma(HCC)ranks second in terms of cancer mortality worldwide.Molecular magnetic resonance imaging(MRI)targeting HCC biomarkers such as alpha-fetoprotein(AFP)or glypican-3(GPC3)offers new strategies to enhance specificity and help early diagnosis of HCC.However,the existing iron oxide nanoparticle-based MR molecular probes singly target AFP or GPC3,which may hinder their efficiency to detect heterogeneous micro malignant HCC tumors<1 cm(MHCC).We hypothesized that the strategy of double antibody-conjugated iron oxide nanoparticles which simultaneously target AFP and GPC3 antigens may potentially be used to overcome the tumor heterogeneity and enhance the detection rate for MRI-based MHCC diagnosis.AIM To synthesize an AFP/GPC3 double antibody-labeled iron oxide MRI molecular probe and to assess its impact on MRI specificity and sensitivity at the cellular level.METHODS A double antigen-targeted MRI probe for MHCC anti-AFP-USPIO-anti-GPC3(UAG)was developed by simultaneously conjugating AFP andGPC3 antibodies to a 5 nm ultra-small superparamagnetic iron oxide nanoparticle(USPIO).At the same time,the singly labeled probes of anti-AFP-USPIO(UA)and anti-GPC3-USPIO(UG)and non-targeted USPIO(U)were also prepared for comparison.The physical characterization including morphology(transmission electron microscopy),hydrodynamic size,and zeta potential(dynamic light scattering)was conducted for each of the probes.The antigen targeting and MRI ability for these four kinds of USPIO probes were studied in the GPC3-expressing murine hepatoma cell line Hepa1-6/GPC3.First,AFP and GPC3 antigen expression in Hepa1-6/GPC3 cells was confirmed by flow cytometry and immunocytochemistry.Then,the cellular uptake of USPIO probes was investigated by Prussian blue staining assay and in vitro MRI(T2-weighted and T2-map)with a 3.0 Tesla clinical MR scanner.RESULTS Our data showed that the double antibody-conjugated probe UAG had the best specificity in targeting Hepa1-6/GPC3 cells expressing AFP and GPC3 antigens compared with single antibody-conjugated and unconjugated USPIO probes.The iron Prussian blue staining and quantitative T2-map MRI analysis showed that,compared with UA,UG,and U,the uptake of double antigen-targeted UAG probe demonstrated a 23.3%(vs UA),15.4%(vs UG),and 57.3%(vs U)increased Prussian stained cell percentage and a 14.93%(vs UA),9.38%(vs UG),and 15.3%(vs U)reduction of T2 relaxation time,respectively.Such bi-specific probe might have the potential to overcome tumor heterogeneity.Meanwhile,the coupling of two antibodies did not influence the magnetic performance of USPIO,and the relatively small hydrodynamic size(59.60±1.87 nm)of double antibodyconjugated USPIO probe makes it a viable candidate for use in MHCC MRI in vivo,as they are slowly phagocytosed by macrophages.CONCLUSION The bi-specific probe presents enhanced targeting efficiency and MRI sensitivity to HCC cells than singly-or non-targeted USPIO,paving the way for in vivo translation to further evaluate its clinical potential.