ABSTRACT
ObjectiveTo investigate the potential mechanism of serum N-glycan alterations in patients with hepatitis B virus (HBV)-related hepatocellular carcinoma (HCC) by measuring serum N-glycan profile and comparing glycosyltransferase gene expression between HCC tissue and adjacent tissue. MethodsThe samples of HCC tissue, adjacent tissue, and normal liver tissue were collected from 34 patients with HBV-related HCC who were admitted to Chinese PLA General Hospital, and serum samples were also collected. Among these 34 patients, 8 were randomly selected and their serum samples were established as HCC experimental group, and the serum samples of 20 healthy adults were established as control group. DNA sequencer-aided fluorophore-assisted carbohydrate electrophoresis was used to analyze serum N-glycan profile in the HCC experimental group and the control group. Quantitative real-time PCR was used to measure the mRNA expression of 8 glycosyltransferase genes (FUT3, FUT4, FUT6, FUT7, FUT8, Gn-TIII, Gn-TIVa, and Gn-TV) in the HCC tissue and adjacent tissue of 34 patients with HBV-related HCC, and Western blot was used to measure the expression of corresponding proteins. The independent samples t-test was used for comparison of continuous data between two groups. ResultsCompared with the control group, the HCC experimental group had a significant increase in the abundance of N-glycan peak9 (NA3Fb) in serum(t=-2.514,P＜0.05). There were significant differences in the mRNA expression of FUT8, Gn-TIVa, and Gn-TV between HCC tissue and adjacent tissue, and the mRNA and protein expression levels of FUT8 and Gn-TV in HCC tissue were significantly higher than those in adjacent tissue (FUT8 mRNA: 1.50±0.34 vs 0.65±0.11, t=-2.354，P=0.022; Gn-TV mRNA: 3.57±0.64 vs 1.33±016, t=-3.384,P=0001; FUT8 protein: 0.70±0.11 vs 0.083±0.017, t=9.555,P=0.001; Gn-TV protein: 1.33±0.19 vs 0.60±0.15, t=5.097,P=0.007). The mRNA expression level of Gn-TIVa in HCC tissue was significantly higher than that in adjacent tissue (2.90±0.47 vs 1.68±0.19, t=-2.403,P=0.019), but there was no significant difference in the protein expression level of Gn-TIVa between HCC tissue and adjacent tissue (052±0.24 vs 0.24±0.11,t=1.833, P=0.141). The changes of glycosyltransferase gene expression in HCC tissue were consistent with the alteration of serum N-glycan profile. ConclusionSerum N-glycan alterations in patients with HBV-related HCC may be closely associated with the upregulated expression of the glycosyltransferase genes FUT8, Gn-TIVa, and Gn-TV in HCC tissue.
ABSTRACT
ObjectiveTo investigate the effect of hypoxia-inducible factor-1α (HIF-1α) on the stemness and epirubicin sensitivity of hepatoma cells. MethodsHepatoma cells were selected for experiment. HepG2 hepatoma cells transfected with HIF-1α overexpression plasmid were selected as experimental group, and those transfected with pcDNA3.1 empty plasmid were selected as control group; HepG2 cells alone were selected as HepG2 group. Quantitative real-time PCR was used to measure the mRNA expression of HIF-1α; Western blot was used to measure the protein expression of HIF-1α; flow cytometry was used to measure the expression of CD133 on the surface of hepatoma cells. The three groups of cells were treated with epirubicin at different concentrations (0, 6.25, 12.5, 25, and 50 μmol/L) for 24 hours; MTT assay was used to measure cell viability, and flow cytometry was used to measure apoptosis after treatment with epirubicin (50 μmol/L). A one-way analysis of variance was used for comparison of continuous data between multiple groups, and the t-test was used for further comparison between two groups. ResultsCompared with the HepG2 group and the control group, the experimental group had a significant increase in the mRNA expression of HIF-1α (both P＜0.001), and Western blot showed high expression of HIF-1α in the experimental group. The percentage of CD133 cells was 0.040%±0.003% in the HepG2 group, 0.030%±0.010% in the control group, and 20.110%±0.600% in the experimental group, and the experimental group had a significantly higher positive rate of CD133+ than the HepG2 group and the control group (both P＜0.001). At an epirubicin concentration of 25 and 50 μmol/L, the HepG2 group and the control group had significantly inhibited cell viability and a significantly lower cell viability than the experimental group (both P＜005). After the treatment with 50 μmol/L epirubicin for 48 hours, the experimental group had a significantly lower cell apoptosis rate than the HepG2 group (67.9%±2.5% vs 93.6%±1.5%, P＜0.001) and the control group (67.9%±2.5% vs 93.0%±1.2%, P＜0001). ConclusionHepG2 cells are successfully transfected with HIF-1α overexpression plasmid, and HIF-1α can increase the percentage of liver cancer stem cells and improve their resistance to epirubicin.
ABSTRACT
Objective To investigate the effect of atractylone on the viability and apoptosis of hepatoma HepG2 cells and its mechanism of action. Methods Hepatoma HepG2 cells were selected and divided into low-, middle-, and high-dose atractylone groups (5, 10, and 20 μmol/L), and the cells in the control group were added with an equal volume of DMSO. MTT colorimetry was used to measure the viability of HepG2 cells after treatment with different concentrations of atractylone; flow cytometry was used to measure the apoptosis rate and mitochondrial membrane potential of HepG2 cells; the DCFH-DA fluorescent probe labeling method was used to measure the level of reactive oxygen species (ROS) in HepG2 cells; Transwell assay was used to evaluate the effect of atractylone on the migration ability of HepG2 cells; Western blot was used to measure the protein expression levels of Bcl-2, Bax, and cleaved caspase-3. A one-way analysis of variance was used for comparison of continuous data between multiple groups, and the least significant difference t -test was used for comparison between two groups. Results After 24 and 48 hours of treatment with atractylone, compared with the control group, the low-, middle-, and high-dose atractylone groups had a tendency of reduction in cell viability (all P < 0.05), with a half inhibitory concentration of 26.19 μmol/L in atractylone treatment of HepG2 cells for 72 hours. The low-, middle-, and high-dose atractylone groups had a significantly higher apoptosis rate than the control group (14.34%/29.32%/50.12% vs 0.32%, all P < 0.05). Compared with the control group, the low-, middle-, and high-dose atractylone groups had a significant increase in the fluorescence intensity of ROS in HepG2 cells (all P < 0.05). After 48 hours of treatment with atractylone, compared with the control group, the low-, middle-, and high-dose atractylone groups had a significant reduction in the number of migrated cells (132.67±18.36/57.00±9.26/31.00±2.45 vs 258.11±38.54, P < 0.05). Compared with the control group, the low-, middle-, and high-dose atractylone groups had a significant reduction in the expression of the anti-apoptotic factor Bcl-2 and significant increases in the expression of the apoptotic factors Bax and cleaved caspase-3 (all P < 0.05). Conclusion Atractylone can induce the apoptosis and inhibit the migration of HepG2 cells, which provides an experimental basis for further development and utilization of atractylone.
ABSTRACT
Objective: To explore the effect of transcatheter embolization combined with radiofrequency ablation (RFA) in treatment of VX2 liver tumors of rabbits. Methods: Sixty rabbit VX2 liver tumor models were randomly divided into 4 groups (each n=15). Fifteen minutes after TACE or transcatheter arterial embolization (TAE), RFA was performed in both TACE+RFA group and TAE+RFA group, respectively.And RFA group was only treated with RFA, TACE group underwent only TACE. Serum aspartate aminotransferase (AST) and alanine aminotransferase (ALT) were measured 1 day before and 3 and 7 days after operation. Tumor growth rate, tumor necrosis rate and Suzuki score were measured 7 days after operation. The expression of heat shock protein 70 (HSP70) in liver lesions surrounding ablation zone or embolized zone was detected with immunohistochemistry 1, 3 and 7 days after operation, and hepatocyte apoptosis index and proliferation index were calculated. Results: The levels of ALT and AST in TACE+RFA group were higher than those in other groups 3 and 7 days after operation (all P<0.05). Seven days after operation, the Suzuki score in TACE+RFA group was higher than that in other groups, and tumor growth rates in TACE+RFA and TAE+RFA groups significantly decreased, while the necrosis rates in TACE+RFA and TAE+RFA groups increased compared with RFA and TACE groups (all P<0.05). The expression of HSP70 in hepatic lesions surrounding the ablation zone or embolized zone increased gradually 1, 3 and 7 days after operation, and in TACE+RFA group were higher than in other groups, in TAE+RFA group were higher than in TACE group and RFA group 3 days after operation (all P<0.05). The hepatocyte apoptosis index in hepatic lesions surrounding the ablation zone or embolized zone decreased gradually in all 4 groups 1, 3 and 7 days after operation, while in TACE+RFA group were higher than the other 3 groups, in TACE group 1 and 3 days after operation were higher than in TAE+RFA group and RFA group (all P<0.05). The hepatocyte proliferation index in 4 groups were higher 3 days after operation than that 1 and 7 days after operation, and compared with other groups, in TAE+RFA group were higher 1, 3 and 7 days after operation, in RFA group was higher 1 and 3 days after operation than in TACE+RFA group and TACE group (all P<0.05). Conclusion: The effects of TACE+RFA and TAE+RFA on inhibiting tumor growth are better than TACE and RFA alone for treatment of VX2 liver tumors of rabbits. Compared with TACE+RFA, TAE+RFA can promote hepatocyte proliferation and inhibit hepatocyte apoptosis more effectively with less liver damage.
ABSTRACT
Objective To study pharmacokinetics and tissue distribution of CalliSpheres Beads (CB) loaded Arsenic trioxide (ATO) on rabbit VX2 liver tumor by transcatheter arterial chemoembolization (TACE). Method Sixty four rabbits with VX2 liver tumors were randomly divided into 4 groups: control group, CB group, CBATO group and cTACE group. Blood samples were taken at specific time points after TACE.The blood concentration of ATO,liver and kidney functions were examined respectively. In each group, every 4 rabbits were sacrificed on 1 days,3 days,7 days and 14 days after operation. The tumor,liver,kidney, lung,heart and muscle were taken to detect the drug concentration. Bilateral t?test was used to compare the drug concentration in blood and tissue between CBATO group and cTACE group. Results Statistically,The levels of ALT and AST in group CBATO and cTACE on 1st,3rd and 7th days after TACE were significantly higher than those in CB group(ALT: F=25.872, 17.69, 7.016, AST: F=46.365, 32.385, 12.548, P<0.05) respectively. The ALT and AST levels in CBATO group were statistically lower than those in cTACE group (ALT: t=0.369, 0.432, 0.169, 0.353, AST: t=0.488, 0.593, P>0.05). There were no statistically significant differences in the levels of BUN and Scr between the four experimental groups at each observation time point. Statistically, 10 minutes and 20 minutes after TACE, the blood drug concentration inCBATO was significantly lower than that in cTACE (t=7.675, 6.461, P<0.001). while 12 hours after operation,blood drug concentration in CBATO group was higher than that in cTACE group. In tumor tissue,the concentration of ATO in CBATO was higher than that in cTACE,and there was no statistical differences on the 1st day after TACE(t=2.155, P=0.068), but there was a statistical differences between 3rd, 7th and 14th days (t=11.462, 7.624, 2.649, P<0.05). Conclusion CBATO could prolong the time of drug metabolism,increase the drug concentration in tumor tissue,and didn′t aggravate the damage of liver and kidney function.
ABSTRACT
Objective To determine whether low frequency ultrasound mediated microbubbles destruction (UMMD) could inhibit VX2 orthotopic hepatic tumor growth in rabbit models.Methods Twenty-four New Zealand white rabbits were implanted with VX2 tumor in left hepatic lobe to establish a homograft rabbit model of liver neoplasms in situ,which were randomly divided into four groups(6 rabbits in each group):group A (intravenous saline only),group B (intravenous microbubbles only),group C (intravenous saline+ low frequency focused ultrasound exposure),and group D (intravenous microbubbles+low frequency focused ultrasound exposure).After 3 days consecutive treatment,tumor volume(TV),and peak intensity (PI) were monitored by conventional ultrasound and contrast enhanced ultrasound(CEUS) on 0,1,7,14 and 21 days after treatment.The rabbits were euthanized at the end of the experiment.Tumor tissues were evaluated by HE stain.Results The parameters of TV and PI of each tumor had no significant difference among four groups before treatment(all P>0.05).TV had no significant difference among four groups on 1 day after treatment(all P>0.05);PI in group C and group D were significantly lower than those in group A and group B (all P0.05).The pathological changes of necrosis tissue,hemorrhagic damage of microvessel and thrombosis were observed in the tumors of group D only,whereas these changes occurred rarely in other groups.Conclusions UMMD can inhibit the growth of VX2 hepatic tumors in rabbits,and be used as a promising novel therapeutic strategy to liver neoplasms.
ABSTRACT
The incidence rate of liver cancer tends to increase in recent years, and the molecular and pathogenic mechanisms of liver cancer should be further investigated to guarantee health. As an important model organism, zebrafish have highly conserved genes and grow fast. The early embryo of zebrafish is transparent, which helps with the real-time observation of the development process. Zebrafish are similar to humans in the composition, function, and signaling pathways of hepatocytes, as well as response to injury. In modern biological studies, zebrafish have been wildly used as the model of liver diseases. This article summarizes the research advances in the application of zebrafish as the model of liver cancer, and points out that the techniques for establishing the zebrafish model of liver cancer have become mature. With the constant development of experimental techniques, great achievements will be achieved in the field of liver cancer.
ABSTRACT
Objective To investigate the optimal ultrasound exposure parameter on H22 neoplasms of mice meditated by ultrasound exposure combined with self‐made nanobubbles ,and then observe their therapeutic effect combined with cisplatin and their possible mechanism of anti‐tumor . Methods Thirty mice engrafts models with subcutaneous H22 neoplasms were established and divided into 6 groups randomly ,which received ultrasound exposure at different intensity and exposure time . The contrast enhanced ultrasound imaging ( CEUS ) was performed in every group at the four time points of before treatment and at 0 h ,24 h ,72 h after treatment . To obtain the optimal ultrasound parameters ,the tumor inhibitory effect was assessed by enhanced intensity ( EI) and microvascular density ( MVD) . The H22 tumor were treated by ultrasound exposure nanobubbles combined with cisplatin to observe their tumor growth inhibition rate ,and the microvessels density and nuclear associated antigen Ki‐67 proliferation index were measured by immunohistochemical staining . Results There was a statistically difference in enhanced intensity (EI) between the experimental groups and control group ( P < 0 .05) . With the increasing of ultrasound intensity and exposure time ,the tumor inhibitory effect was more obvious ,with an increasing side reactions . Except the simple ultrasound group ,there was a statistical difference in tumor inhibition ,the mean MVD and the tumor cell proliferation index (KI‐67) between control group and the other ultrasound therapy groups ( P<0 .05) . The tumor inhibitory rate was the highest ( tumor inhibition rate 70 .0% ) and the mean MVD and KI‐67 expression were the lowest ( P <0 .05) in the combination group comparing with the others . Conclusions The ideal ultrasound exposure parameter of tumor inhibition showed that exposure intensity chose 1 W/cm2 and exposure time chose 1 min or 3 min intermittence . The ultrasound exposure self‐made nanobubbles combined with cisplatin could enhance the tumor inhibitory effect .Its mechanism may be related to the decrease of microvascular density ,the inbition of tumor cell proliferation and the increase of tumor cell necrosis .
ABSTRACT
Objective To investigate the effect of elemene on mRNA expressions of tumor necrosis factor (TNF) receptor associated factor 6 (TRAF6) and caspase-8 in tumor tissues of mice bearing hepatoma H22.Methods Forty BALB/c mice models bearing hepatoma H22 were established by subcutaneous inoculating tumor cells.Forty BALB/c mice were randomly divided into 4 groups:model group,low-and high-elemene dosage groups,and cisplatin group.The tumors after executing mice were weighted.The mRNA expressions of TRAF6 and caspase-8 in tumor tissues were detected by quantitative real-time reversetranscription polymerase chain reaction (RT-PCR).Results The dosage of elemene could inhibit tumor growth.The inhibition ratio of cancer in the low-and high-elemene dosage and cisplatin group was 24.2％,27.4％,and 28.2％,respectively.It reduced significantly tumor weights(P ＜0.01).Compared to the model group,the expression level of TRAF6 mRNA on tumors was decreased significantly,while the expression level of caspase-8 mRNA was increased significantly in the other groups(P ＜ 0.05).Conclusions The present results indicated that molecular mechanism of inhibition of liver cancer growth treated by elemene might be through down-regulating mRNA expressions of TRAF6 and caspase-8,promoting tumor cells apoptosis,and achieving the anti-tumor effect.
ABSTRACT
Objectives To explore the effects of the blood supply extent on the temperature of target site of tumor and volume of coagulation necrosis (V),and to provide experimental evidence for further precise control of dosage of high intensity focused ultrasound (HIFU) and improve the efficiency of HIFU in the treatment.Methods Twenty-four rabbit liver VX2 tumor models were established and were divided into four groups:10 d group,15 d group,20 d group,no blood supply group (rabbits were put to death in the 10 d after the models were established).The same irradiation parameters of HIFU were used to irradiate the hepatic tumor tissue of every group,the real-time temperature of target site of tumor were measured,the software from temperature controller was used to plot the time-temperature curve (TTC),V was measured after HIFU.Residual tumor tissue was resected for pathological observation and microvascular density (MVD) determination.Results (1) Histopathological analysis showed that the extent of a tumor's blood supply followed the order 10 d group ＞ 15 d group ＞ 20 d group (P ＜ 0.01).(2)Tmax,T1,k1:betweengroups had no statistical difference among 10 d group,15 d group,20 d group (P ＞0.05),T T1,and k1 between three groups with no blood supply group had statistical difference (P ＜ 0.05).(3) k2:10 d group ＞ 15d group ＞ 20d group ＞ no blood supply group;T2:10d group ＜ 15d group ＜ 20d group ＜ no blood supply group;(4)V:10 d group ＜ 15 d group ＜ 20 d group ＜ no blood supply group.Conclusions The extent of a tumor's blood supply had a significant effect on the temperature-decrease phase but not on the temperature-increase phase during HIFU treatment.The more abundant blood supply of the tumor was,the faster heat abstraction after HIFU was;and the easier the tissue return to normal temperature,the smaller the volume of coagulation necrosis tissue were.
ABSTRACT
Purpose To establish rabbit VX2 tumor model and to explore the relation between perfusion parameters and the expression of the VEGF in the portal vein VX2 implanting tumor emboli. Materials and Methods VX2 tumor was implanted in the portal vein of eight experimental rabbits. Multi-slice CT (MSCT) perfusion scan was performed after tumor formation to measure and compare portal vein tumor thrombus, hepatic blood lfow (HBF) near tumor foci and far away from tumor foci, hepatic blood volume (HBV), probability of surface area product (PS) and mean transit time (MTT). The VX2 tumor emboli were then resected to analyze the relationship between the liver perfusion parameters and VEGF expression using immunohistochemical method. Results MSCT liver perfusion parameters were not statistically signiifcant between foci close to or far away from the tumor (P>0.05). The HBF, HBV and PS within the tumor emboli were higher than that in hepatic parenchyma (P<0.05) and the MTT was higher (P<0.05). There was positive correlation (r=0.711, 0.646 and 0.626, P<0.05) between the HBF, HBV and PS of portal vein VX2 tumor emboli and VEGF expression, and there was negative correlation between MTT and VEGF expression (r=-0.565, P<0.05). Conclusion MSCT perfusion parameters in the portal vein VX2 implanting tumor emboli and the expression of VEGF are positively related. MSCT can evaluate the angiogenesis of portal vein VX2 implanting tumor emboli.
ABSTRACT
Objective To comparative analysis the rabbit VX2 tumors angiogenesis in fatty liver and normal liver,and investigate the correlation of contrast-enhanced ultrasound (CEUS) parameters and their angiogenesis.Methods Rabbit models of fatty liver and normal liver with implanted VX2 tumors were established.Two groups of hepatic backgrounds and VX2 tumors were analyzed by QontraXt quantitative software of CEUS.The expression level of tumor microvascular density (MVD) and vascular endothelial growth factor (VEGF) were detected with immunohistochemical techniques and real-time fluorescence quantitative PCR (FQ-PCR).Results ①No significant difference was found in MVD,VEGF protein and gene expression between VX2 tumors in normal liver and fatty liver.The expression of VEGF gene in fatty liver parenchyma were lower than normal liver parenchyma (P ＜0.05).②There was positive correlation between MVD and peak intensity (r =0.494,P ＜0.05; r =0.591,P ＜0.01) both in fatty liver and normal liver VX2 tumors.The expression of VEGF protein were not correlated with TIC parameters (all P ＞ 0.05).③The MVD had positive correlation with expression of VEGF protein in fatty liver and normal liver VX2 tumors (r =0.508,P ＜0.05; r =0.570,P ＜0.01).Conclusions The expression of MVD and VEGF had no significant difference between fatty liver and normal liver VX2 tumors.The peak intensity of VX2 tumors CEUS had positive correlation with MVD both in fatty liver and normal liver,which can indirectly reflect MVD expression level and help to evaluate tumor angiogenesis.
ABSTRACT
Objective: To investigate the effects of TFPI-2 (tissue factor pathway inhibitor 2) on the growth and angiogenesis of subcutaneous tumor xenografts in nude mice established from human liver cancer cells. Methods: The Hep3B cells stably expressing TFPI-2 (Hep3B-TFPI-2 group) and the Hep3B cells transfected with empty vector PCDNA3.1 (Hep3B-V group) or without transfection (Hep3B-P group) were subcutaneously transplanted into nude mice respectively to generate subcutaneous tumor xenografts. The volume of tumor xenograft was measured every three days, and the growth curve of tumor xenograft was drawn when the subcutaneous tumor xenograft was visible. The nude mice were killed three weeks after transplant, the volume of tumor xenograft was measured, and the total RNAs and proteins in tumor xenografts were extracted. The mRNA and protein expressions of TFPI-2 and VEGF (vascular endothelial growth factor) in tumor xenografts were analyzed by RFQ-PCR (real-time fluorescence quantitative PCR) and Western blotting, respectively. The expression of TFPI-2 protein and the MVD (microvessel density) in tumor xenografts were observed by immunohistochemistry. Results: The eventual tumor volume of tumor xenografts in Hep3B-TFPI-2 group was apparently smaller than those in Hep3B-V group and Hep3B-P group (both P < 0.05). The expression of mRNA and abundance of protein of TFPI-2 in Hep3B-TFPI-2 group were significantly higher than those in the other two groups (P < 0.05); while the expression of mRNA and abundance of protein of VEGF in Hep3B-TFPI-2 group were apparently lower than those in the other two groups. Compared with Hep3B-V group and Hep3B-P group, the inhibitory rates of VEGF protein expression in Hep3B-TFPI-2 group were 19.8% and 23.5%, respectively (P < 0.05). The MVD in Hep3B-TFPI-2 group was apparently lower than those in the other two groups (P < 0.05). Conclusion: TFPI-2 can significantly inhibit the growth and angiogenesis of subcutaneous tumor xenografts in nude mice established from hepatocarcinoma Hep3B cells. Copyright © 2013 by TUMOR.
ABSTRACT
Objective: To investigate the change in function of autophagy in hepatocarcinoma cell line HepG2 induced by Oxa (oxaliplatin), and to explore the role of autophagy in Oxa-induced cell death. Methods: The HepG2 cells were routinely cultured in vitro and treated either with different concentrations of Oxa alone or in combination with the autophagy inhibitor 3-MA (3-methyladenine ) or CQ (chloroquine). The cell viability was detected by CCK-8 (cell counting kit-8) assay. The formation of autophagosomes was observed directly under TEM (transmission electron microscope). Western-blotting was used to track the conversion of autophagic marker proteins LC3-I to LC3-II. Results: CCK-8 assay demonstrated that Oxa could dose-dependently induce the death of HepG2 cells, and when it was administrated in combination with 3-MA or CQ, Oxa could also significantly inhibit the growth of HepG2 cells. The formation of autophagosomes in HepG2 cells after Oxa treatment could be observed under TEM, as well as an increase in expression of LC3-II was found. The expression of LC3-II increased markedly after the inhibition of degradation pathway induced by CQ, which reflected an enhancement of autophagic flux induced by Oxa. 3-MA could inhibit the formation of autophagosomes in HepG2 cells. Conclusion: Oxa can enhance the function of autophagy in HepG2 cells and induce an increase in autophagosome formation. The autophagy inhibitors 3-MA and CQ can both facilitate the proliferative inhibition of hepatocarcinoma cell line HepG2 induced by Oxa, and they can also increase the antitumor effect of Oxa. Copyright © 2013 by TUMOR.
ABSTRACT
Objective To investigate the feasibility of monitoring the tumor's hypoxic changes by 99Tcm-2-(2-methyl-5-nitro-1H-imidazol-1-yl) ethyl eihydrogen phosphate (MNLS) imaging after radiotherapy.Methods (1) H22 cells were cultured and mice model with liver cancer xenografte was made.The mice were imaged at 0.5,1,2,3,4,6 and 8 h (six mice in each group) after injected with 7.4 MBq 99TcmMNLS when the tumor size reach about 1 cm.Then the mice were sacrificed.The T/NT and ％ID/g of each time point was calculated.(2) The liver cancer bearing mice of radiotherapy group (25 Gy) and control group were imaged at 0,24,48 h,and then the technique of ROI was adopted to calculate the T/NT at each time point in the two groups.Immunohistochemical stain method was used to evaluate the expression level of HIF-1α in liver cancer.(3) One-way analysis of variance,the least significant difference t test,two-sample t test and Spearman correlation analysis were performed.Results (1) The uptake of 99TcmMNLS in the liver cancer bearing mice was significant at 2 h after injection and the ％ID/g was the highest.99Tcm-MNLS was excreted mainly through kidneys.(2) The T/NT and HIF-1α expression level in radiotherapy group at 24 h (2.65±0.27,(50.62±3.78)％) were lower than those at the instant (3.35±0.19,(85.32±0.94)％,t=5.640,6.701,both P＜0.05),but higher than those at 48 h (2.23±0.52,(21.69±0.75)％,t=7.674,4.911,both P＜0.05).The T/NT and HIF-1α expression in the radiotherapy group were significantly higher than those in the control group at the instant (2.74 ± 0.29,(28.26 ± 1.70) ％,t =4.235,3.473,both P＜0.05) but lower at 48 h (3.15±0.88,(67.64±3.55) ％,t =7.902,3.258,both P＜0.05).However,no significant difference was observed at 24 h between radiotherapy group and the control group (2.98±0.16,(58.45±0.98) ％,t =0.525,2.043,both P＞0.05).The change of T/NT closely correlated with the expression of HIF-1α in both the radiotherapy group and control group(r,=0.793,0.756,both P＜0.05).Conclusion 99Tcm-MNLS hypoxia imaging has potential to monitor changes of hypoxia in tumor after radiotherapy.
ABSTRACT
Objective To study the biodistribution of 188Re-labeled stannic sulfur colloid in rabbit orthotopic VX2 liver cancer model by intratumoral injection and to evaluate its potential for endoradiotherapy.Methods 188Re-labeled stannic sulfur colloid was prepared with direct labeling method.The labeling efficiency and radiochemical purity were measured.Twelve rabbits xenografted by orthotopic VX2 liver cancer were used to determine the biodistribution of 188Re-labeled stannic sulfur colloid.Under CT guidance,37 MBq (0.1 ml) 188Relabeled stannic sulfur colloid was injected directly into the center of the tumor.Four rabbits were sacrificed after gamma imaging at 1,24,48 h post injection.The organ uptake was calculated as ％ID/g,the absorbed dose and T/NT ratio were calculated.One-way analysis of variance was used to analyze the data.Results The labeling efficiency of 188 Re-labeled stannic sulfur colloid was (98.23±0.25)％.The radiochemical purity was (94.23±0.54) ％ at 48 h.The radioactivity essentially accumulated in the tumor area and remained trapped up to 48 h.The radioactivity in other organs was at background level.The T/NT ratios were 88.22± 11.57,32.87±9.13 and 31.65± 10.11 at 1,24 and 48 h post injection respectively,with the corresponding tumor uptakes of (43.318±11.931) ％ID/g,(39.875±9.290) ％ID/g and (37.761±6.849) ％ID/g,which were much higher than those in normal tissues (F=77.350,97.577,417.072,all P＜0.01).Radiation dose to the tumor was (88.12 ± 12.21) Gy.Conclusions 188 Re-labeled stannic sulfur colloid may have a stable distribution at the site of orthotopic VX2 liver cancer after intratumoral injection.Thus it may have potential for the endoradiotherapy of liver cancer.
ABSTRACT
Objective To investigate whether HBx alone is sufficient to directly transform the nontransformed immortalized human liver cell line QSG7701 and induce hepatocellular carcinoma in vivo,and to explore preliminarily the pathogenic mechanism of transplantation tumor in nude mice.Methods The pCMVX/QSG7701 cells were vaccinated into subcutaneous tissue of nude mice.The pRcCMV2/QSG7701 and QSG7701 Cells were used as control.At the fifth weeks after vaccination,the nude mice were killed to observe whether a tumor was formed.The activity state and food intake of the nude mice was recorded.The texture,volume,and metastasis of transplantation tumor were observed grossly.The transplantation tumor was observed microscopically on the hematoxylin and eosin (HE) staining section.Immunohistochemical surfactant protein (SP) method was used to analyze the protein expression of mutant p53 and C-Myc genes.Results The transplantation tumor was occurred in all of the six nude mice vaccinated with pCMVX/QSG7701 cells at the second week after vaccination.No metastatic tumor was found in other organs.Transplanted tumor was not formed in all of the negative control groups.HE staining analysis confirmed that the character of transplanted tumor was hepatocellular carcinoma.p53 and C-Myc proteins were expressed in pCMVX/QSG7701,pRcCMV2/QSG7701,and QSG7701 cells,and their expression levels in the pCMVX/QSG7701 cells were significantly higher than those in the pRcCMV2/QSG7701 and QSG7701 cells,respectively(P ＜0.01).Conclusions HBx alone is sufficient to directly transform the non-transformed immortalized human liver cell line QSG7701 and induce hepatocellular carcinoma in vivo through up-regulating the expression of mutant p53 and C-Myc genes.
ABSTRACT
Objective: To detect the dynamic change of expression of CD133, a marker of liver cancer stem cell, at different stages of liver carcinogenesis induced by chemicals in C57BL/6J mice. Methods: The tumorigenesis and the growth of chemical (diethylnirtosamine/carbon tetrachloride/ethanol)-induced primary HCC (hepatocellular carcinoma) in 50 male C57BL/6J mice were observed. Another 45 male C57BL/6J mice which had not been exposed to carcinogenic chemicals were used as the normal controls. The mice were randomly sacrificed every two weeks, and the liver tissues were removed to observe the change of pathology. The expression of CD133 mRNA was detected by RFQ-PCR (real-time fluorescent quantitative PCR), and the expression of CD133 protein was detected by immunohistochemistry and Western blotting. The percentage of CD133-positive cells was analyzed by FCM (flow cytometry). Results: The primary HCC was successfully induced in male C57BL/6J mice after chemical intervention for 20 weeks. The results of RFQ-PCR and FCM showed that the expression level of CD133 in the chemicalinduced group was obviously higher than that in the normal control group after 8 weeks (P < 0.05, P < 0.001). The expression level of CD133 kept on rising in the chemical-induced group as time progressed, and it was significantly higher at the 16th week than at earlier stages (P < 0.001). Western blotting result showed that the CD133 weak expression could be observed at the 4th week. As time progressed, the expression level of CD133 protein was gradually increased. The result of immunohistochemistry showed that the expressions of CD133 in the chemical-induced group were weakly, moderately and strongly positive at the 8th, 16th and 20th week, respectively; while the expression of CD133 was negative in the normal control group. Conclusion: The liver cancer stem cell marker CD133 is involved in the development and progression of liver cancer, and its expression level is gradually increased in the process of carcinogenesis. Copyright © 2012 by TUMOR.
ABSTRACT
Objective To investigate dynamically characteristics of apparent diffusion coefficient (ADC) of MR diffusion-weighted imaging (DWI) in the rabbit VX-2 tumor model.Methods Forty New Zealand rabbits were included in the study and forty-seven rabbit VX-2 tumor models were raised by implanting directly and intrahepatically after abdominal cavity was opened.DWI was carried out periodically and respectively on seventh,fourteenth,and twenty-first day after implantation.Part samples of VX-2 tumors were studied by pathology.The distinction of VX-2 tumors on DWI was assessed by their ADC values.The statistical significance between different time groups,different area groups,or different b-value groups was calculated using SPSS12.0 software,respectively.Results ADC values of 47 VX-2 tumors in the area of tumor periphery,tumor center,and normal parenchyma around tumor were greater when b-value was 100 s/mm2 than those when b-value was 300 s/mm2 and the distinction of VX-2 tumor ADC in the area of tumor periphery,tumor center,and normal parenchyma around tumor between different b-value groups was significant,respectively( F =17.964,P ＜0.01 ; F =13.986,P ＜0.01 ; F =128.681,P ＜0.01 ).The ADC values in the area of normal liver parenchyma around tumor were greater than those in the area of VX-2 tumor periphery and tumor center when the b-value was 100 or 300 s/mm2.When b-value was the same( 100 or 300 s/mm2),the distinction of VX-2 tumor ADC between different areas was significant( F =176.586,P ＜0.01 ; F =55.089,P ＜0.01 ).The ADC of VX-2 tumor in the area of tumor periphery and tumor center became gradually low from seventh to fourteenth or twenty-first day after implantation and the distinction of ADC between different time groups but the area same (?) was significant( b =100 s/mm2,F =48.211,P ＜0.01 ;b =300 s/mm2,F =20.955,P ＜0.01 ).There were not obvious cellular necrosis in VX-2 tumors on seventh and fourteenth day after implantation but ADC of VX-2 tumor decreased unobviously because of cellular edemata in or around tumors.There were obvious cellular necrotic areas in VX-2 tumors on the twenty-first day after implantation.ADC of viable tumor cells in VX-2 tumors were lower on DWI than that in the area of normal liver parenchyma around tumor and ADC of dead tumor cells in VX-2 tumors were unequal,including high values,equal values,and low values but they were higher than that in the area of normal liver parenchyma around tumor after dead tumor cells had been liquified or had become cystic.Conclusions ADC is able to reflect objectively the diffusion of water molecules in the tumor and to reflect indirectly the degree of the growth and liquified necrosis of a tumor.ADC has an important and potential value in monitoring dynamical tumor growth and in evaluating malignant degree and therapeutic effect.
ABSTRACT
ObjectiveTo investigate its dynamic characteristics and pathological mechanism on magnatic resonance diffusion weighted imaging (DWI) after chemoembolization in rabbit liver VX-2 tumor model.MethodsForty New Zealand rabbits were included in the study and forty-seven rabbit VX-2 tumor models were raised by implanting directly and intrahepatically after abdominal cavity was opened.Forty VX-2 tumor models from them were divided into four groups.DWI was performed periodically and respectively for each group after chemoembolization.All VX-2 tumor samples of each group were studied by pathology.The distinction of VX-2 tumors on DWI was assessed by their apparent diffusion coefficient (ADC) values.The statistical significance between different time groups,different area groups,or different b-value groups was calculated using SPSS 12.0 software.ResultsWhen b-value was 100 s/mm2,ADC values in the area of VX-2 tumor periphery,VX-2 tumor central,or normal liver parenchyma around tumor became gradually low in sixteen hours after chemoembolization,and were the lowest at sixteenth hour,and then they increased gradually from sixteenth hour to fourty-eighth hour after chemoembolization.The distinction of ADC between different time groups was significant,respectively ( F =7.325,P ＜ 0.01 ; F =2.496,P ＜ 0.05 ; F =6.856,P ＜0.01 ).Cellular edema in the area of VX-2 tumor periphery or normal liver parenchyma around tumor increased quickly in sixteen hours after chemoembolization; however,from sixteenth hour to forty-eighth hour,cellular edema in the area of normal liver parenchyma around tumor decreased gradually and that in the area of VX-2 tumor periphery decreased lightly at first and then increased continually.Cellular necrosis in the area of VX-2 tumor periphery after chemoembolization was more significant than that before chemoembolization.The areas of dead cells in VX-2 tumors manifested low signal and high ADC value while the areas of viable cells manifested high signal and low ADC value.ConclusionsDWI is able to detect and discriminate tumor necrotic areas from viable cellular areas before and after chemoembolization.ADC of normal liver parenchyma and VX-2 tumor are influenced by intracellular edema,tissue cellular death,and microcirculation disturbance after chemoembolization.