To observe changes of sensitivity to 5'-deoxy-5-fluorouridine (5'-dFUrd), and 5-fluorouracil (5-FU) in SMMC-7721 hepatocellular carcinoma cells by interferon alpha (IFN-alpha), and its relationship ...with the expression of thymidine phosphorylase (TP).
TP mRNA expression was determined by RT-PCR. Cytotoxicity of 5'-dFUrd, and 5-FU against SMMC-7721 cells was evaluated by MTT assay.
Expression levels of TP mRNA was elevated in SMMC-7721 cells after cultured with IFN-alpha, which was a concentration- and time-dependent increase. After SMMC-7721 cells was treated in the concentration of 5000 U/ml, and 10 000 U/ml IFN-alpha, the level of TP mRNA was significantly higher than that in untreated control (P < 0.05). When SMMC-7721 cells was cultured with the concentration 10 000 U/ml IFN-alpha, the level of TP mRNA rose at 8hr, reached the peak value at 12hr, and remained high level up to 72hr. IFN-alpha enhanced the sensitivity of SMMC-7721 cells to 5'-dFUrd with dose-dependent increase. IFN-alpha (10 000 U/ml) reduce
Purpose: This study was to investigate the effect of capecitabine on recurrent tumor and metastasis after curative resection of liver
cancer, xenograft of a highly metastatic human hepatocellular ...carcinoma (HCC) tumor (LCI-D20), with special reference to the
expression of platelet-derived endothelial cell growth factor (PD-ECGF).
Experimental Design: LCI-D20 and LCI-D35 (a low metastatic human HCC model) liver tumors were orthotopically implanted in 96 nude mice and divided
into a treatment group (24 LCI-D20 mice and 24 LCI-D35 mice) and a prevention group (48 LCI-D20 mice). In the prevention group,
curative resection of liver tumors was done 10 days after the orthotopic implantation of LCI-D20 tumor. Arabic gum (control),
5-fluorouracil (5-FU), and capecitabine were administrated respectively to all of the 96 mice.
Results: In the treatment group, tumor volume was 468 ± 138, 442 ± 81, and 240 ± 119 mm 3 ( P < 0.01) in the control, 5-FU, and capecitabine subgroups, respectively, in LCI-D20 mice, whereas it was 168 ± 35, 164 ± 23,
and 144 ± 21 mm 3 ( P > 0.05), respectively, in LCI-D35 mice. In the prevention group, incidence of liver recurrence in the control, 5-FU, and
capecitabine subgroups was 100, 100, and 50%; lung metastasis being 100, 100, and 17%; and life span being 31 ± 5, 37 ± 5,
and 77 ± 19 days, respectively. PD-ECGF was highly expressed in HCC and its metastatic tissues in LCI-D20 mice and hardly
expressed in HCC tissues in LCI-D35 mice.
Conclusions: Capecitabine inhibits tumor growth and metastatic recurrence after resection of HCC in highly metastatic nude mice model.
The effect of capecitabine may be attributed to the high expression of PD-ECGF in tumors.
Aims
To investigate the most suitable housekeeping genes for quantifying a change in mRNA expression levels due to hepatitis virus B related hepatocellular carcinoma (HCC).
Methods
Expression of mRNA ...encoding ACTB, GAPDH, B2M, HPRT and TBP was measured using quantitative reverse transcription polymerase chain reaction (qRT-PCR) in matched malignant and non-malignant tissues obtained from 65 non-treated HCCs. The software programs geNorm and NormFinder were used to ascertain the most suitable reference gene combination.
Results
All candidate genes showed significantly different expression between malignant and non-malignant samples. GAPDH and ACTB, genes most frequently used for normalization, were heavily regulated during HCC carcinogenesis and progression. B2M expression levels varied with hepatitis infection status. The combination of HPRT and TBP expression levels were the most stable, regardless of differences in tumor stage and cirrhotic and malignancy status.
Conclusions
It is necessary to select reference genes based on tissue and disease specific expression profile and to further identify novel reference genes with greater expression stability for use in HBV related HCC gene expression studies.
To analyze the risk factors influencing the prognosis of orthotopic liver transplantation for hepatocellular carcinoma (HCC) and sum up the relevant clinical experience in diagnosis and treatment of ...HCC.
The clinical data of 198 HCC patients, 177 males and 21 females, aged 49 (24-83), were analyzed.
The 0.5-, 1-, and 2-year survival rates were 89%, 78%, and 65 respectively. The rates of disease-free survival (DFS) were 85%, 73, and 67% respectively. Univariate analysis revealed that tumor size, presence of vascular invasion, Edmondson grade, TNM classification, and preoperative alpha-fetoprotein (AFP) were significantly related to DFS, and the 4 foregoing factors were also related to the survival rate. Cox regression analysis suggested that presence of vascular invasion was an independent prognostic factor of survival rate and DFS.
Vascular invasion plays a leading role in evaluating the prognosis of orthotopic liver transplantation for HCC. It is important to discover the micro-metastasis and explore more ef
To explore the diagnosis and management of early hepatic artery thrombosis (HAT) after liver transplantation.
Routine examination of Color Doppler Imagine (CDI) was used to detect hepatic artery flow ...after liver transplantation in 220 cases from April 2001 to November 2004. Suspected patients were further confirmed by immediate hepatic artery angiography, and continuous infusion of urokinase through hepatic artery with catheter was performed to the patients with HAT.
HAT was identified in 6 patients (2.7%), occurring 5.5 days (2 - 19 days) after liver transplantation. Hepatic artery recanalization was obtained in 6 cases. One patient died from lung infection 2 months after liver transplantation. One patient underwent the second liver transplantation because of the recurrence of HAT 6 months after the first transplantation, but died from multiple system organ failure 2 months after the operation. The other 4 cases have been surviving well disease-freely.
CDI is useful for the diagnosis of early HAT after liver
To investigate the antitumor effect of interferon-alpha 2a (IFN-alpha2a) combined with capecitabine on hepatocellular carcinoma (HCC) in nude mice in relation to thymidine phosphorylase (TP) ...expression.
Thirty nude mice bearing orthotopic xenografts of a human HCC tumor (LCI-D20) were divided into control, capecitabine, IFN-alpha2a, and combination (capecitabine plus IFN-alpha2a) groups. Tumor growth was determined by measuring the tumor volume. An enzyme-linked immunosorbent assay (ELISA) was used to study the TP expression in the cancer tissues of the liver.
IFN-alpha2a enhanced the sensitivity of the LCI-D20 tumor response to capecitabine treatment. The tumor volume was significantly reduced in the capecitabine (455+/-236 mm(3)), IFN-alpha2a (248 +/- 114 mm(3)) or combination (46 +/- 29 mm(3)) treatment groups as compared to the control (1,033 +/- 146 mm(3)) (P < 0.01). A significant difference was also found between the single treatment (capecitabine or interferon) and combination treatment group (P < 0.01 and P < 0.05, respectively). IFN-alpha2a up-regulated TP expression in LCI-D20 tumor. An approximate 1.5-fold increase in TP expression was observed in the mice which received IFN-alpha2a treatment compared to the control mice.
IFN-alpha2a enhanced the antitumor effect of capecitabine on HCC in nude mice, which might be ascribed to the up-regulation of TP expression in liver cancer tissues by IFN-alpha2a.
To further study the impact of interferon-alpha (IFN-alpha) on thymidine phosphorylase (TP) expression and angiogenesis.
Human hepatocellular carcinoma cells of the line SMMC-7721 were cultured and ...added with IFN-alpha of different doses: 0 (as control group), 1000, 5000 and 10,000 U/ml. Twenty-four hours later RT-PCR was used to detect the TP mRNA expression. Boyden chamber method was used to examine the endothelial cells migration. Suspension of SMMC-7721 cells was inoculated subcutaneously into 30 male BALB/c-nu/nu mice, the mice were randomly divided into 5 equal groups to be subcutaneously injected with IFN-alpha of different doses: 0 (as control group), 1.0 x 10(6), 3.0 x 10(6), 9.0 x 10(6), and 1.5 x 10(7) U.kg(-1).d(-1) for 3 weeks. The eating behavior, activity, body weight, and tumor size were observed. The rats were killed 2 days after the drug injection was stopped. The subcutaneous tumors were taken out to undergo histological examination and TP protein expression by ELISA. The microvessel densit
Background Selection of patients with hepatocellular carcinoma (HCC) for orthotopic liver transplantation (OLT) remains controversial. Since there is a trend to expand the transplant criteria for HCC ...patients, we reviewed the data of patients with HCC who had received OLT at our institute to determine their survival and prognostic factors. Methods A total of 67 patients with HCC who had undergone OLT from April 2001 through December 2003 were reviewed retrospectively. Selection OLT candidates with HCC was dependent on the anatomical characteristics and/or the severity of underlying liver cirrhosis. The 67 patients were followed up for more than 6 months after transplantation. Their survival rate was calculated by the Kaplan-Meier method. Univariate and multivariate analyses using the Cox proportional hazards regression model were performed to reveal the factors affecting the survival rate. Results No perioperative death occurred in this series. The 1- and 2-year cumulative survival rates were 90.0% and 65.6%, and the disease-free survival (DFS) rates were 77.5% and 62.5% respectively. Univariate analysis revealed the tumor size, portal vein tumor thrombus (PVTI'), serum alpha-fetoprotein level, bilobular distribution of tumors, pTNM stage and histological differentiation were statistically significant factor saffecting the DFS (P < 0.05 ). Multivariate analysis showed tumor size and PVTT were independent and statistically significant factors affecting the DFS (P =0. 005 and 0. 010, respectively). In this series, all but 2 received systemic chemotherapy, among them 13 had tumor recurrence within 8 months after OLT. Conclusions OLT is indicated for patients with HCC, even for some patients with end-stage liver disease who may survive longer without tumor recurrence. Adjuvant chemotherapy may decrease the recurrence of HCC after OLT.
To investigate the effects on sensitivity to fluoropyrimidine and endothelial cell (EC) migration by transfection with thymidine phosphorylase (TP) cDNA to a hepatocellular carcinoma (HCC) cell line ...SMMC-7721.
SMMC-7721 was transfected with pcDNA3.1/zeo (+) with human TP cDNA. TP mRNA expression was determined by RT-PCR. Sensitivity to fluoropyrimidine was determined by 3-4,5-dimethylthiazol-2-yl-2, 5-diphenyl-tetrazolium bromide (MTT) assay. Induction of EC migration was detected by Boyden chamber assay.
The construction of pcDNA3.1/zeo(+)-TP was verified by digestion with restriction endonuclease Apa1. When comparison was made between SMMC-7721 cell clone transfected with pcDNA3.1/zeo(+)-TP (S-TP) and control clone transfected with pcDNA3.1/zeo(+) (S-vector), we found that TP mRNA expression level was much higher in S-TP, being 2.09+/-0.16 vs 0.48+/-0.06 in S-vector (P < 0.01), sensitivity to 5'-deoxy-5-fluorouridine (5'-dFUrd, a prodrug of 5-fluorouracil) in S-TP was significantly enhanced compared with that in S-vector (IC(50); 56.81+/-9.85 micromol/l vs 162.25+/-11.03 micromol/l, P < 0.01), and the culture medium of S-TP possessed more potential to induce EC migration than that of S-vector (the number of ECs appearing on the outer surfaces of the membrane was 275+/-29 vs 122+/-35 per field, P < 0.01).
Sensitivity to 5'-dFUrd could be enhanced by transfection with TP cDNA for SMMC-7721 cells. However, EC migration was also promoted at the same time. Therefore, transfection with TP alone might have no potential to enhance anti-tumoral effects of fluoropyrimidine in HCC.
