Role of estrogens and epidermal growth factor in hepatocellular carcinoma (HCC)

Estrogen (E) and epidermal growth factors (EGF) receptors were assayed in the liver of nine patients with hepatocellular carcinoma (HCC). Total E and nuclear E receptors were decreased significantly in neoplastic tissue as compared to the levels found in surrounding nonneoplastic tissue. The EGF receptor was decreased also in neoplastic tissue. On the basis of binding data, a decrease in the number but not in affinity of both the E and EGF receptors was found.     

血管内皮生长因子与血小板衍化内皮细胞生长因子在老年人胃癌组织中的表达

目的　探讨老年人胃癌术前活检标本血管内皮生长因子 (VEGF)和血小板衍化内皮细胞生长因子 (PD ECGF)的表达及其与老年胃癌患者预后的关系。　方法　采用免疫组化法检测 92例老年胃癌VEGF、PD ECGF表达情况 ,并分析它们与胃癌临床病理特征关系及对预后的影响。　结果　VEGF、PD ECGF在胃癌组织的表达明显高于慢性萎缩性胃炎 ,进展期癌的表达又高于早期癌(P <0 0 1) ,VEGF、PD ECGF表达呈显著正相关 (相关系数R =0 4 0 5 4 )。VEGF、PD ECGF的阳性表达随着肿瘤大小、浸润深度、TNM分期的递增而呈上调表达 ,有淋巴结转移、血管癌栓的患者表达也明显高于无淋巴结转移、血管癌栓者 (P <0 0 1)。VEGF、PD ECGF阳性表达者总体生存率明显低于VEGF、PD ECGF阴性表达者 (P <0 0 1) ,VEGF、PD ECGF共同表达者生存率更低 (P <0 0 1)。多因素分析表明 ,淋巴结转移、TNM分期、VEGF的表达是老年人胃癌独立的预后因素。　结论　VEGF与PD ECGF表达呈正相关 ,均与胃癌生长、浸润转移关系密切 ,可作为估计老年人胃癌预后的重要因素。     

Platelet-derived growth factor C induces liver fibrosis, steatosis, and hepatocellular carcinoma

Members of the platelet-derived growth factor (PDGF) ligand family are known to play important roles in wound healing and fibrotic disease. We show that both transient and stable expression of PDGF-C results in the development of liver fibrosis consisting of the deposition of collagen in a pericellular and perivenular pattern that resembles human alcoholic and nonalcoholic fatty liver disease. Fibrosis in PDGF-C transgenic mice, as demonstrated by staining and hydroxyproline content, is preceded by activation and proliferation of hepatic stellate cells, as shown by collagen, alpha-smooth muscle actin and glial fibrillary acidic protein staining and between 8 and 12 months of age is followed by the development of liver adenomas and hepatocellular carcinomas. The hepatic expression of a number of known profibrotic genes, including type beta1 TGF, PDGF receptors alpha and beta, and tissue inhibitors of matrix metalloproteinases-1 and -2, increased by 4 weeks of age. Increased PDGF receptor alpha and beta protein levels were associated with activation of extracellular regulated kinase-1 and -2 and protein kinase B. At 9 months of age, PDGF-C transgenic mice had enlarged livers associated with increased fibrosis, steatosis, cell dysplasia, and hepatocellular carcinomas. These studies indicate that hepatic expression of PDGF-C induces a number of profibrotic pathways, suggesting that this growth factor may act as an initiator of fibrosis. Moreover, PDGF-C transgenic mice represent a unique model for the study of hepatic fibrosis progressing to tumorigenesis.     

Circulating vascular endothelial growth factor (VEGF) is a possible tumor marker for metastasis in human hepatocellular carcinoma

Vascular endothelial growth factor (VEGF) is closely related to angiogenesis in various human cancers. However, little is known of its circulating levels in hepatocellular carcinoma (HCC). We examined circulating VEGF levels in chronic liver disease to assess their clinical significance. Plasma VEGF concentrations were determined, by enzyme immunoassay, in patients with chronic hepatitis (CH; n = 36), liver cirrhosis (LC; n = 77), and HCC (n = 86) for a cross-sectional study. Plasma VEGF levels in healthy controls (n = 20) and CH, LC, and HCC patients were 17.7 ± 5.4 (mean ± SD), 30.6 ± 22.8, 34.4 ± 27.0, and 51.1 ± 71.9 pg/ml, respectively. The levels were significantly elevated in the HCC group, compared with the control, CH, and LC groups. Plasma VEGF levels in stage I, II, III, IVA, and IVB HCC patients were 27.6 ± 16.1, 26.5 ± 13.7, 35.8 ± 15.3, 45.4 ± 39.4, and 103.1 ± 123.2 pg/ml, respectively. The stage IVB patients with remote metastasis showed significantly marked elevation compared with the patients at the other stages. Platelet numbers were weakly correlated with plasma VEGF levels in the HCC group. Plasma VEGF level was highly elevated in patients with HCC, particularly those with metastatic disease. We consider that plasma VEGF is a possible tumor marker for metastasis of HCC. Circulating VEGF may be derived mainly from the large burden of tumor cells, and partly from platelets activated by the vascular invasion of HCC cells. (Received June 30, 1997; accepted Oct. 30, 1997)     

Laparoscopic radiofrequency of hepatocellular carcinoma (HCC) in liver cirrhosis

BACKGROUND/AIMS: In this report, the feasibility and efficacy of laparoscopic radiofrequency interstitial thermal ablation of hepatocellular carcinoma has been evaluated in 20 patients. METHODOLOGY: The laparoscopic approach with the use of intraoperative ultrasonography allowed us to obtain additional information regarding liver nodules and a complete treatment of the liver lesions. RESULTS: The complication rate was low and there was no operative mortality. A complete necrosis has been obtained in 90% of the cases at 1 month dynamic computed tomography following the treatment. CONCLUSIONS: Laparoscopic radiofrequency thermal ablation of hepatocellular carcinoma proved to be a safe and effective technique; its use may be proposed in selected patients. Larger series are needed to accurately assess its role among the other ablative therapies of hepatocellular carcinoma.     

Connective tissue growth factor is overexpressed in human hepatocellular carcinoma and promotes cell invasion and growth

AIM: To determine the expression characteristics of connective tissue growth factor (CTGF/CCN2) in human hepatocellular carcinoma (HCC) in histology and to elucidate the roles of CCN2 on hepatoma cell cycle progression and metastasis in vitro.METHODS: Liver samples from 36 patients (who underwent hepatic resection for the first HCC between 2006 and 2011) and 6 normal individuals were examined for transforming growth factor β1 (TGF-β1) or CCN2 mRNA by in situ hybridization. Computer image analysis was performed to measure integrated optimal density of CCN2 mRNA-positive cells in carcinoma foci and the surrounding stroma. Fibroblast-specific protein-1 (FSP-1) and E-cadherin were examined to evaluate the process of epithelial to mesenchymal transition, α-smooth muscle actin and FSP-1 were detected to identify hepatic stellate cells, and CD34 was measured to evaluate the extent of vascularization in liver tissues by immunohistochemical staining. CCN2 was assessed for its stimulation of HepG2 cell migration and invasion using commercial kits while flow cytometry was used to determine CCN2 effects on HepG2 cell-cycle.RESULTS: In situ hybridization analysis showed that TGF-β1 mRNA was mainly detected in connective tissues and vasculature around carcinoma foci. In comparison to normal controls, CCN2 mRNA was enhanced 1.9-fold in carcinoma foci (12.36 ± 6.08 vs 6.42 ± 2.35) or 9.4-fold in the surrounding stroma (60.27 ± 28.71 vs 6.42 ± 2.35), with concomitant expression of CCN2 and TGF-β1 mRNA in those areas. Epithelial-mesenchymal transition phenotype related with CCN2 was detected in 12/36 (33.3%) of HCC liver samples at the edges between carcinoma foci and vasculature. Incubation of HepG2 cells with CCN2 (100 ng/mL) resulted in more of the cells transitioning into S phase (23.85 ± 2.35 vs 10.94 ± 0.23), and induced a significant migratory (4.0-fold) and invasive (5.7-fold) effect. TGF-β1-induced cell invasion was abrogated by a neutralizing CCN2 antibody showing that CCN2 is a downstream mediator of TGF-β1-induced hepatoma cell invasion.CONCLUSION: These data support a role for CCN2 in the growth and metastasis of HCC and highlight CCN2 as a potential novel therapeutic target.     

Platelet-derived endothelial cell growth factor: structure and function.

Platelet-derived endothelial cell growth factor (PD-ECGF) is a 45-kDa single chain polypeptide, which stimulates the growth and chemotaxis of endothelial cells in vitro and angiogenesis in vivo. Purification from human platelets and cDNA cloning of PD-ECGF disclosed that it is a novel type of angiogenic factor without sequence similarity to hitherto known proteins. PD-ECGF is present in human platelets as well as in placenta. Amino acid sequencing of PD-ECGF from human placenta revealed that the placental form has an additional 5 amino acids at the N-terminus. In cultured cells, it is produced by normal fibroblasts as well as some transformed cell lines. PD-ECGF lacks a hydrophobic signal sequence and remains inside the producer cells. PD-ECGF may act at sites of injury as a wound hormone and thus play an important role under several physiological and pathological conditions.     

Platelet-derived growth factor (PDGF) receptor activation in cell transformation and human malignancy

We demonstrate that the v-sis-transformed NIH/3T3 fibroblasts exhibit tyrosine-phosphorylation of both intracellular and cell surface forms of the and β platelet-derived growth factor (PDGF) receptors (PDGFRs). Cell proliferation was partially inhibited by PDGF-neutralizing antibody, but was completely blocked by the drug suramin. Suramin treatment markedly reduced tyrosine-phosphorylated cell surface PDGFRs, but had no effect on the tyrosine-phosphorylated intracellular receptor species. These findings indicate that v-sis-activated PDGFRs must attain a cell surface localization to functionally couple with the mitogenic-signaling pathway. Additionally, we were able to demonstrate a functional autocrine loop involving PDGF in human tumor cell lines. Exposure to suramin resulted in diminished receptor autophosphorylation and/or upregulation of the PDGFRs. A subset of the tumor cell lines possessing a PDGF autocrine pathway exhibited a significant reduction in proliferation after exposure to suramin. These findings indicate that a PDGF autocrine loop contributes to the uncontrolled proliferative drive in some human malignancies.     

Expression of platelet-derived endothelial cell growth factor and vascular endothelial growth factor in hepatocellular carcinoma and portal vein tumor thrombus

Purpose: Both platelet-derived endothelial cell growth factor (PD-ECGF) and vascular endothelial growth factor (VEGF) are known to promote the development of new blood vessels, which are fundamental to tumor growth and metastasis. We aimed at evaluating the gene expression of PD-ECGF and VEGF in hepatocellular carcinoma (HCC) and portal vein tumor thrombus (PVTT). Patients and methods: Surgical specimens (28 HCC, 28 nontumorous liver tissues and 18 PVTT) were studied by Northern blot analysis. The levels of PD-ECGF mRNA and VEGF mRNA expression were measured by densitometric scanning of the autoradiographs, and they were normalized to the level of expression of an internal control (glyceraldehyde-phosphate dehydrogenase) mRNA. Results: The expression rates of PD-ECGF mRNA in PVTT, HCC and nontumorous liver tissues were 77.8% (14/18), 67.9% (19/28) and 35.7% (10/28), being 88.9% (16/18), 75.0% (21/28) and 17.9% (5/28) respectively for VEGF mRNA. The expressions of PD-ECGF mRNA and VEGF mRNA were higher in HCC with PVTT than when PVTT was absent (P < 0.05). The PVTT was more often seen in patients with positive expression of both PD-ECGF mRNA and VEGF mRNA in HCC than in patients who were positive for only one of these factors or negative for both (P < 0.05). Conclusion: Both PD-ECGF and VEGF correlated well with the formation of PVTT of HCC. Received: 20 June 1999 / Accepted: 20 July 1999     

Laparoscopic liver resections for hepatocellular carcinoma (HCC) in cirrhotic patients

BackgroundThe laparoscopic approach for liver resections is still limited and controversial. Nevertheless the advantages connected with a mini-invasive approach are significant, especially in cirrhotic patients. In recent years the progress of laparoscopic procedures and the development of new and dedicated technologies have made endoscopic hepatic surgery feasible and safe. The aim of this study was to report the results of our experience in laparoscopic liver surgery for hepatocellular carcinoma (HCC) in cirrhotic patients.MethodsFrom 2000 to 2003, 16 patients (10 male, 6 female; age 48–69 years; mean age 60.1 years) with HCC and associated severe but well compensated liver cirrhosis underwent laparoscopic hepatic resections at our department. Mean tumour size was 2.9 cm (range 1–3.9). Seven of these lesions were in the left liver and nine in the right lobe. Laparoscopy was performed under CO₂ pneumoperitoneum. The liver was always examined using laparoscopic ultrasound (US) to confirm the extension of the lesions and their relationships to the vasculature. The Pringle manoeuvre was not used. The transection of liver parenchyma was obtained by the use of a harmonic scalpel. The specimens were placed in a plastic bag and removed without contact to the abdominal wall.ResultsThere was one conversion to laparotomy for inadequate exposure. In the remaining 15 patients we performed 13 non-anatomical resections, I segmentectomy and I anatomical left lobectomy. The mean operative time was 152 min (range 80–180). Mean blood loss was 280 ml and none of the patients required blood transfusions. In two patients the resection margin was <1 cm but the capsule was not infiltrated at histology. One patient died on the third postoperative day from a severe respiratory distress syndrome. Major morbidities occurred in two patients who developed moderate postoperative ascites, which resolved successfully with conservative treatment. The mean postoperative hospital stay was 8.8 days. Mean follow-up time has been 18 months, and to date no recurrences at the site of resection or port-site metastases have been observed.DiscussionLimited laparoscopic liver resections in cirrhotic patients are technically feasible with a low complication rate when careful selection criteria are followed (hepatic involvement limited and located in the left or anterior right segments, tumour size smaller than 5 cm, Child-Pugh class A). This approach could be considered the best option for the treatment of small esophitic or subcapsular HCC on well compensated cirrhosis and a useful option when it is necessary to perform a left lateral anatomical resection or non-anatomical resection in well selected patients. In fact the mini-invasive approach can minimise the postoperative morbidity rate, which is still too high in this group of patients. It must be performed in highly specialised units by surgeons assisted by all requested technologies and with extensive experience in hepatobiliary and advanced laparoscopic surgery.     

肝癌组织血管内皮生长因子表达水平的免疫组化研究

目的　旨在研究血管内皮生长因子 (VEGF)与肝癌微血管形成、生长和转移诸方面的关系。方法　对临床 3 6例肝癌术后癌组织 ,以免疫组织化学法研究VEGF在肿瘤组织的胞内分布及其表达 ;并以ELISA法测定癌灶、癌旁及远癌组织中的VEGF蛋白的表达水平。结果　癌组织中VEGF阳性表达率为 63 .9% ;无包膜或包膜不完整组VEGF阳性表达率与有包膜组存在显著差异 ;肝癌伴有远处转移组VEGF阳性表达水平显著高于无转移组 (P <0 .0 1) ,癌灶组织中VEGF的表达水平明显高于癌旁、远癌组织(P <0 .0 1)。结论资料提示VEGF在肝癌组织中高度表达 ,它在HCC的血管形成、肿瘤发展和转移过程中起重要作用 ,提示癌组织中VEGF过度表达是反映肿瘤侵袭生长及转移潜能的有效指标     

膀胱移行细胞癌VEGF和VEGFR的表达及相关性的探讨

目的　探讨膀胱移行细胞癌 (BTCC)中血管内皮生长因子 (VEGF)及其受体 (VEGFR)的表达及与两者之间的关系。方法　采用免疫组织化学链霉菌抗生物素 过氧化物酶连接法 (S P法 )对 30例BTCC及 1 0例正常膀胱黏膜组织中VEGF及VEGFR的表达进行检测。结果　VEGF和VEGFR在绝大多数BTCC中呈阳性表达 ,平均表达率分别为 87%和 73 %。随肿瘤分期和分级的升高其表达水平升高 ,但在正常膀胱组织中未见表达。结论　BTCC中VEGF和VEGFR表达阳性 ,提示其在BTCC的血管生成和侵袭进展过程中起着重要作用 ,并将有可能为BTCC抗血管形成治疗及预防提供新的思路     

Characteristics of a cell line (Tong/HCC) established from a human hepatocellular carcinoma

A continuous adherent cell line was established from a hepatocellular carcinoma of an HBsAg-positive Italian male. This cell line, designated Tong/HCC, has been grown in a hormone-supplemented medium for more than 18 months. The cell line secretes HBsAg, alpha-fetoprotein, albumin and alpha 1-antitrypsin. alpha-Fetoprotein production is enhanced by the addition of hydrocortisone and appears to be glucocorticoid concentration-dependent. The concentrates of the supernatant from the cell cultures and cell lysates were negative when tested for HBeAg. The cell culture medium was negative for hepatitis B virus DNA when tested by dot-blot hybridization. However, hepatitis B virus DNA was found to be integrated in the chromosomal DNA by Southern blot analysis. At least five different integration sites were identified, and no free hepatitis B virus DNA was observed. The modal chromosome number was 64, and a translocation on Chromosome 15 was consistently noted. HLA typing revealed sites for A3, Aw24, Bw34 and Cw1.     

Pathobiology of chronic hepatitis virus infection and hepatocellular carcinoma (HCC).

Hepatitis viruses may cause liver cancer (HCC) through an indirect mechanism inducing inflammation and cirrhosis. Only hepatitis B virus (HBV) was shown to have a direct oncogenetic potential. Hepatitis D virus (HDV) infection, superimposed on the oncogenetic background provided by chronic HBV infection, appears to provide an additional risk for HCC. Patients with florid infections from both HBV and HDV and active liver inflammation develop HCC at a significantly younger age than those infected by HBV alone or infected by hepatitis C virus (about 10 years earlier). In patients positive for serum HBV-DNA/HDV-RNA and/or IgM anti-HBc/IgM anti-HD it is mandatory to program a more frequent (thrice a year) schedule of screenings (ultrasound scan, alpha-1-phetoprotein, etc.) for prophylaxis of HCC.     

High serum levels of vascular endothelial growth factor in patients with human hepatocellular carcinoma

Vascular endothelial growth factor (VEGF) is intimately involved in neovascularization. In addition, it is know that in human hepatocellular carcinoma (HCC), angiogenesis is indispensable for tumor growth. In this study, we measured the serum VEGF levels of patients with HCC and studies the relationship between the serum VEGF level and maximum tumor diameter as well as that between the serum VEGF level and the serum α-fetoprotein (AFP) level. Mean serum VEGF level were 5.33 ± 0.77, 3.97 ± 0.68, 2.64 ± 0.78, and 2.57 ± 0.97 ng/ml for patients with HCC, chronic hepatitis (CH), or liver cirrhosis (LC) and normal controls (NC), respectively, with that of the HCC patients being significantly (P < 0.05) higher than that of the LC patient or NC. In addition, the serum VEGF level was significantly (r = 0.53, P < 0.05) correlated with the maximum tumor diameter in the HCC patients, and the sera of the patients with hypervascular HCC showed a significantly (P < 0.01) higher VEGF titer than the sera of the patients with isovascular or hypovascular HCC. However, there was no significant correlation between serum VEGF level and serum AFP level. These findings suggest that VEGF may play an important role, apart from that in AFP production, in the development of HCC.