Decreased expression of ApoF associates with poor prognosis in human hepatocellular carcinoma.

BACKGROUND: Hepatocellular carcinoma (HCC) is frequently associated with metabolism dysfunction. Increasing evidence has demonstrated the crucial role of lipid metabolism in HCC progression. The function of apolipoprotein F (ApoF), a lipid transfer inhibitor protein, in HCC is incompletely understood. We aimed to evaluate the functional role of ApoF in HCC in this study. METHODS: We used quantitative reverse-transcription polymerase chain reaction (qRT-PCR) to detect ApoF mRNA expression in HCC tissues and hepatoma cell lines (SMMC-7721, HepG2, and Huh7). Immunohistochemistry was performed to detect the expression of ApoF in HCC tissues. The associations between ApoF expression and clinicopathological features as well as HCC prognosis were analyzed. The effect of ApoF on cellular proliferation and growth of SMMC-7721 and Huh7 cells was examined in vitro and in vivo. RESULTS: ApoF expression was significantly down-regulated at both mRNA and protein levels in HCC tissues as compared with adjacent tissues. In SMMC-7721 and Huh7 HCC cells, ApoF overexpression inhibited cell proliferation and migration. In a xenograft nude mouse model, ApoF overexpression effectively controlled HCC growth. Kaplan-Meier analysis results showed that the recurrence-free survival rate of HCC patients with low ApoF expression was significantly lower than that of other HCC patients. Low ApoF expression was associated with several clinicopathological features such as liver cirrhosis, Barcelona Clinic Liver Cancer stage and tumor-node-metastasis stage. CONCLUSIONS: ApoF expression was down-regulated in HCC, which was associated with low recurrence-free survival rate. ApoF may serve as a tumor suppressor in HCC and be a potential application for the treatment of this disease.

Circular RNA circCDK13 suppresses cell proliferation, migration and invasion by modulating the JAK/STAT and PI3K/AKT pathways in liver cancer.

Circular RNAs have recently been disclosed as potential biomarkers for human cancers. This study aimed to characterize the expression and function of a novel circular RNA, circCDK13, in liver cancer progression, as well as to elucide the underlying mechanisms. For this purpose, circCDK13 expression was quantitatively analyzed by RT-PCR in various liver cancer cell lines and human cancerous tissues. The migration, cell cycle progression, proliferation and invasion of liver cancer cells with an enhanced circCDK13 expression were evaluated by 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium, inner salt (MTS) assay, flow cytometry and the Transwell culture system. Microarray and western blot analyses were performed to explore the underlying signaling mechanisms. The role of circCDK13 in liver cancer was finally examined by tumorigenicity assay using nude mice. The results revealed that circCDK13 expression was suppressed in various liver cancer lines and tissue samples from patients with liver cancer (hepatocellular carcinoma). The induced overexpression of circCDK13 in the liver cancer cells markedly inhibited their migration rates, altered cell cycle progression, and suppressed the cell migratory and invasive capacities. Microarray analysis also identified numerous downstream genes regulated by circCDK13, particularly those in the Janus tyrosine kinase (JAK)/signal transducer and activator of transcription (STAT) and phosphoinositide 3-kinase (PI3K)/AKT signaling pathways. The results of the tumorigenicity assay revealed that circCDK13 overexpression significantly inhibited liver cancer progression in nude mice. On the whole, the findings of this study indicate that circCDK13 is a novel circular RNA that suppresses liver cancer progression, and that these suppressive effects are possibly mediated via the JAK/STAT and PI3K/AKT signaling pathways.

Evaluation of hepatic perfusion and function with modified hepatobiliary scintigraphy.

The quantitative relationship between changes in portal vein pressure and the perfusion index was studied, and hepatocellular function was evaluated. A modified protocol of hepatobiliary dynamic scintigraphy with Tc-99m-labeled ethylene hepatobiliary iminodiacetic acid (EHIDA) was performed in 37 patients with hepatic cirrhosis and 12 healthy controls. In 18 patients, the portal vein pressure (PVP) was measured intra-operatively during the portal vein bypass procedure. The portal vein perfusion index (PVI) was obtained by a two-compartment model of hepatic perfusion. A three-compartment model was applied in the hepatocellular extracted and excreted time-radioactivity curves, and the hepatic functional index was calculated including the hepatic uptake index (UI), the mean residual index (MRI), the uptake speed index (UsI), the descent speed index (DsI), the peak uptake (PU), and the peak time (PT) of hepatic uptake. These function indices in the cirrhotic patients were compared to the indices in the healthy controls with a two-sided t-test; specifically, the PU (18.94+/-6.80 vs. 29.67+/-18.98, P<0.01), UI (0.84+/-5.99 vs. 18.41+/-13.87, P<0.05), and UsI (0.58+/-0.39 vs. 1.23+/-0.81, P<0.01) were slower in patients with hepatic cirrhosis compared with the healthy controls. The PT (16.83+/-4.89 vs. 10.95+/-2.79, P<0.001) and MRI (5.93+/-2.96 vs. 2.74+/-0.97, P<0.001) were more prolonged and the DsI (0.07+/-0.09 vs. 0.18+/-0.14, P<0.01) was less compared to controls. The PVI were significantly higher in cirrhotic patients than in controls (46.17+/-7.83 vs. 29.07+/-5.71, P<0.001), and the PVI correlated to the PVP (r=0.79, P<0.01). In conclusion, modified quantitative hepatobiliary dynamic scintigraphy can evaluate the changes of portal vein blood flow and monitor hepatocellular function, in which the portal vein blood flow can estimate portal vein pressure.

Glutamine and recombinant human growth hormone protect intestinal barrier function following portal hypertension surgery.

AIM: To evaluate the effects of combined treatment of glutamine (Gln) and recombinant human growth hormone(rhGH) on intestinal barrier function following portal hypertension surgery. METHODS: This study was designed as a prospective, randomized and controlled clinical trial. Forty two patients after portal hypertension surgery were randomly assigned into 2 groups: control group (n = 20) and supplemental group (adding Gln and rhGH, n = 22). Every patient received isocaloric and isonitrogenous standard total parenteral nutrition (TPN) starting 3 d after surgery for 7 d. Blood samples were obtained before surgery and at the 3rd and 10th day postoperatively. Host immunity was evaluated by measuring levels of CD4, CD8, CD4/CD8, IgG, IgM and IgA, and the inflammatory responses were determined by assessing IL-2, TNF-alpha and C-reactive protein (CRP) levels. Intestinal permeability and integrity was evaluated by L/M test and histological examination, respectively. RESULTS: On postoperative d 10, CD4, CD4/CD8, IgG and IL-2 levels in supplemental group were significantly higher than those in control group (33.7 +/- 5.5 vs 31.0 +/- 5.4, P < 0.05, (1.17 +/- 0.32 vs 1.05 +/- 0.15, P < 0.05, 13.94 +/- 1.09 vs 12.33 +/- 1.33, P < 0.05, and 368.12 +/- 59.25 vs 318.12 +/- 45.65, P < 0.05, respectively), whereas the increase in serum TNF-alpha concentration was significantly reduced (41.02 +/- 27.56 vs 160.09 +/- 35.17, P < 0.05). The increase in L/M ratio was significantly lower in the supplemental group than in the control group (0.0166 +/- 0.0017 vs 0.0339 +/- 0.0028, P < 0.05). Moreover, mucosal integrity in the supplemental group was better than in the control group. CONCLUSION: Postoperative administration of TPN supplemented with Gln and rhGH in patients after portal hypertension surgery improves immune function, modulates inflammatory response, prevents the intestinal mucous membrane from atrophy and preserves intestinal integrity.

Evaluation of the effects of combined endoscopic variceal ligation and splenectomy with pericardial devascularization on esophageal varices.

AIM: To detect the hemodynamic alterations in collateral circulation before and after combined endoscopic variceal ligation (EVL) and splenectomy with pericardial devascularization by ultrasonography, and to evaluate their effect using hemodynamic parameters. METHODS: Forty-three patients with esophageal varices received combined EVL and splenectomy with pericardial devascularization for variceal eradication. The esophageal vein structures and azygos blood flow (AZBF) were detected by endoscopic ultrasonography and color Doppler ultrasound. The recurrence and rebleeding of esophageal varices were followed up. RESULTS: Patients with moderate or severe varices in the esophageal wall and those with severe peri-esophageal collateral vein varices had improvements after treatment, while the percentage of patients with severe para-esophageal collateral vein varices decreased from 54.49% to 2.33%, and the percentage of patients with detectable perforating veins decreased from 79.07% to 4.65% (P < 0.01). Color Doppler flowmetry showed a significant decrease both in AZBF (43.00%, P < 0.05) and in diameter of the azygos vein (28.85%, P < 0.05), while the blood flow rate was unchanged. The recurrence rate of esophageal varices was 2.5% (1/40, mild), while no re-bleeding cases were recorded. CONCLUSION: EVL in combination with splenectomy with pericardial devascularization can block the collateral veins both inside and outside of the esophageal wall, and is more advantagious over splenectomy in combination with pericardial devascularization or EVL in preventing recurrence and re-bleeding of varices.

[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.