AIM: To study the anti-hepatocarcinoma effects of 5-fluorouracil (5-Fu) encapsulated by galactosylceramide liposomes (5-Fu-GCL) in vivo and in vitro.METHODS: Tumor-bearing animal model and HepA cell line were respecti...AIM: To study the anti-hepatocarcinoma effects of 5-fluorouracil (5-Fu) encapsulated by galactosylceramide liposomes (5-Fu-GCL) in vivo and in vitro.METHODS: Tumor-bearing animal model and HepA cell line were respectively adopted to evaluate the anti-tumor effects of 5-Fu-GCL in vivoand in vitro. Tumor cell growth inhibition effects of 5-Fu-GCL in vitro were assessed by cell viability assay and MTT assay, In vivo experiment,the inhibitory effects on tumor growth were evaluated by tumor inhibition rate and animal survival days. High performance liquid chromatography was used to detect the concentration-time course of 5-Fu-GCL in intracellular fluid in vitro and the distribution of 5-Fu-GCL in liver tumor tissues in vivo. Apoptosis and cell cycle of tumor cells were demonstrated by flow cytometry.RESULTS: In vitro experiment, 5-Fu-GCL (6.25-100μmol/L) and free 5-Fu significantly inhibited HepA cell growth.Furthermore, IC50 of 5-Fu-GCL (34.5μmol/L) was lower than that of free 5-Fu (51.2μmol/L). In vivo experiment,5-Fu-GCL (20, 40, 80mg/kg) significantly suppressed the tumor growth in HepA bearing mice model. Compared with free 5-Fu, the area under curve of 5-Fu-GCL in intracellular fluid increased 2.6 times. Similarly, the distribution of 5-Fu-GCL in liver tumor tissues was significantly higher than that of free 5-Fu. After being treated with 5-Fu-GCL, the apoptotic rate and the proportion of HepA cells in the S phase increased, while the proportion in the G0/G1 and G2/M phases decreased.CONCLUSION: 5-Fu-GCL appears to have anti-hepato-carcinoma effects and its drug action is better than free 5-Fu. its mechanism is partly related to increased drug concentrations in intracallular fluid and liver tumor tissues,enhanced tumor cell apoptotic Fate and arrest of call cycle in S phase.展开更多
目的探讨骨髓中自然杀伤T细胞(nature killer T cell,NKT)在再生障碍性贫血免疫介导发病机制中的作用。方法选取14例确诊儿童再生障碍性贫血为再生障碍性贫血组,同年龄正常儿童10例为对照组。流式细胞术检测两组儿童骨髓中NKT含量;利用...目的探讨骨髓中自然杀伤T细胞(nature killer T cell,NKT)在再生障碍性贫血免疫介导发病机制中的作用。方法选取14例确诊儿童再生障碍性贫血为再生障碍性贫血组,同年龄正常儿童10例为对照组。流式细胞术检测两组儿童骨髓中NKT含量;利用免疫磁珠法分离纯化NKT细胞后,分别在4种不同体系下进行扩增培养。A:α-Galcer+rhIL-2;B:α-Galcer+rhIL-2+rhG-CSF;C:OCH+rhIL-2;D:OCH+rhIL-2+rhG-CSF。测定NKT细胞在不同培养条件下的扩增倍数;利用酶联免疫斑点技术(Elispot)测定NKT细胞扩增活化后表达IFN-γ、IL-4的斑点形成细胞数(SFCs)。结果再生障碍性贫血组骨髓中NKT百分率(0.827%±0.022%)明显低于对照组(1.033%±0.073%,Z=-3.810,P=0.000)。在各培养体系中,再生障碍性贫血组和对照组骨髓NKT均可明显扩增。在含有α-Galcer培养体系A和B中,再生障碍性贫血组及对照组NKT细胞的扩增能力均高于OCH体系C和D,差异有统计学意义(P<0.01或P<0.05)。在培养体系中加入rhG-CSF后,再生障碍性贫血组骨髓NKT细胞的扩增能力下降(P<0.01),但表达IFN-γ的SFCs明显减少(P均<0.01);而表达IL-4的SFCs明显升高(P均<0.01)。结论再生障碍性贫血患儿骨髓NKT的数量和功能均明显低于正常儿童。但再生障碍性贫血骨髓在接受细胞配体(OCH或α-Galcer)+rhIL-2+rhG-CSF作用后,在获得NKT细胞一定程度扩增的同时,提高NKT的IL-4表达,降低NKT的IFN-γ表达,抑制T细胞向Th1分化,而促进T细胞向Th2分化,逆转再生障碍性贫血Th1/Th2异常失衡的免疫介导致病机制,可能成为获得性再生障碍性贫血的有效治疗途径。展开更多
Objective:The aim of this study was to investigate the inhibition effect of natural killer T(NKT) cells on transplantation hepatocellular carcinoma in mice.Methods:α-galactosylceramide(α-GalCer)-pulsed DC and Hep S ...Objective:The aim of this study was to investigate the inhibition effect of natural killer T(NKT) cells on transplantation hepatocellular carcinoma in mice.Methods:α-galactosylceramide(α-GalCer)-pulsed DC and Hep S were prepared as stimulus.Hepatoma xenograft model was established and mice were randomly divided into 4 groups(n=13 each group):(1) control group,intravenous injection of the same volume of saline.(2) mature DC group,intravenous injection of mature DC cells(4×106 cells).(3) α-GalCer-pulsed HepS group,intravenous injection of α-GalCer-pulsed HepS(4×106 cells).(4) α-GalCer-pulsed mature DC group,intravenous injection of α-GalCer-pulsed DC(4×106 cells).The changes of tumor volume in mice and survival period were measured every 2 days.Percentage of NKT cells in spleens and cytotoxicity of spleen cells were detected by flow cytometry.Tumor tissues were analyzed by histopathological examination.Results:In α-GalCer-pulsed Heps and DC groups,the average survival period was prolonged and tumor volume was markedly decreased,spleen cells and NKT cells were significantly increased,and tumor necrosis was evident,compared to the control group.Conclusion:α-GalCer-pulsed DC and HepS could activate NKT cells in vivo,also increase NKT cells cytotoxicity,inhibit the growth of hepatomas and prolong survival period.展开更多
基金Supported by the Key Teacher Foundation of Ministry of Education of China, No. 1869 Young Teacher Foundation of Department of Education of Anhui Province, No. 2000jp112
文摘AIM: To study the anti-hepatocarcinoma effects of 5-fluorouracil (5-Fu) encapsulated by galactosylceramide liposomes (5-Fu-GCL) in vivo and in vitro.METHODS: Tumor-bearing animal model and HepA cell line were respectively adopted to evaluate the anti-tumor effects of 5-Fu-GCL in vivoand in vitro. Tumor cell growth inhibition effects of 5-Fu-GCL in vitro were assessed by cell viability assay and MTT assay, In vivo experiment,the inhibitory effects on tumor growth were evaluated by tumor inhibition rate and animal survival days. High performance liquid chromatography was used to detect the concentration-time course of 5-Fu-GCL in intracellular fluid in vitro and the distribution of 5-Fu-GCL in liver tumor tissues in vivo. Apoptosis and cell cycle of tumor cells were demonstrated by flow cytometry.RESULTS: In vitro experiment, 5-Fu-GCL (6.25-100μmol/L) and free 5-Fu significantly inhibited HepA cell growth.Furthermore, IC50 of 5-Fu-GCL (34.5μmol/L) was lower than that of free 5-Fu (51.2μmol/L). In vivo experiment,5-Fu-GCL (20, 40, 80mg/kg) significantly suppressed the tumor growth in HepA bearing mice model. Compared with free 5-Fu, the area under curve of 5-Fu-GCL in intracellular fluid increased 2.6 times. Similarly, the distribution of 5-Fu-GCL in liver tumor tissues was significantly higher than that of free 5-Fu. After being treated with 5-Fu-GCL, the apoptotic rate and the proportion of HepA cells in the S phase increased, while the proportion in the G0/G1 and G2/M phases decreased.CONCLUSION: 5-Fu-GCL appears to have anti-hepato-carcinoma effects and its drug action is better than free 5-Fu. its mechanism is partly related to increased drug concentrations in intracallular fluid and liver tumor tissues,enhanced tumor cell apoptotic Fate and arrest of call cycle in S phase.
文摘目的探讨骨髓中自然杀伤T细胞(nature killer T cell,NKT)在再生障碍性贫血免疫介导发病机制中的作用。方法选取14例确诊儿童再生障碍性贫血为再生障碍性贫血组,同年龄正常儿童10例为对照组。流式细胞术检测两组儿童骨髓中NKT含量;利用免疫磁珠法分离纯化NKT细胞后,分别在4种不同体系下进行扩增培养。A:α-Galcer+rhIL-2;B:α-Galcer+rhIL-2+rhG-CSF;C:OCH+rhIL-2;D:OCH+rhIL-2+rhG-CSF。测定NKT细胞在不同培养条件下的扩增倍数;利用酶联免疫斑点技术(Elispot)测定NKT细胞扩增活化后表达IFN-γ、IL-4的斑点形成细胞数(SFCs)。结果再生障碍性贫血组骨髓中NKT百分率(0.827%±0.022%)明显低于对照组(1.033%±0.073%,Z=-3.810,P=0.000)。在各培养体系中,再生障碍性贫血组和对照组骨髓NKT均可明显扩增。在含有α-Galcer培养体系A和B中,再生障碍性贫血组及对照组NKT细胞的扩增能力均高于OCH体系C和D,差异有统计学意义(P<0.01或P<0.05)。在培养体系中加入rhG-CSF后,再生障碍性贫血组骨髓NKT细胞的扩增能力下降(P<0.01),但表达IFN-γ的SFCs明显减少(P均<0.01);而表达IL-4的SFCs明显升高(P均<0.01)。结论再生障碍性贫血患儿骨髓NKT的数量和功能均明显低于正常儿童。但再生障碍性贫血骨髓在接受细胞配体(OCH或α-Galcer)+rhIL-2+rhG-CSF作用后,在获得NKT细胞一定程度扩增的同时,提高NKT的IL-4表达,降低NKT的IFN-γ表达,抑制T细胞向Th1分化,而促进T细胞向Th2分化,逆转再生障碍性贫血Th1/Th2异常失衡的免疫介导致病机制,可能成为获得性再生障碍性贫血的有效治疗途径。
基金National Natural Science Foundation of China (Grant No.90713010)"863"Program from the Ministry of Science and Technology of China (Grant No.2006AA09Z405).
基金Supported by a grant of Eleventh Five Year Plan project of Nanjing Military Region (No. 06MA45)
文摘Objective:The aim of this study was to investigate the inhibition effect of natural killer T(NKT) cells on transplantation hepatocellular carcinoma in mice.Methods:α-galactosylceramide(α-GalCer)-pulsed DC and Hep S were prepared as stimulus.Hepatoma xenograft model was established and mice were randomly divided into 4 groups(n=13 each group):(1) control group,intravenous injection of the same volume of saline.(2) mature DC group,intravenous injection of mature DC cells(4×106 cells).(3) α-GalCer-pulsed HepS group,intravenous injection of α-GalCer-pulsed HepS(4×106 cells).(4) α-GalCer-pulsed mature DC group,intravenous injection of α-GalCer-pulsed DC(4×106 cells).The changes of tumor volume in mice and survival period were measured every 2 days.Percentage of NKT cells in spleens and cytotoxicity of spleen cells were detected by flow cytometry.Tumor tissues were analyzed by histopathological examination.Results:In α-GalCer-pulsed Heps and DC groups,the average survival period was prolonged and tumor volume was markedly decreased,spleen cells and NKT cells were significantly increased,and tumor necrosis was evident,compared to the control group.Conclusion:α-GalCer-pulsed DC and HepS could activate NKT cells in vivo,also increase NKT cells cytotoxicity,inhibit the growth of hepatomas and prolong survival period.
