The expression levels of transcription factors T-bet, GATA-3, RORγt and FOXP3 in peripheral blood lymphocyte (PBL) of patients with liver cancer and their significance.

OBJECTIVES: To investigate the expression of transcriptional factors (TFs) T-bet, GATA-3, RORγt and FOXP in peripheral blood mononuclear cells (PBMC) of patients with hepatocellular carcinoma (HCC) and to evaluate the correlation between the imbalances of Th1/Th2, Th17/Treg at the expression levels and liver cancer Methods: The peripheral venous blood was drawn from 20 HCC-patients (HCC-group) and 20 health participants (C-group). The expression levels of Th1, Th2 and Th17 and the major Treg-specific TFs T-bet, GATA-3, RORγt and FOXP3 in the PBMC were measured with quantitative real-time PCR(RT-qPCR). RESULTS: The mRNA level of Th1-specific TF T-bet in HCC-group was significantly lower than that of C-group (52.34±34.07 VS 104.01±56.00, P<0.01); the mRNA level of Th2-specifc TF, GATA-3, in HCC group was significantly higher than that in C-group (1.38±1.15 VS 0.58±0.65, P<0.05) and T-bet mRNA/GATA-3 mRNA ratio was significantly lower in HCC-group than in C-group (86.01±116.71 VS 461.88±708.81, P<0.05). The mRNA level of Th17-specific TF RORγt in HCC-group was significantly higher than that of C-group (72.32±32.82 VS 33.07±22.86, P<0.01). Treg-specific TF FOXP3 mRNA level was significant higher in HCC-group than in C-group (3.17±1.59 VS 1.39±1.13, P<0.01) CONCLUSION: T-bet mRNA level was reduced whereas GATA-3 mRNA level was increased and T-bet/GATA-3 ratio was significantly reduced in PBMC, indicating that Th1/Th2 ratio was of imbalance at TF levels in PBMC of HCC, displaying Th2 thrift phenomena. The mRNA levels of RORγt and FOXP3 in PBMC of HCC were significantly increased, indicating the existence of a predominant phenomenon of Th17- and Treg-expressing PBMC in HCC.

Maturation induction of human peripheral blood mononuclear cell-derived dendritic cells.

The aim of the present study was to explore an optimal method for maturation induction of dendritic cells (DCs). Human monocyte-derived DCs were induced in the presence of GM-CSF and IL-4. On Day 6, the maturation of DCs was induced with CD40L, LPS, TNF-α and cocktail of cytokines (TNF-α, IL-6, IL-1ß and PGE2), respectively, for 24 h. Then, DCs were harvested and subjected to flow cytometry (FCM) for the detection of CD80, CD83, CD86 and HLA-DR. FITC-dextran endocytic activity was measured by FCM, IL-12 production by ELISA and T lymphocyte proliferation following DC stimulation by MTT assay. CD40L, LPS, TNF-α and a cocktail of cytokines induced DC maturation. Induction with the cocktail of cytokines was the most efficient, and the expression rate of CD83 was 66.91% (P<0.05). The FITC-dextran endocytic activity of mature DCs was significantly reduced, and IL-12 production was dramatically increased in mature DCs, particularly in those following induction using the cocktail of cytokines. The mature DCs had potent ability to stimulate the proliferation of lymphocytes. The cocktail of cytokines is a favorable strategy for the induction of DC maturation.

Staging systems for predicting survival of patients with hepatocellular carcinoma after surgery.

AIM: To compare the staging systems for stratifying and predicting the prognosis of patients with hepatocellular carcinoma (HCC) after partial hepatectomy (PH). METHODS: Clinical data about 438 HCC patients who underwent PH from January 1991 to December 2004 at our hospital were retrospectively analyzed. Tumor stage was evaluated following the Chinese tumor node metastasis (TNM) and barcelona clinic liver cancer (BCLC) staging systems, respectively. Survival curves for the HCC patients were plotted using the Kaplan-Meier method and differences were compared by the log-rank test. The accuracy of each system for predicting death of HCC patients was evaluated by calculating the area under the receiver operating characteristic curve. RESULTS: The HCC patients were classified into stages I-III, stages I-IV and stages A-C, according to the 3 staging systems, respectively. Log-rank test showed that the cumulative survival rate was significantly different for the HCC patients at 3 Chinese system stages, TNM stages I and II, TNM stages III and IV, and 3 BCLC stages (P < 0.05). However, no significant difference was found in the HCC patients at TNM stages II and III. The accuracy of the Chinese and BCLC staging systems was higher than that of the TNM staging system for predicting the survival rate of HCC patients. CONCLUSION: The Chinese and BCLC staging systems are better for stratifying and predicting the prognosis of HCC patients after PH than the TNM staging system.

Prognostic factors and outcome of 438 Chinese patients with hepatocellular carcinoma underwent partial hepatectomy in a single center.

OBJECTIVE: This study was designed to evaluate the prognostic factors and outcome of 438 Chinese patients with hepatocellular carcinoma who underwent partial hepatectomy in a single center. METHODS: Clinicopathological data of 438 patients with hepatocellular carcinoma who underwent partial hepatectomy at the author's hospital between 1991 and 2004 were reviewed retrospectively. The Kaplan-Meier method was adopted for evaluating survival. Prognostic factors were assessed by Cox proportional hazard model and logistic regression model. RESULTS: The perioperative (30 days) mortality and morbidity were 7.5% (33/438) and 21.7% (95/438), respectively. The operative mortality decreased significantly from 10.6% (23/218) in 1991-2001 to 4.5% (10/220) in 2002-2004 (P = 0.019). Postoperative overall survival rates at 1 year, 3 years, and 5 years were 72.2%, 53.5%, and 43.3%, respectively. Cox multivariate analysis indicated that Child-Pugh score, tumor size, capsular invasion, tumor stage, vascular invasion, and resection margin were independent prognostic factors for overall survival (P < 0.05). Also, 254 cases had tumor recurrence after operation and 87 cases of them were reoperated. Logistic multivariate analysis showed that tumor size, capsular invasion, vascular invasion, lymph node metastasis, extrahepatic metastasis, and resection margin were independent risk factors of tumor recurrence (P < 0.05). CONCLUSIONS: Tumor size, capsular invasion, vascular invasion, and resection margin were the main factors that may impact the overall survival and tumor recurrence. Because resection margin was the only factor that relates to the surgery, enough resection margin (>2 cm) should be obtained whenever possible.

[The effects of recombinant human growth hormone on the proliferation of Bel-7402 human hepatic carcinoma cell lines in vitro and its regulation on growth hormone receptors].

OBJECTIVE: To investigate the role of recombinant human growth hormone (rhGH) in the growth of Bel-7402 human hepatic carcinoma cell line (Bel-7402 line) in vitro and its effects on GHR expression. METHODS: Tumor cell count, MT assay and colony forming test were performed to determine the responses of Bel-7402 to different concentrations of rhGH (0, 1, 10, 100, 1000, 10 000 ng/ml). Metabolism of DNA in tumor cells was analyzed with the method of mixture of 3H-TdR. Radioreceptor assay was used to detect the GHR expression of the hepatic carcinoma cell lines and its relation to different rhGH concentrations. RESULTS: rhGH accelerated the proliferation of the Bel-7402 line when the concentration of rhGH was over 100 ng/ml (P < 0.05). Other rhGH concentrations had also positive effects, but with reduced effect as compared with that of 100 ng/ml. After 24 h of rhGH addition of concentration of 10 ng/ml and 100 ng/ml, GHR site number was significantly higher than that in control group, while the 10,000 ng/ml group showed a significantly lower GHR site number. CONCLUSIONS: Different concentrations of rhGH might result in variable effects on the growth of Bel-7402 hepatic carcinoma cell line. Certain concentrations of rhGH might stimulate the growth of the cell line. rhGH can regulate the expression of GHR in the cell line.

Vascular damage and anti-angiogenic effects of tumor vessel-targeted adenovirus-mediated herpes simplex virus thymidine kinase gene.

AIM: To explore the therapeutic efficacy and mechanism of herpes simplex virus-thymidine kinase (HSV-tk) targeting angiogenesis against hepatocellular carcinoma in vivio and in vitro. METHODS: Recombinant adenovirus containing kinase domain insert with receptor (KDR) or cytomegalovirus (CMV) promoter-controlled HSV-tk gene (AdKDR-tk and AdCMV-tk) was constructed using pAdeasy system. The expression of KDR antigen in human umbilical venous endothelial cells (HUVEC) and HepG2 was detected with histological analysis of cells. The virus was used to infect HUVEC and HepG2. Following administration of ganciclovir (GCV), the survival rate of gene-transfected HUVEC and HepG2 was evaluated by MTT method. To develop hepatocarcinomas in 32 Balb/C mice with HepG2 cells, the mice were divided into four groups: ganciclovir group (I), Ad group (II), AdCMV-tk group (III) and AdKDR-tk group (IV). Then selective administration of recombinant adenovirus or Ad via the intratumorial was given to all rats. Ganciclovir (GCV) was given at a dose of 100 mg kg(-1) d(-1) (ip) started on the following day and lasted 10 d. Microvessel density (MVD) of tumor in all the treated animals were examined by the immunohistochemical methods and tumor burden was evaluated 10 d before and after the last GCV dose. RESULTS: Immunocytochemical staining indicated the expression of KDR antigen in HUVEC. Under adenovirus infection index of 100, with increasing GCV concentration from 0 up to 50 mg/L, the survival rate of AdKDR-tk-transfected HUVEC and HepG2 decreased from 100% to (28.94 +/- 5.67)% and (75.45 +/- 2.91)% at proper order, respectively (P < 0.01), while the survival rate of AdCMV-tk-transfected HUVEC and HepG2 declined from 100% to (17.56 +/- 2.48)% and (23.15 +/- 5.72)%, respectively (P > 0.05). Compared with group I, there was a decrease of tumor weight by 14.7% in group III and by 23.6% in group IV. And there was a distinct difference between group III and IV (P < 0.05). The median MVD for all groups was 37.4 +/- 8.6, 30.6 +/- 7.8, 27.6 +/- 7.1, and 10.7 +/- 4.1 (microvessels/mm(2)) in group I, II, III and IV, respectively. And there was a marked difference between group III and II (P < 0.05), IV and II (P < 0.01), and IV and III (P < 0.01). CONCLUSION: KDR promoter-HSV-tk gene may effectually restrain the growth of tumor via targeting angiogenesis for hepatocellular carcinoma with treatment of GCV.

A 26-year clinical observation of splenic auto-transplantation and oesophageal transection anastomosis: a new treatment strategy in patients with portal hypertension.

BACKGROUND: Surgical treatment options for patients with cirrhosis and portal hypertension are complicated. In this study, we evaluated the effectiveness of a new treatment strategy, splenic auto-transplantation and oesophageal transection anastomosis. We report results from clinical observations, splenic immune function and portal dynamics in 274 patients. METHODS: From 1979 to 2005, 274 cirrhosis patients with portal hypertension underwent the new treatment strategy, and were followed up to compare results with those patients who underwent traditional surgical treatment. From 1999 to 2002, a randomized controlled trial (RCT) was performed on 40 patients to compare their post-operative immune function. From 1994 to 2006, another RCT enrolled 28 patients to compare portal dynamics using three-dimensional dynamic contrast-enhanced magnetic resonance angiography (3D DEC MRA) investigation post operation. RESULTS: Among 274 patients (mean age 41.8 years), the emergency operative mortality (4.4%), selective operative mortality (2.2%), complication rate (17.9%), prevalence of hepatic encephalopathy (< 1%), rate of portal hypertension gastritis (PHG) bleeding (9.1%), and morbidity of hepatic carcinoma (8%) were similar to those patients undergoing traditional operation; the spleen immunology function (Tuftsin, IgM) decreased in both groups 2 months post operation, but this decrease did not reach statistical significance. Through 3D DCE MRA, the cross sectional area and the velocity and volume of blood flow of the main portal vein decreased significantly after operation in both groups. The velocity and volume of blood flow in the auto-transplantation group was significantly lower than that in the control group. CONCLUSIONS: Splenic auto-transplantation and esophageal transection anastomosis is a safe, effective, and reasonable treatment strategy for patients with portal hypertension with varicial bleeding. It not only can correct hypersplenism, but may also achieve complete hemostasis. Spleens auto-transplanted into the retroperitoneal space can preserve immune function and establish broad collateral circulation.

[Effect of recombinant human growth hormone on growth of human Bel-7402 hepatic carcinoma xenografts in nude mice].

BACKGROUND & OBJECTIVE: Recombinant human growth hormone (rhGH) has been widely used in clinical medicine; while for the patients with hepatocellular carcinoma (HCC), if rhGH could cause tumor growth, metastasis or recurrence other than improve patients' body condition is uncertain. Because most primary hepatic carcinoma could express human growth hormone receptor (GHR), this study was to explore the effects of rhGH on the growth of human Bel-7402 hepatic carcinoma (with GHR expression) xenograft in nude mice. METHODS: The expression of GHR was confirmed in Bel-7402 cells by radioligand assay. After transplantation of Bel-7402 hepatic carcinoma in liver, 32 nude mice were randomly divided into 4 groups. Except 1 control group, 3 experimental groups were injected with different dosages of rhGH [0.5, 1.0, and 2.0 U . (kg.day)(-1), respectively] subcutaneously 2 weeks after tumor transplantation, once a day for 2 weeks. All nude mice were killed at the end of the experiment to observe the growth of xenografts. The expression of proliferating cell nuclear antigen (PCNA) in xenografts was examined by immunochemistry. RESULTS: The tumor weight was significantly heavier and the tumor volume was significantly larger in 1.0 U . (kg.day)(-1) rhGH and 2.0 U . (kg.day)(-1) rhGH groups than in control group (P<0.05), but no significant difference was found between 0.5 U . (kg.day)(-1) rhGH group and control group (P>0.05). Labeling index (LI) of PCNA was significantly higher in each experimental group than in control group (P<0.05), and significant differences were also found among the 3 experimental groups (P<0.05). The dosage of rhGH was positively correlated to LI of PCNA in tumor tissues (r=0.779, P<0.001). CONCLUSIONS: rhGH could enhance the growth of Bel-7402 hepatic carcinoma xenografts in nude mice. Its effect positively correlates with rhGH dosage.

[Application of adenosine triphosphate tumor chemosensitive assay system to individual chemotherapy for hepatocellular carcinoma].

BACKGROUND & OBJECTIVE: Present chemotherapy for hepatocellular carcinoma (HCC) is not effective. To improve the effect of chemotherapy, in vitro chemo-drug sensitive testing system, adenosine triphosphate tumor chemosensitive assay (ATP-TCA) system, was used to evaluate efficacies of chemotherapeutic drugs, and to guide clinical individual chemotherapy for HCC patients. METHODS: ATP-TCA system was applied to test efficacies of 5-fluorouracil (5-FU), mitomycin (MMC), cisplatin (DDP), oxaliplatin (OXA), epirubicin (EPI), gemcitabine (GEM), irinotecan (CPT-11), etoposide (VP-16), and paclitaxel (PTX) on 50 HCC samples. Twenty-three HCC patients received ATP-TCA-directed chemotherapy (ATP-TCA group)û 20 HCC patients received surgery and routine treatments (control group). Clinical outcomes of these patients were observed for 162 weeks. RESULTS: The assessable rate of ATP-TCA result is 90.8%. In the 50 samples, the sensitive (moderate to high degree) rates were 46% to PTX, 44% to CPT-11, 36% to GEM, 14% to MMC, 12% to EPI, 8% to DDP, 6% to VP-16, 6% to OXA, and 4% to 5-FU, respectively. In clinical trial, at the research end-point, no significant differences were found in partial remission (PR), complete remission (CR), stable disease (SD), and mortality between ATP-TCA group and control group (P > 0.05), but progression disease (PD) rate was significantly higher in control group than in ATP-TCA group (60.00% vs.13.04%, P=0.003); significant differences were found in overall response rate (ORR) (60.86% vs. 30.00%, P =0.043), overall survival (OS) (78.91 weeks vs. 27.21 weeks, P=0.006), and progress-free survival (PFS) (30.52 weeks vs. 4.78 weeks, P=0.005) between ATP-TCA group and control group. CONCLUSIONS: ATP-TCA system might be useful in evaluating the efficacy of chemotherapeutic drugs on HCC samples, and in planning individualized chemotherapy regimen for HCC patients. PTX, CPT-11 and GEM might be potential drugs for the treatment of HCC. ATP-TCA-guided chemotherapy might prolong survival time of HCC patients.

Effects of deep low temperature on secondary branches enclosed in Glisson's system of pig hepatic lobes.

BACKGROUND: Hepatic cryosurgery, a safe and effective approach for an unresectable hepatoma, has been extensively applied in clinical setting. But whether direct deep freezing to the tumor involving special location, i.e., Glisson's system of certain hepatic lobe could cause the impairment of the duct system or post impairment effect remains a mystery. Consequently, the aim of the study was to investigate the effect of freezing on the secondary branches enclosed in the Glisson's system of certain hepatic lobes. METHODS: Twenty pigs were divided into 2 groups randomly. The treated group: the area of secondary branches of the Glisson's system in the left lateral lobe was frozen by a cryoprobe at deep low temperature for 3 minutes, with the blockage of blood flow from the porta hepatis. The control group: only the blockage of the porta hepatis was performed for 3 minutes. Serologic examinations, color Doppler examination and pathological observation were used for evaluation of this procedure postoperatively. RESULTS: The frozen hepatic parenchyma, the wall of the frozen secondary bile duct and portal vein showed necrosis. However, the frozen hepatic artery ramification did not show any obvious changes postoperatively. Eight weeks after cryosurgery, the lumen of the hepatic artery and portal vein maintained unobstructed. Meanwhile, atrophy and fibroplasia occurred in the related hepatic lobe. All the animals recovered well. CONCLUSIONS: Animals could tolerate the direct deep-freezing of the area of secondary branches enclosed in the Glisson' s system. The therapeutic effect could be attained by the necrosis of the frozen hepatic parenchyma.

Possible stem cell origin of human cholangiocarcinoma.

AIM: To investigate the expression of CD34 and c-kit (receptor of stem cell factor) in cholangiocarcinoma. METHODS: Fifteen cases of intrahepatic cholangiocarcinoma and 17 cases of extrahepatic cholangiocarcinoma were studied in this experiment. Using Envision detection system, paraffin-embedded sections of the resected cholangiocarcinoma tissue were stained with antibodies against CD34 and c-kit, respectively. The sections were counterstained with hematoxylin, and the results were examined under light microscope. Normal tonsil and mammary tissues were used as positive controls for CD34 and c-kit, respectively. RESULTS: CD34 was positive in all sections, but only in capillary endothelial cells of tumor tissue. No cholangiocarcinoma cells were positive for CD34. In one case of extrahepatic cholangiocarcinoma, a few tumor cells (about 5%) were immunoreactive with c-kit. CONCLUSION: CD34 or c-kit positive cells in liver tissue may represent liver stem cells, as they can differentiate into mature biliary cells in vitro. The expression of c-kit by some cholangiocarcinoma cells suggests that cholangiocarcinoma might originate from liver stem cells. However, other mechanisms of hepatocarcinogenesis, such as de-differentiation of mature cholangiocytes, may also exist.

Protective effects of recombinant human growth hormone on cirrhotic rats.

AIM: To investigate the effects and molecular mechanisms of recombinant human growth hormone (rhGH) on protecting liver function and alleviating portal hypertension of liver cirrhotic rats. METHODS: Liver cirrhosis of male Sprague-Dawley rats was induced by administration of thioacetamide. The rats with or without liver cirrhosis were randomly divided into four groups. Group A consisted of the normal rats was treated with normal saline (NS), group B consisted of the normal rats was treated with rhGH, group C consisted of cirrhotic rats was treated with NS, and group D consisted of cirrhotic rats was treated with rhGH. The rats of different groups were subcutaneously injected with 0.5 mL of NS or 333 ng/kg of rhGH daily for 7 d. After treatments, the following parameters were examined, including GH-binding capacity (R(T)) by (125)I-hGH binding, growth hormone receptor mRNA(GHR mRNA) expression by RT-PCR, relative content of collagen (RCC) by histomorphomertry, and level of malon-dialdehyde (MDA) and superoxide dismutase (SOD) in liver tissue by thiobarbituric acid reaction and pyrogallic acid self-oxidation, respectively. Serum albumin (ALB), alanine transaminase (ALT) and portal vein pressure (PVP) were also examined. RESULTS: rhGH up-regulated both the GH-binding capacity (R(T)) and the expression of GHR mRNA in vivo. R(T) in group A (72+/-12 fmol/mg protein) was significantly higher than that in group C (31+/-4 fmol/mg protein) (P<0.05). R(T) in group B (80+/-9 fmol/mg protein) increased markedly compared to group A (P<0.05). R(T) in group D (40+/-7 fmol/mg protein) raised remarkably compared with group C (P<0.05), but less than that in group A, and there was no significant GH binding affinity contrast (Kd) change. The GHR mRNA level (iOD, pixel) in group A (29+/-3) was significantly higher than that in group C (23+/-3) (P<0.05). GHR mRNA levels were significantly raised in group B (56+/-4) and group D (42+/-8) compared with groups A and C (29+/-3 and 23+/-3, respectively) (P<0.05). Compared with the normal liver, MDA level was higher and SOD level was lower in cirrhotic livers. After rhGH treatment, MDA level was significantly declined to 12.0+/-2.2 nmol/mg protein and SOD was raised to 1 029+/-76 U/mg protein in group D (P<0.05). ALB levels in groups B and D (42+/-7 g/L and 37+/-7 g/L, respectively) were significantly raised compared with those in groups A and C (35+/-5 g/L and 29+/-4 g/L, respectively) (P<0.05). ALT level was markedly lower in group D (69+/-7 U/L) compared to group C (89+/-15 U/L) (P<0.05), and close to group A (61+/-10 U/L). RCC in group C (22.30+/-3.86%) was significantly higher than that in group A (1.14+/-0.21%) and group D (14.70+/-2.07%) (P<0.05). In addition, rhGH markedly alleviated portal hypertension in liver cirrhotic rats (group D vs C, 9.3+/-1.5 cmH(2)O vs 14.4+/-2.0 cmH(2)O) (P<0.05). CONCLUSION: Pharmacological doses of rhGH can increase R(T) and GHR mRNA expression, ameliorate liver functions, repress fibrosis and decline portal hypertension, suggesting it has potentially clinical usage as a hepatotropic factor.

Expression of growth hormone receptor and its mRNA in hepatic cirrhosis.

AIM: To investigate the expression of growth hormone receptor (GHR) and mRNA of GHR in cirrhotic livers of rats with the intension to find the basis for application of recombinant human growth hormone (rhGH) to patients with liver cirrhosis. METHODS: Hepatic cirrhosis was induced in Sprague-Dawley rats by administration of thioacetamide intraperitoneally for 9-12 weeks. Collagenase IV was perfused in situ for isolation of hepatocytes. The expression of GHR and its mRNA in cirrhotic livers was studied with radio-ligand binding assay, RT-PCR and digital image analysis. RESULTS: One class of specific growth hormone-binding site, GHR, was detected in hepatocytes and hepatic tissue of cirrhotic livers. The binding capacity of GHR (R(T), fmol/mg protein) in rat cirrhotic liver tissue (30.8+/-1.9) was significantly lower than that in normal control (74.9+/-3.9) at the time point of the ninth week after initiation of induction of cirrhosis (n=10, P<0.05), and it decreased gradually along with the accumulation of collagen in the process of formation and development of liver cirrhosis (P<0.05). The number of binding sites (X10(4)/cell) of GHR on rat cirrhotic hepatocytes (0.86+/-0.16) was significantly lower than that (1.28+/-0.24) in control (n=10, P<0.05). The binding affinity of GHR among liver tissue, hepatocytes of various groups had no significant difference (P>0.05). The expression of GHR mRNA (riOD, pixel) in rat cirrhotic hepatic tissues (23.3+/-3.1) was also significantly lower than that (29.3+/-3.4) in normal control (n=10, P<0.05). CONCLUSION: The growth hormone receptor was expressed in a reduced level in liver tissue of cirrhotic rats, and lesser expression of growth hormone receptors was found in a later stage of cirrhosis. The reduced expression of growth hormone receptor was partly due to its decreased expression on cirrhotic hepatocytes and the reduced expression of its mRNA in cirrhotic liver tissue.

[Expression of growth hormone receptor in hepatocellular carcinoma and its significance].

BACKGROUND & OBJECTIVE: Recombinant human growth hormone (rhGH) has been proved effective in clinic, such as promoting protein synthesis and decreasing the mortality. But there are still many arguments on whether it can be used in hepatocellular carcinoma (HCC) patients. rhGH cannot work except that it combines to its own receptor growth hormone receptor (GHR). The current study was designed to explore the expression of GHR in HCC tissues and to investigate the viability of rhGH for HCC treatment. METHODS: Radioreceptor assays were used to determine growth hormone receptor (GHR) in 40 HCC tissues; 6 normal liver tissues were used as control. Receptor binding capacity(RT) and affinity constant(Kd) of GHR were calculated by Scatchard's method; the relationship between RT of GHR in cancer and clinicopathologic factors were also analyzed. RESULTS: The growth hormone- specific singular binding site, namely GHR, was detected respectively in 35 HCC cases and control tissues. The RT and Kd of GHR were 18.5416+/-4.1686 fmol/mg protein and 0.6319+/-0.1978 nmol/L in tumor tissues, 39.5467+/-3.4770 fmol/mg protein and 0.6167+/-0.1007 nmol/L in control tissues, respectively. Compared with the normal liver tissue, RT of GHR in tumor was lower (P< 0.05) but Kd did not show any difference(P >0.05). The RT of the GHR was negatively relevant to the tumor size and disease stage, but was not associated with differentiation of tumor, ages of the patients or whether the patient suffered from cirrhosis at the same time. GHR was undetectable in 5 cases. CONCLUSION: This study showed that most of the HCC tissues express low levels of GHR. Before their functions are well understood, rhGH should be very carefully used in HCC patients.

[Expression of growth hormone receptor mRNA in hepatocellular carcinoma and matched para-cancer cirrhotic liver tissue].

BACKGROUND & OBJECTIVE: The growth hormone receptor mRNA is abundant in normal liver tissue, but its expression was not studied in detail in the tissue of liver cancer or para-cancer cirrhotic liver. This study was designed to investigate the expression of growth hormone receptor mRNA in hepatocellular carcinoma and para-cancer cirrhotic liver tissue. METHODS: The expression of growth hormone receptor mRNA was detected in 37 specimens of hepatocellular carcinoma and their para-cancer liver tissue by RT-PCR. RESULTS: The expression rate of growth hormone receptor mRNA in hepatocellular cancerous tissue (30/37, 81%) was significantly lower than that in cirrhotic liver tissue (32/32, 100%) (P < 0.05). The expression rate in undifferentiated hepatocellular carcinoma (1/4, 20%) decreased further (P < 0.05). The expression of growth hormone receptor mRNA in cirrhotic liver tissue [ratio of integrated Optical Density (riOD) 30.77% +/- 8.24%, n = 32] was significantly lower than that in normal control liver tissue (riOD 44.93% +/- 6.25%, n = 5) (P < 0.05). The expression in severe cirrhotic liver tissue (riOD 21.90% +/- 4.72%, n = 8) decreased further (P < 0.05). CONCLUSIONS: The down-regulated expression of growth hormone receptor mRNA was related to the pathological stage of hepatocellular cancer and the stage of liver cirrhosis.