[Analysis of clinicopathological features of 18 cases of hepatic angiosarcoma].

Objective: To investigate the clinicopathological features, treatment, and prognosis of hepatic angiosarcoma. Methods: Clinicopathological data and prognostic conditions of 18 cases with hepatic angiosarcoma were collected retrospectively. The recurrence-free survival rate and overall survival rate were calculated by the Kaplan-Meier method. A Cox regression analysis was used to explore the survival-related risk factors. Results: There were 12 male and 6 female patients, with an average age of 57 (37 ~ 70) years. The tumor's average diameter was 8.40 (2.00 ~ 18.00) cm. Seven cases had multiple tumors, while two cases had large vessel tumor thrombuses. Microscopically, the tumor tissues were irregularly anastomosed, with vascular lacunar or solid bundle-like weaving, and the tissue morphology mimicked capillary hemangioma, cavernous hemangioma, or angioepithelioma, while tumor cells were spindle-shaped or epithelioid, lined with hobnails in the lumen, or formed papillary structures in the lumen. The proportion of highly, moderately, and poorly differentiated tumors was 4:8:6, with six cases having clear tumor boundaries, eight having microvascular tumor thrombi, and sixteen having blood lake formation. Different levels of expression of CD31, CD34, erythroblast transformation-specific related genes, and Fli-1 markers were demonstrated in all of the cases. Four cases had a P53 mutation, and six cases had Ki-67 > 10%. During the follow-up period of 0.23-114.20 months, the five-year recurrence-free survival rate and overall survival rate were 16.7% and 37.2%, respectively. Cox regression multivariate analysis showed that preoperative symptoms and multiple tumors were significant risk factors for recurrence-free survival, while preoperative symptoms and Ki-67 > 10% were significant risk factors for overall survival. Conclusion: Hepatic angiosarcoma is a rare hepatic mesenchymal tumor with high malignancy and a poor prognosis. Pathological morphology and immunohistochemical marker combinations are needed for a definite diagnosis. However, the complexity of angiosarcomas' histological and cytological conformations and the overlap of pathological features with benign vascular tumors, sarcomas, and carcinomas pose difficulties in the differential diagnosis. Thus, the only effective ways to prolong survival are early detection and radical surgical resection.

[Advances in the diagnostic pathological features of precancerous lesions of hepatocellular carcinoma].

The occurrence of hepatocellular carcinoma (HCC) is a multistep development process through precancerous lesions. A precancerous lesion of HCC is classified into hepatocyte dysplasia at the cytological level and dysplastic nodules at the histological level, and the corresponding lesion subtypes have different risks of canceration. Pathology is the "gold standard" for the diagnosis of early stage HCC and its precancerous lesions. However, it also faces many difficulties and challenges, such as the accumulation of experience in the pathological diagnosis, the understanding and grasp of key points of histopathological diagnosis and differential diagnosis, the combination application of immune and molecular diagnostic markers, and many others. This article briefly discusses the key points of pathological features and differential diagnosis of precancerous lesions of HCC.

[Copy number aberrations of genes related to extrohepatic metastasis-free survival after operation for hepatocellular carcinoma].

Objective: To investigate the molecular markers of copy number aberrations (CNAs) of genes related to extrohepatic metastasis-free survival after the operation for hepatocellular carcinoma (HCC). Methods: The CNA status of 20 candidate genes in 66 HCC samples was detected by microarray comparative genomic hybridization. The associations between gene CNAs and extrohepatic metastasis-free survival were evaluated using the Cox regression model, Log-rank test, and Kaplan-Meier survival analysis. Results: Multivariate Cox analysis revealed that the independent risk factors for metastasis-free survival were MDM4 gain (hazard ratio [HR] = 2.74, 95% confidence interval [CI] = 1.18-6.37, P < 0.05), APC loss (HR = 8.43, 95% CI = 2.48-28.66, P < 0.01), and BCL2L1 gain (HR = 3.45, 95% CI = 1.13-10.52, P < 0.05) and the independent protective factor was FBXW7 loss (HR = 0.32, 95% CI = 0.12-0.89, P < 0.05). By stepwise Cox regression analysis, three CNAs related to metastasis-free survival were screened out: MDM4 gain (HR = 2.71, 95% CI = 1.11-6.64, P < 0.05), APC loss (HR = 7.19, 95% CI = 1.88-27.60, P < 0.005), and FBXW7 loss (HR = 0.16, 95% CI = 0.05-0.46, P < 0.01). There were significant differences in metastasis-free survival rate between the HCC patients with FBXW7 loss and without MDM4 gain or APC loss, those with MDM4 gain and/or APC loss and without FBXW7 loss, and those with other CNA combinations (log-rank test, P < 0.01). Conclusion: MDM4 gain, APC loss, and FBXW7 loss are the independent prognostic factors for extrohepatic metastasis-free survival after the operation for HCC and can be used to predict the risk of extrohepatic metastasis after the operation for HCC.

Loss at 16q22.1 identified as a risk factor for intrahepatic recurrence in hepatocellular carcinoma and screening of differentially expressed genes.

Copy number alteration (CNA) of chromosome 16, a frequent genetic event in tumors including hepatocellular carcinoma (HCC), has been associated with HCC etiology of hepatitis B virus (HBV) and with clinical outcomes in multiple types of cancer. This study identified CNAs in chromosome 16 in relation to intrahepatic recurrence of HCC in a population with high HBV prevalence, and further screened for differentially expressed genes in recurrence-related CNAs. Array comparative genomic hybridization and expression arrays were used to detect CNAs and gene expression differences, respectively. The associations between CNAs and intrahepatic recurrence were analyzed on 66 patients, follow-up period of 3-73 months. One hundred and nine cases were further evaluated regarding the differentially expressed genes. Losses at 16q and 16p were detected in 62.1% and 51.5% of the 66 cases, respectively. The most recurrent CNAs (with frequency >20%) were losses at 16p13.3-13.2, 16p13.11, 16q11.2-22.1, 16q22.1, 16q22.2-24.2 and 16q24.2. Of the CNAs, 16q22.1 loss was significantly associated with unfavorable intrahepatic recurrence-free survival (P = 0.025). Multivariate Cox analysis identified 16q22.1 loss as an independent risk factor for intrahepatic recurrence (HR = 2.32, 95% CI = 1.26-4.27). A panel of 21 genes, including TRADD, PSMB10, THAP11, CTCF and ESRP2, were significantly downregulated in HCCs with 16q22.1 loss compared to those without the loss. These results suggest that loss at 16q22.1 was associated with increased risk for intrahepatic recurrence of HCC, at least in the HBV-prevalence population. Multiple downregulated genes correlated with the loss were screened.

[Molecular mechanism and pathological evaluation strategies for recurrence and metastasis of liver cancer].

The strong invasiveness and metastasis of hepatocellular carcinoma (HCC) are not only the important pathological mechanisms of high recurrence rate and poor long-term outcome, but also the important pathological basis for developing individualized regimens to control metastasis and recurrence in clinical practice. This article introduces the important molecular events in invasion-metastasis cascade discovered in recent years, including the formation mechanism of metastatic niches, methods for molecular pathological evaluation, and standardized pathological diagnosis.

Overexpression/amplification of HER-2/neu is uncommon in hepatocellular carcinoma.

BACKGROUND: Hepatocellular carcinoma (HCC) is one of the most prevalent fatal cancers in the world. Despite advances in early diagnosis and improvements in surgical techniques, the survival of patients with HCC even after resection is poor because of the high incidence of recurrences. Therefore, the identification of prognostic factors may be helpful in the development of new treatment protocols. AIMS: To investigate HER-2/neu status in HCC by immunohistochemistry (IHC) and fluorescence in situ hybridisation (FISH), and to explore the possibility of using trastuzumab in the treatment of HCC. METHODS: Eight hundred and sixty eight surgical samples from patients with primary HCC were examined for their HER-2/neu status. IHC for HER-2/neu was performed with the HercepTest kit; FISH analysis was performed with the PathVysion HER-2 DNA probe kit. The correlations between HER-2/neu overexpression and clinicopathological characteristics were analysed statistically. RESULTS: HER-2/neu overexpression was detected in 21 (2.42%) of the 868 primary HCCs. Only one specimen showed HER-2/neu gene amplification by FISH. No significant associations were found between HER-2/neu overexpression and the clinicopathological parameters. CONCLUSIONS: There is a low frequency of HER-2/neu overexpression/amplification in HCC. There appears to be no role for HER-2/neu as a prognostic marker and no benefit of anti-HER-2/neu trastuzumab treatment in patients with HCC.

Cloning and characterization of a novel gene (C17orf25) from the deletion region on chromosome 17p13.3 in hepatocelular carcinoma.

Using a combination of hybridization of PAC to a cDNA library and RACE technique, we isolated a novel cDNA, designated as C17orf25 (Chromosome 17 open reading frame 25, previously named it HC71A), from the deletion region on chromosome 17p13.3. The cDNA encodes a protein of 313 amino acids with a calculated molecular mass of 34.8 kDa. C17orf25 is divided into 10 exons and 9 introns, spanning 23 kb of genomic DNA. Northern blot analysis showed that the mRNA expression of C17orf25 was decreased in hepatocellular carcinoma samples as compared to adjacent noncancerous liver tissues from the same patients. The transfection of C17orf25 into the hepatocellular carcinoma cell SMMC7721 and overexpression could inhibit the cell growth. The above results indicate that C17orf25 is a novel human gene, and the cloning and preliminary characterization of C17orf25 is a prerequisite for further functional analysis of this novel gene in human hepatocellular carcinoma.

Multiple genetic alterations involved in the tumorigenesis of human cholangiocarcinoma: a molecular genetic and clinicopathological study.

PURPOSE: Cholangiocarcinoma (CC) is the second most common malignant tumor in the liver and the molecular genetic alterations involved in the tumorigenesis of CC have not been well studied. PATIENTS AND METHODS: The authors analyzed the loss of heterozygosity (LOH) in four tumor suppressor genes, including the adenomatous polyposis coli (APC) gene, the deleted in colon cancer (DCC) gene, the 8-hydroguanine-specific DNA glycosylase (OGG1) gene, and the p53 gene in 22 surgically resected primary CCs by using microdissection-based PCR amplification and direct DNA sequencing. RESULTS: A total of 19 (86.4%) out of 22 CCs exhibited genetic alterations, of which 11 (57.9%) and eight (42.1%) cases showed one and more than one gene alterations, respectively. The frequency of genetic alterations of the four genes studied ranged in order from high to low as APC (68.8%) > DCC (46.2%) > OGG1 (41.7%) > p53 (37.5%). Based on the pattern of altered genes and their correlation with clinical and pathological parameters, the genetic alterations were classified into three groups: group I: no detectable genetic alterations (n = 3, 13.6%); group II: LOH in APC and/or DCC (n = 9, 40.9%); and group III: LOH in OGG1 and/or p53 occurred separately or combined with LOH in APC and/or DCC (n = 10, 45.5%). The > or = 3-year survival rates between group II and group III are 88.9% and 30%, respectively (P < 0.05). No significant differences were found between genetic alterations and tumor size, tumor type, tumor invasion, TNM staging, and tumor differentiation (P > 0.05). CONCLUSION: Accumulation of multiple genetic alterations are involved in the tumorigenesis of CC, of which genetic alterations of APC and DCC occur at a relatively early stage, and of OGG1 and p53 occur at a relatively late stage during development of CC.

Focal nodular hyperplasia with concomitant hepatocellular carcinoma: a case report and clonal analysis.

This report describes a hepatocellular carcinoma (HCC) with concomitant focal nodular hyperplasia (FNH) in a 56 year old Chinese man. There were two well circumscribed tumours measuring 3 x 2.5 x 2 cm and 2 x 1.5 x 1.5 cm. The larger mass was grey and soft with a small area of bleeding and necrosis and an intact capsule. The smaller mass was yellow and had no capsule. Clonal analysis was carried out to clarify the relation between the HCC and the adjacent FNH. The clonal analysis was based on the methylation pattern of the polymorphic X chromosome linked androgen receptor gene (HUMARA). In FNH, after HpaII digestion, the allelic bands showed two well defined peaks. The intensity of the two peaks in the DNA from cirrhotic tissue did not differ significantly, consistent with a random pattern of X chromosome inactivation. However, in HCC, after HpaII digestion, the allelic bands differed significantly in intensity. Therefore, there was a typical polyclonal pattern of inactivation in FNH but the HCC was interpreted as being monoclonal.

Clinicopathological risk factors and prognostic evaluation in hepatocellular carcinoma recurrence after surgery.

AIM: To analyze the clinicopathological risk factors in hepatocellular carcinoma recurrence after surgery. METHODS: We used significance testing (χ(2) and Student's t-test) of single and multiple factors, and Wilcoxon Cox tropic examination; a retrospective clinicopathological analysis was performed on 156 cases of hepatocellular carcinoma after hepatectomy. RESULTS: Of the 156 cases, 68.4%, 57.3%, 46.7%, 31.5%, and 28.6% had one, two, three, four, and five postoperative tumor-free years, respectively; the total recurrence rate was 53.2% (83/156). In the 83 recurrent cases, 65 were intrahepatic subclinical, with a resection rate of 78.3% (65/83). The relevant factors involved in recurrence were: male gender, tumor number and size, capsule infiltration, and portal vein involvement. These factors were an obvious influence on the prognosis of the patients with postoperative hepatocellular carcinoma (P < 0.05). In the recurrent liver carcinomas, 63.1% of tumor nodes (41/65) were at the ipsilateral segment of the primary tumor nodes. CONCLUSION: Male gender, tumor number and size, capsule infiltration, and portal vein involvement are factors for postoperative hepatocellular carcinoma recurrence. Recurrence is mainly unicentral. The right front liver lobe is the segment with a high rate of recurrence.

Small hepatocellular carcinoma. DNA content and biological characteristics.

The DNA content of 12 small hepatocellular carcinomas (SHCC, less than 3 cm in diameter) and 26 large hepatocellular carcinomas (LHCC) was quantitatively determined by means of TV-image analysis. The results showed that 8 patients (66.7%) with SHCC had DNA stem lines in diploid (2C), and 24 (92.3%) with LHCC had DNA stem lines in aneuploid (AN) (P less than 0.01). The incidences of tumor capsule breaking and cancerous thrombosis were 16% and 20% respectively in SHCC, significantly lower than 84% and 80% in LHCC (P less than 0.01). The 5-year survival rate was 75% in patients with SHCC after operation, much higher than 46.2% in patients with LHCC. No relations were found between DNA content, tumor size, pathological grading and serum AFP values of HCC. The results suggest that SHCC less than 3 cm in diameter reflects the early changes of biological characteristics; HCC of 3 cm in size may be at an important period when the changes of DNA stem lines and biological characteristics would occur; SHCC is possible to be determined by serum AFP values; and that the survival rate of the patients can be further improved by early finding and treatment of SHCC less than 3 cm in diameter.

Characteristic changes of DNA stemlines during hepatocarcinogenesis in rats.

DNA stemlines of hepatocellular carcinoma (HCC) model of adult Wistar rats established by diethylnitrosamine were quantitatively measured by flow cytometry. The cancer-inducing course was divided into four stages, eg, precirrhosis stage, cirrhosis stage, early cancer stage and cancer progression stage. The advantageous DNA stemline of hepatocytes of normal adult Wistar rats was tetraploid (4C). It was from the cirrhosis stage that atavistic proliferation of hepatocytes with diploid (2C) DNA stemline started and replaced 4C hepatocytes, resulting in a new advantageous population. The features of early cancer stage were formation of HCC with 2C DNA stemline, whereas during the cancer progression stage, HCC cells with aneuploid (AN) DNA stemline presented the advantageous growth. The study clearly shows the change pattern of DNA stemline during the course of hepatocarcinogenesis from 4C-2C-AN, which is believed to be the biokinetic mechanism of the development and progression of HCC.

[A quantitative study on DNA content of hepatocellular carcinoma by image analysis technique].

DNA content was quantitatively determined in 38 hepatocellular carcinoma (HCC) by image analysis technique using paraffin-embedded sections stained with Feulgen method. The results showed that diploid/near diploid DNA content (D/ND) and aneuploid DNA content (AN) were found in 15.8% and 84.2% of HCC. The incidences of tumor capsule invasion and cancerous thrombus formation were 84.4% and 87.5% in AN HCC which are significantly higher than in D/ND HCC (33.3% and 50%) (P less than 0.05). The tumor size of HCC was related to DNA content, 66.7% of D/ND HCC was less than or equal to 3 cm in diameter, whereas 75% of AN HCC was greater than 3 cm across (P less than 0.05), suggesting that HCC at 3 cm in size may be a critical point at which the biologic characteristics of HCC would change. There was no correlation between the DNA content and pathologic grading of HCC. It is suggested that the development of AN stem line with a broad distribution of DNA content be an important biologic marker of HCC with increased malignancy and the quantitative analysis of DNA content be able to provide an objective and quantitative reference for evaluating the biologic characteristics. The above observation may be taken as a valuable supplement to the pathologic grading of HCC.

[Characteristics and biologic significance of changes in DNA content of rat liver cells during hepatocarcinogenesis].

DNA content of hepatocellular carcinoma (HCC) induced by diethylnitrosamine (DENA) in Wistar rats was quantitatively measured using flow cytometry. In normal adult rats, hepatocytes with tetraploid (4C) DNA content constitute the major cellular population (72.1%), whereas diploid (2C) hepatocytes were of a quite small number (16.9%). Under the persistent effect of DENA, the atavistic proliferation of 2C hepatocytes was obviously increased at the cirrhotic stage, which followed the pre-cirrhotic stage, and replaced 4C hepatocytes to become the major cellular population. HCC at its early cancerous stage was characterized by predominated of DNA 2C cells, and HCC at its progression stage showed a selective and advantageous growth of aneuploid(AN) stemline cells. The whole hepatocarcinogenesis course showed a change of DNA stemline from 4C to 2C to AN, which might be the essential biologic kinetic mechanism of the development and progression of HCC. It is also suggested that HCC at early and progression stages are characterized by predominance of DNA 2c and AN hepatocytes.

[Clinicopathologic features of small hepatocellular carcinoma--an analysis of ninety-three cases].

A clinicopathologic analysis of 93 hepatectomies of < 3 cm small hepatocellular carcinoma (small-HCC) over the last ten years was made, which accounted for 8.5% in 1096 resected HCC during the same period. Serum AFP levels of > or = 400 micrograms/L in patients with small-HCC accounted for 40.3%. The rate of capsule formation was 58.1%, and the incidences of tumor direct invasion, cancer thrombus as well as tumor satellites were 33.3%, 28% and 3.2%, respectively. The 5-year postoperative survival rate of patients with small-HCC was 77.9%. It is proposed that the essential features of < 3 cm small-HCC are: (1) expanding growth pattern and capsule formation in majority of the cases; (2) the lesions are limited, seldom occurring long-distance (> 2 cm) invasion; (3) the incidence of cancer thrombus and satellites are lower; and (4) the majority of < 3 cm small-HCC are of diploid DNA content, showing a relatively slow growth. It is considered that < 3 cm small- HCC basically reflects the pathobiologic features of early HCC, and is an important opportunity of achieving radical therapeutic effect after the resection.