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Volume 7,Issue 1,2000 Table of Contents
2000, 7(1):1-2.
DOI: 10.3872/j.issn.1007-385X.2000.1.001
Abstract:
Objective: To study the cellular immune response and the anti-tumor effects induced by intramuscular DNA immu- nization with HER2/neu extracellular domain gene. Methods: HEH2/neu oncogene extracelltilar domain gene was isolated and inserted to pCDNA3 expression vector. The cellular immune response and its anti-tumor effects were analyzed after intramuscular DNA immunization. Results: A specific cytotoxic activity by spleen cells from HER2/neu ECD gene immunized mice was found in vitro. Tumor growth was inhibited after tumor challenge in vivo in immunized mice. Conclusion: Specific cellular immune re- sponse and anti-tumor effect were elicited by HEB2/neu extracellular domain gene immunization.
Objective: To study the cellular immune response and the anti-tumor effects induced by intramuscular DNA immu- nization with HER2/neu extracellular domain gene. Methods: HEH2/neu oncogene extracelltilar domain gene was isolated and inserted to pCDNA3 expression vector. The cellular immune response and its anti-tumor effects were analyzed after intramuscular DNA immunization. Results: A specific cytotoxic activity by spleen cells from HER2/neu ECD gene immunized mice was found in vitro. Tumor growth was inhibited after tumor challenge in vivo in immunized mice. Conclusion: Specific cellular immune re- sponse and anti-tumor effect were elicited by HEB2/neu extracellular domain gene immunization.
2000, 7(1):3-5.
DOI: 10.3872/j.issn.1007-385X.2000.1.002
Abstract:
To explore the mechanism of prevention and therapy by retinoic acid in cancer and isolate ATRA-target gene and explore the mechanism of expression regulation of mitochondrial gene. Methods： AP-PCR, hybridization, sequencing etc., were used. Results ： ATRA up-regulated the expression of mitochondrion gene ATPase subunit 6, the change lasted in whole period of ATRA treatment (from first day to seventh day). The up-regulation was confirmed by Northen blot analysis. Two possible retinoic acid responsive elements (RAREs) were located in the regulation region of mitochondrion genome. Conclusion： The mitochondrion gene expression by ATRA might be regulated through the interaction of retinoic acid receptor (RAR) with RARE in mitochondrion genome
To explore the mechanism of prevention and therapy by retinoic acid in cancer and isolate ATRA-target gene and explore the mechanism of expression regulation of mitochondrial gene. Methods： AP-PCR, hybridization, sequencing etc., were used. Results ： ATRA up-regulated the expression of mitochondrion gene ATPase subunit 6, the change lasted in whole period of ATRA treatment (from first day to seventh day). The up-regulation was confirmed by Northen blot analysis. Two possible retinoic acid responsive elements (RAREs) were located in the regulation region of mitochondrion genome. Conclusion： The mitochondrion gene expression by ATRA might be regulated through the interaction of retinoic acid receptor (RAR) with RARE in mitochondrion genome
2000, 7(1):6-10.
DOI: 10.3872/j.issn.1007-385X.2000.1.003
Abstract:
Purpose to investigate the different in vitrofunction of targetable non-viral vector containing poly-L-lysine or protamine. Methods： Using GV1 and GV2 targetable non-viral vectors, the influences of the poly-L-lysine and protamine on in vitro gene transfer efficiency and the course of gene expression were observed. Results： β-galactosidase was expressed at intermediate level (50%) in A375 cells using a complex containing either protamine or poly-L-lysine. However, in case of ABAE cells, β-galactosidase expression level was low (20%) transferred with a complex containing protamine. On the contrary, β-galactosidase expression was at high level (70%) provided that protamine was replaced with poly-L-lysine. In addition, β-galactosidase activity reached the peak at the 6th day after transfection with the complex containing protamine. The expression was not altered with subsequent subcultures, at least for 3 passages. Using poly-L-lysine, the expression peak in A375 reached the peak at the 7th day after transfection, but the level declined along with subsequent passages of cells. Conclusion: The apllication of protamine in VEGF receptor mediated gene delivery system was limited.
Purpose to investigate the different in vitrofunction of targetable non-viral vector containing poly-L-lysine or protamine. Methods： Using GV1 and GV2 targetable non-viral vectors, the influences of the poly-L-lysine and protamine on in vitro gene transfer efficiency and the course of gene expression were observed. Results： β-galactosidase was expressed at intermediate level (50%) in A375 cells using a complex containing either protamine or poly-L-lysine. However, in case of ABAE cells, β-galactosidase expression level was low (20%) transferred with a complex containing protamine. On the contrary, β-galactosidase expression was at high level (70%) provided that protamine was replaced with poly-L-lysine. In addition, β-galactosidase activity reached the peak at the 6th day after transfection with the complex containing protamine. The expression was not altered with subsequent subcultures, at least for 3 passages. Using poly-L-lysine, the expression peak in A375 reached the peak at the 7th day after transfection, but the level declined along with subsequent passages of cells. Conclusion: The apllication of protamine in VEGF receptor mediated gene delivery system was limited.
2000, 7(1):11-14.
DOI: 10.3872/j.issn.1007-385X.2000.1.004
Abstract:
To investigate the effects of novel targeted non-viral vector in gene therapy of human glioma. Methods: The EGF-R targeting gene delivery system GE7 was constructed. Human glioma cell line U251 was transfected in vitro with β-gal as reportor gene and p21 as therapeutic gene using this gene delivery system. By means of the assay of β-galactosidase staining, Western blotting, in situ end labeling apoptosis cells and DNA ladder, the transferring of exogenous genes and the apoptosis of the tumor cells were examined. Results: It was showed that gene transfer efficiency is over 80%. When transfected with p21 gene, the growth of cells was inhibited significantly, and the apoptosis was detected in the transfected cell by the methods of in situ end labeling and DNA ladder. Conclusion: The GE7 gene delivery system has the ability to transfer exogenous gene to tumor cells and the expression of the therapeutic gene can inhibit the growth of the cells.
To investigate the effects of novel targeted non-viral vector in gene therapy of human glioma. Methods: The EGF-R targeting gene delivery system GE7 was constructed. Human glioma cell line U251 was transfected in vitro with β-gal as reportor gene and p21 as therapeutic gene using this gene delivery system. By means of the assay of β-galactosidase staining, Western blotting, in situ end labeling apoptosis cells and DNA ladder, the transferring of exogenous genes and the apoptosis of the tumor cells were examined. Results: It was showed that gene transfer efficiency is over 80%. When transfected with p21 gene, the growth of cells was inhibited significantly, and the apoptosis was detected in the transfected cell by the methods of in situ end labeling and DNA ladder. Conclusion: The GE7 gene delivery system has the ability to transfer exogenous gene to tumor cells and the expression of the therapeutic gene can inhibit the growth of the cells.
2000, 7(1):15-17.
DOI: 10.3872/j.issn.1007-385X.2000.1.005
Abstract:
To induce an immune response by intramuscular immunization with HER2/neu oncogene extracellular domain (ECD) DNA and observe the effect in vivo by an abortion model in mice. Methods: Human and rat HER2/neu ECD gene were cloned and inserted into expressive vector pCDNA3. The DNA immunization, expression of HER2/neu ECD, antibody detection and abortion in immunized mice were studied. Results: HER2/neu ECD protein expressed on muscle cells in the injected site, specific antibody to HER2/neu was induced by DNA immunization. The average of the offspring per delivery from immunized mice was significantly lower than that from control group, and the abortive embryos were found in uterus from immunized mice. Conclusion: The results show that both human and rat HER2/neu ECD gene can induced effective immune response by DNA immunization.
To induce an immune response by intramuscular immunization with HER2/neu oncogene extracellular domain (ECD) DNA and observe the effect in vivo by an abortion model in mice. Methods: Human and rat HER2/neu ECD gene were cloned and inserted into expressive vector pCDNA3. The DNA immunization, expression of HER2/neu ECD, antibody detection and abortion in immunized mice were studied. Results: HER2/neu ECD protein expressed on muscle cells in the injected site, specific antibody to HER2/neu was induced by DNA immunization. The average of the offspring per delivery from immunized mice was significantly lower than that from control group, and the abortive embryos were found in uterus from immunized mice. Conclusion: The results show that both human and rat HER2/neu ECD gene can induced effective immune response by DNA immunization.
2000, 7(1):18-20.
DOI: 10.3872/j.issn.1007-385X.2000.1.006
Abstract:
To study the cellular immune response and the anti-tumor effects induced by intramuscular DNA immunization with HER2/neu extracellular domain gene. Methods: HER2/neu oncogene extracellular domain gene was isolated and inserted to pCDNA3 expression vector. The cellular immune response and its anti-tumor effects were analyzed after intramuscular DNA immunization. Results: A specific cytotoxic activity by spleen cells from HER2/neu ECD gene immunized mice was found in vitro. Tumor growth was inhibited after tumor challenge in vivo in immunized mice. Conclusion: Specific cellular immune response and anti-tumor effect were elicited by HER2/neu extracellular domain gene immunization.
To study the cellular immune response and the anti-tumor effects induced by intramuscular DNA immunization with HER2/neu extracellular domain gene. Methods: HER2/neu oncogene extracellular domain gene was isolated and inserted to pCDNA3 expression vector. The cellular immune response and its anti-tumor effects were analyzed after intramuscular DNA immunization. Results: A specific cytotoxic activity by spleen cells from HER2/neu ECD gene immunized mice was found in vitro. Tumor growth was inhibited after tumor challenge in vivo in immunized mice. Conclusion: Specific cellular immune response and anti-tumor effect were elicited by HER2/neu extracellular domain gene immunization.
2000, 7(1):21-24.
DOI: 10.3872/j.issn.1007-385X.2000.1.007
Abstract:
To investigate the effect of IL-2 gene modification on expression of cytokines and chemokines in dendritic cells (DCs) and IL-2 expression in mouse organs afterin vivoDC immunization and to determine the improvement of microenvironment of antigen presentation and its potential mechanism. Methods: The IL-2 levels in DC culture, mouse serum and organs after IL-2 gene-modified DC(DC-IL-2) injection in vivowere examined by ELISA. The mRNA expressions of cytokine and chemokines (IL-2, IFN-γ, IL-12, GM-CSF, IL-4, IL-10, TNF-α, IL-1β, MIP1β, MCP-1, RANTES and IP10) in DC-IL-2 were analyzed by RT-PCR. The expression of IL-2Rα on surface of DC-IL-2 was analyzed by FACS. Results: After IL-2 gene modification, DCs could secret high level of IL-2, IFN-γ and IL-12. The expression of IL-2Rα on the surface of DC-IL-2 was increased. mRNAs of various cytokines and chemokines were detected. The levels of IL-2 increased immediately in mouse serum, lung, spleen and liver after DC-IL-2 injection in vivo. Conclusion: The IL-2 gene modification could enhance the biologic activity of the DCs and the microenvironment of antigen presentation was optimized.
To investigate the effect of IL-2 gene modification on expression of cytokines and chemokines in dendritic cells (DCs) and IL-2 expression in mouse organs afterin vivoDC immunization and to determine the improvement of microenvironment of antigen presentation and its potential mechanism. Methods: The IL-2 levels in DC culture, mouse serum and organs after IL-2 gene-modified DC(DC-IL-2) injection in vivowere examined by ELISA. The mRNA expressions of cytokine and chemokines (IL-2, IFN-γ, IL-12, GM-CSF, IL-4, IL-10, TNF-α, IL-1β, MIP1β, MCP-1, RANTES and IP10) in DC-IL-2 were analyzed by RT-PCR. The expression of IL-2Rα on surface of DC-IL-2 was analyzed by FACS. Results: After IL-2 gene modification, DCs could secret high level of IL-2, IFN-γ and IL-12. The expression of IL-2Rα on the surface of DC-IL-2 was increased. mRNAs of various cytokines and chemokines were detected. The levels of IL-2 increased immediately in mouse serum, lung, spleen and liver after DC-IL-2 injection in vivo. Conclusion: The IL-2 gene modification could enhance the biologic activity of the DCs and the microenvironment of antigen presentation was optimized.
2000, 7(1):25-27.
DOI: 10.3872/j.issn.1007-385X.2000.1.008
Abstract:
To explore the adjustment of whole peptidoglycans of bifidobacteria to macrophage functions. Methods： The level of interleukin 1， tumor necrosis factor α and nitric oxide (NO) produced by peritoneal macrophages of nude mice was detected by employing mouse thymocyte proliferation method killing L929 cell method in vitro and Griess reagent,after whole peptidoglycans were injected into the nude mice peritoneally. Simultaneously, tumoricidal activity of themacrophages in vitro was observed. Results： The content of interleukin 1， tumor necrosis factor α and nitric oxide (NO) secreted from peritoneal macrophages of nude mice was significantly higher in the whole peptidoglycan group when compared with the control group. Futhermore, the tumoricidal activity of the macrophages in vitro was markedly promoted (P<0.01). Conclusion： Whole peptidoglycans of bifidobacteria bifidum could activated macrophages to secrete a lot of cytotoxicity effectors,and elevate its tumoricidal activity.
To explore the adjustment of whole peptidoglycans of bifidobacteria to macrophage functions. Methods： The level of interleukin 1， tumor necrosis factor α and nitric oxide (NO) produced by peritoneal macrophages of nude mice was detected by employing mouse thymocyte proliferation method killing L929 cell method in vitro and Griess reagent,after whole peptidoglycans were injected into the nude mice peritoneally. Simultaneously, tumoricidal activity of themacrophages in vitro was observed. Results： The content of interleukin 1， tumor necrosis factor α and nitric oxide (NO) secreted from peritoneal macrophages of nude mice was significantly higher in the whole peptidoglycan group when compared with the control group. Futhermore, the tumoricidal activity of the macrophages in vitro was markedly promoted (P<0.01). Conclusion： Whole peptidoglycans of bifidobacteria bifidum could activated macrophages to secrete a lot of cytotoxicity effectors,and elevate its tumoricidal activity.
2000, 7(1):28-31.
DOI: 10.3872/j.issn.1007-385X.2000.1.009
Abstract:
To study the main features of CH50, a recombinant polypeptide of human fibronectin, to activate macrophages in vivo and its anti-tumor function. Methods： CH50 was injected and IFN-γ gene was transfected in mice several kinds of factors produced by macrophages were determined. The growth of tumor in mice was also measured. Results： CH50 could enhance the production of such factors as NO, TNF and IL-1 by macrophages, but the activation of macrophages was relatively slow when CH50 was used in vivo alone. CH50 and IFN-γ could synergistically activate macrophages rapidly in vivo no matter whether the injection of CH50 or the transfection of IFN-γ gene was performed first. Injection of CH50 alone inhibited the formation of tumor nodes in a dose-dependent manner. There was strong inhibition of low dosage of CH50 on the formation of tumor nodes smaller than 1 mm, and high dosage of CH50 on those bigger than 1 mm. A stronger inhibition on the growth of tumor in vivo was obtained by the synergistic effect of CH50 and IFN-γ. Conclusion： CH50 and IFN-γ, as double-signal factors for activation of macrophages, will be potentially useful in tumor therapy.
To study the main features of CH50, a recombinant polypeptide of human fibronectin, to activate macrophages in vivo and its anti-tumor function. Methods： CH50 was injected and IFN-γ gene was transfected in mice several kinds of factors produced by macrophages were determined. The growth of tumor in mice was also measured. Results： CH50 could enhance the production of such factors as NO, TNF and IL-1 by macrophages, but the activation of macrophages was relatively slow when CH50 was used in vivo alone. CH50 and IFN-γ could synergistically activate macrophages rapidly in vivo no matter whether the injection of CH50 or the transfection of IFN-γ gene was performed first. Injection of CH50 alone inhibited the formation of tumor nodes in a dose-dependent manner. There was strong inhibition of low dosage of CH50 on the formation of tumor nodes smaller than 1 mm, and high dosage of CH50 on those bigger than 1 mm. A stronger inhibition on the growth of tumor in vivo was obtained by the synergistic effect of CH50 and IFN-γ. Conclusion： CH50 and IFN-γ, as double-signal factors for activation of macrophages, will be potentially useful in tumor therapy.
2000, 7(1):32-34.
DOI: 10.3872/j.issn.1007-385X.2000.1.010
Abstract:
To investigate the effect of recombinant BCG secreting IL-2 (rBCG-IL-2) on peritoneal macrophages of mouse and inquire into the effect of rBCG-IL-2 on immunity of host. Methods: Gene engineering technology, electricity transference and inoculation tumor cell are adopted; MTT assay is used to determine tumoricidal effect of macrophages. Results: Tumoricidal effect of macrophages of rBCG-IL-2 groups was higher than those of the control groups and BCG groups, and the tumoricidal effect of macrophages could reach a high state in a week. Conclusion: rBCG-IL-2 can raise tumoricidal effect of macrophages, improve immunity of host and is a prospect biological preparation.
To investigate the effect of recombinant BCG secreting IL-2 (rBCG-IL-2) on peritoneal macrophages of mouse and inquire into the effect of rBCG-IL-2 on immunity of host. Methods: Gene engineering technology, electricity transference and inoculation tumor cell are adopted; MTT assay is used to determine tumoricidal effect of macrophages. Results: Tumoricidal effect of macrophages of rBCG-IL-2 groups was higher than those of the control groups and BCG groups, and the tumoricidal effect of macrophages could reach a high state in a week. Conclusion: rBCG-IL-2 can raise tumoricidal effect of macrophages, improve immunity of host and is a prospect biological preparation.
11 Multiple Low-Dose RIT with 131 I-C50 in Controlling and Preventing Metastasis in Tumor-Bearing Mice
Ding Yong
,
Tian Jiahe
,
Zhang Jinming
,
Cao Liming
,
Wang Zheng
,
Chen Yingmao
,
Zhang Shuwen
,
Guan Zhiwei
,
Feng Huiru
2000, 7(1):39-41.
DOI: 10.3872/j.issn.1007-385X.2000.1.012
Abstract:
To study the efficiency of radio-immuno-therapy (RIT) with 131 I-C50 to precaution and treatment of tumor metastasis and develop a new metastasis suppression method. Methods: The effect was judged through body weight loss, tumor burden, life-span, and metastatic. Compared among the multiple low-dose RIT, chemotherapy, surgical resection and controls in different administration time after implantation of tumor and resection of tumor. Results: When the treatment was initiated at 7th day after implantation, the tumor suppression effects of treatment groups were significantly better than those of controls, with the best one in multiple low-dose RIT. Better results were achieved by starting treatment earlier. Surgical resection at 15th day after implantation, multiple low-dose RIT groups gained longer life-span and less metastatic nodules than others. Early operation improved therapeutic efficacy, especially in multiple low-dose RIT. Conclusion: Multiple low-dose RIT alone is better than chemotherapy and surgical treating. Early doing RIT is better than latter. One low-dose RIT plus with operation had even better tumor suppression effect than RIT alone. Low-dose RIT co-operated with operation as early as possible can enhance the effects of treatment.
To study the efficiency of radio-immuno-therapy (RIT) with 131 I-C50 to precaution and treatment of tumor metastasis and develop a new metastasis suppression method. Methods: The effect was judged through body weight loss, tumor burden, life-span, and metastatic. Compared among the multiple low-dose RIT, chemotherapy, surgical resection and controls in different administration time after implantation of tumor and resection of tumor. Results: When the treatment was initiated at 7th day after implantation, the tumor suppression effects of treatment groups were significantly better than those of controls, with the best one in multiple low-dose RIT. Better results were achieved by starting treatment earlier. Surgical resection at 15th day after implantation, multiple low-dose RIT groups gained longer life-span and less metastatic nodules than others. Early operation improved therapeutic efficacy, especially in multiple low-dose RIT. Conclusion: Multiple low-dose RIT alone is better than chemotherapy and surgical treating. Early doing RIT is better than latter. One low-dose RIT plus with operation had even better tumor suppression effect than RIT alone. Low-dose RIT co-operated with operation as early as possible can enhance the effects of treatment.
2000, 7(1):42-45.
DOI: 10.3872/j.issn.1007-385X.2000.1.013
Abstract:
To investigate the variation in vitro bystander effect among human lung cancer cell lines after transfection of pCEAcd-tk. Methods: pCEAcd-tk was extracted and transfected to six human lung cancer cell lines. Their sensitivity to 5-fluorocytosine (5-FC) and ganciclovir (GCV) was measured by MTT assay. Their bystander effect was also determined. Results: 1. The efficiency of lipid mediated gene transfer was very low; 2. Two out of six human lung cancer cell lines(SPCA-1，A549) have a high sensitivity to 5-FC and GCV; 3.Bystander effect: Two out of six human cancer cell lines（SPCA-1，SC） have a good bystander effect, and one out of them（A49）has a intermediate bystander effect, but three out of them （GLC-82，HLAMP and L-78）have a poor bystander effect. Conclusion： There are a variation in vitro bystander effect in six human lung cancer cell lines after transfected with pCEAcd-tk.
To investigate the variation in vitro bystander effect among human lung cancer cell lines after transfection of pCEAcd-tk. Methods: pCEAcd-tk was extracted and transfected to six human lung cancer cell lines. Their sensitivity to 5-fluorocytosine (5-FC) and ganciclovir (GCV) was measured by MTT assay. Their bystander effect was also determined. Results: 1. The efficiency of lipid mediated gene transfer was very low; 2. Two out of six human lung cancer cell lines(SPCA-1，A549) have a high sensitivity to 5-FC and GCV; 3.Bystander effect: Two out of six human cancer cell lines（SPCA-1，SC） have a good bystander effect, and one out of them（A49）has a intermediate bystander effect, but three out of them （GLC-82，HLAMP and L-78）have a poor bystander effect. Conclusion： There are a variation in vitro bystander effect in six human lung cancer cell lines after transfected with pCEAcd-tk.
2000, 7(1):46-49.
DOI: 10.3872/j.issn.1007-385X.2000.1.014
Abstract:
To study the vaccine potency of gene-modified tumor cells, we have constructed IL-12,B7-1 and GM-CSF express vector using retrovirus. Methods: It was transfected into EL-4 thymic lymphoma cells respectively and the effect of gene transduction on anti-tumor immunity were investigated. Results: The tumorigenicity of EL-4/IL-12 transfectant in C57BL/6 synergistical mice was decreased significantly after implanted with EL-4/IL-12 transfectant compaired with EL-4/Wt or EL-4/Neo groups (P＜001).The systemic protective immunity was induced against the challenge with EL-4/Wt (in 10/15 mice) after the rejection of EL-4/IL-12 in vivo experiment,a stronger CTL activity against EL-4/Wt cells and a weak killer activity against syngeneic Lewis Lung Carcinoma cells were obtained in ５１Cr release assay. In vivo depletion analysis suggested that the decrcased tumorigenicity mainly depended on CD4＋,CD8＋ and NK cells. Therapeutic vaccines with EL-4/IL-12 cells could retard the growth to estabished EL-4/Wt tumors significantly compared with those of EL-4/Neo (P＜0005)， combination of therapeutic vaccines of EL-4/IL-12 and EL-4/B7-1 result in the enhanced the therapeutic effect of each single transficants (P＜0005) in this experimental model. Conclution: These studies suggested that immunogene treatment using IL-12 is effective in hematopoietic malignancy,and combination of IL-12 with B7-1 have a aplication value in human cancer treatment in the near future.
To study the vaccine potency of gene-modified tumor cells, we have constructed IL-12,B7-1 and GM-CSF express vector using retrovirus. Methods: It was transfected into EL-4 thymic lymphoma cells respectively and the effect of gene transduction on anti-tumor immunity were investigated. Results: The tumorigenicity of EL-4/IL-12 transfectant in C57BL/6 synergistical mice was decreased significantly after implanted with EL-4/IL-12 transfectant compaired with EL-4/Wt or EL-4/Neo groups (P＜001).The systemic protective immunity was induced against the challenge with EL-4/Wt (in 10/15 mice) after the rejection of EL-4/IL-12 in vivo experiment,a stronger CTL activity against EL-4/Wt cells and a weak killer activity against syngeneic Lewis Lung Carcinoma cells were obtained in ５１Cr release assay. In vivo depletion analysis suggested that the decrcased tumorigenicity mainly depended on CD4＋,CD8＋ and NK cells. Therapeutic vaccines with EL-4/IL-12 cells could retard the growth to estabished EL-4/Wt tumors significantly compared with those of EL-4/Neo (P＜0005)， combination of therapeutic vaccines of EL-4/IL-12 and EL-4/B7-1 result in the enhanced the therapeutic effect of each single transficants (P＜0005) in this experimental model. Conclution: These studies suggested that immunogene treatment using IL-12 is effective in hematopoietic malignancy,and combination of IL-12 with B7-1 have a aplication value in human cancer treatment in the near future.
2000, 7(1):50-52.
DOI: 10.3872/j.issn.1007-385X.2000.1.015
Abstract:
To study the role of TGF-β1 and its type Ⅱ receptor in pathogenesis of hepatocellular carcinoma. Methods: The TGF-β1 and its type Ⅱ receptor in the tumor and surrounding tissue in 30 cases with hepatocellular carcinoma were investigated. Results: Levels of protein and mRNA were detected by immuno-histochemistry, hybridization and semi-quantitative analysis. The results showed that TGF-β1 in the tumor and surrounding tissue was overexpressed, and the levels of latter was even higher. TGF-β1 type Ⅱ receptor expressed in surrounding tissue, but it could not be found in most tumor tissues. Conclusion: The lack of TGF-β receptor in the tumor cell may be the way of cancer cells escaping the TGF-β1 action of inhibited proliferation.
To study the role of TGF-β1 and its type Ⅱ receptor in pathogenesis of hepatocellular carcinoma. Methods: The TGF-β1 and its type Ⅱ receptor in the tumor and surrounding tissue in 30 cases with hepatocellular carcinoma were investigated. Results: Levels of protein and mRNA were detected by immuno-histochemistry, hybridization and semi-quantitative analysis. The results showed that TGF-β1 in the tumor and surrounding tissue was overexpressed, and the levels of latter was even higher. TGF-β1 type Ⅱ receptor expressed in surrounding tissue, but it could not be found in most tumor tissues. Conclusion: The lack of TGF-β receptor in the tumor cell may be the way of cancer cells escaping the TGF-β1 action of inhibited proliferation.
2000, 7(1):53-55.
DOI: 10.3872/j.issn.1007-385X.2000.1.016
Abstract:
To construct TCR V β nucleic acid vaccine. Methods: A fragment of TCR V β gene was amplified by RT-PCR from a human T lymphoma cell line, Jurkat, and cloned into eukaryotic expression plasmid pcDNA3 by gene recombination. After sequencing, the recombinant plasmid was transfected into SP2/0 cells and its expression was detected on mRNA level. BALB/c mice were immunized with pcDNA3 and the recombinant plasmid by intramuscular routes. Antiserum was collected and detected by immunocytochemistry method. Results: A fragment of TCR V β gene from Jurkat cells was obtained and proved to be identical to published sequence. A recombinant plasmid pcDNA3-TCR V β was constructed. Its expression was detectable on mRNA level after being transfected into SP2/0 cells. Antiserum from mice immunized with pcDNA3-TCR V β reacted strongly with Jurkat cells and SP2/0 cells transfected by pcDNA3-TCR V β, while shows no reaction on CEM cells expressing TCRγδ and SP2/0 cells. Antisera from normal mice and mice immunized with pcDNA3 were both negative on Jurkat cells. The results of immunocytochemistry indicated that BALB/c mice immunized with pcDNA3-TCR V β produced specific antibody to Jurkat TCR V β. Conclusion: The TCR V β nucleic acid vaccine we constructed can induce humoral immunity.
To construct TCR V β nucleic acid vaccine. Methods: A fragment of TCR V β gene was amplified by RT-PCR from a human T lymphoma cell line, Jurkat, and cloned into eukaryotic expression plasmid pcDNA3 by gene recombination. After sequencing, the recombinant plasmid was transfected into SP2/0 cells and its expression was detected on mRNA level. BALB/c mice were immunized with pcDNA3 and the recombinant plasmid by intramuscular routes. Antiserum was collected and detected by immunocytochemistry method. Results: A fragment of TCR V β gene from Jurkat cells was obtained and proved to be identical to published sequence. A recombinant plasmid pcDNA3-TCR V β was constructed. Its expression was detectable on mRNA level after being transfected into SP2/0 cells. Antiserum from mice immunized with pcDNA3-TCR V β reacted strongly with Jurkat cells and SP2/0 cells transfected by pcDNA3-TCR V β, while shows no reaction on CEM cells expressing TCRγδ and SP2/0 cells. Antisera from normal mice and mice immunized with pcDNA3 were both negative on Jurkat cells. The results of immunocytochemistry indicated that BALB/c mice immunized with pcDNA3-TCR V β produced specific antibody to Jurkat TCR V β. Conclusion: The TCR V β nucleic acid vaccine we constructed can induce humoral immunity.
2000, 7(1):56-59.
DOI: 10.3872/j.issn.1007-385X.2000.1.017
Abstract:
To investigate the effects of PML-RARα antisense on the growth，cell cycle and apoptosis of NB4 cells. Methods： Cell apoptosis and cell cycle were detected by flow cytometry, and leukemic colony forming unit was exammed by methylcellulose assays. Results： PML-RARα antisence （FUAS） could inhibited the growth, and formation of acute myelocytic leukemia colony forming unit （AML-CFU）in NB4 cells； GM-CSF could enhance AML-CFU formation and antagonized the effect of FUAS on AML-CFU formation inhibition. The shorter the time of incubation with FUAS was, the more markedly GM-CSF enhanced the AML-CFU formation. At 7 days, 9 days of treatment with FUAS percentage of apoptotic cells was 41.5%, 34.2% respectively. Reduction of S phase and increase of G2/M phase at 5 days, significant increase of G0/G1 phase with S phase at 7 days. Conclusion: PML-RARα antisence can inhibit the cell growth and induce cell apoptosis of NB4 cells.
To investigate the effects of PML-RARα antisense on the growth，cell cycle and apoptosis of NB4 cells. Methods： Cell apoptosis and cell cycle were detected by flow cytometry, and leukemic colony forming unit was exammed by methylcellulose assays. Results： PML-RARα antisence （FUAS） could inhibited the growth, and formation of acute myelocytic leukemia colony forming unit （AML-CFU）in NB4 cells； GM-CSF could enhance AML-CFU formation and antagonized the effect of FUAS on AML-CFU formation inhibition. The shorter the time of incubation with FUAS was, the more markedly GM-CSF enhanced the AML-CFU formation. At 7 days, 9 days of treatment with FUAS percentage of apoptotic cells was 41.5%, 34.2% respectively. Reduction of S phase and increase of G2/M phase at 5 days, significant increase of G0/G1 phase with S phase at 7 days. Conclusion: PML-RARα antisence can inhibit the cell growth and induce cell apoptosis of NB4 cells.
2000, 7(1):60-62.
DOI: 10.3872/j.issn.1007-385X.2000.1.018
Abstract:
To explore the methods of inducing apoptosis in human bladder cancer multidrug resistance cells. Methods： Expression of Fas antigen was detected with immunocytochemistry on BIU-87/ADM cells, and the effect of anti-APO-1 antibody was examined with in situ DNA terminal deoxynucleotydyl transferase assay (TDT)，phase-contrast and electron microscope techniques. Results： We found that Fas antigen was highly expressed on BIU-87/ADM cell surface, and after treatment with anti-APO-1 antibody, survival rate of BIU-87/ADM cells was decreased, apoptotic morphological changes and TDT positive cells were found. Conclusion： These results suggested that anti-APO-1 antibody could induce apoptosis of BIU-87/ADM cells by combining Fas antigen.
To explore the methods of inducing apoptosis in human bladder cancer multidrug resistance cells. Methods： Expression of Fas antigen was detected with immunocytochemistry on BIU-87/ADM cells, and the effect of anti-APO-1 antibody was examined with in situ DNA terminal deoxynucleotydyl transferase assay (TDT)，phase-contrast and electron microscope techniques. Results： We found that Fas antigen was highly expressed on BIU-87/ADM cell surface, and after treatment with anti-APO-1 antibody, survival rate of BIU-87/ADM cells was decreased, apoptotic morphological changes and TDT positive cells were found. Conclusion： These results suggested that anti-APO-1 antibody could induce apoptosis of BIU-87/ADM cells by combining Fas antigen.
2000, 7(1):63-64.
DOI: 10.3872/j.issn.1007-385X.2000.1.019
Abstract:
Objective: To study the efficiency of radio-immuno-therapy (RIT) with 131I-C50 to precaution and treatment of tumor metastasis and develop a new metastasis suppression method.Methods: The effect was judged through body weight loss, tumor burden, life-span, and metastat- ic. Compared among the multiple low-dose RIT, chemotherapy, surgical resection and controls in different administration time after implantation of tumor and resection of tumor. Results: When the treatment was initiated at 7th day after implantation, the tumor suppression effects of treatment groups were significantly better than those of controls, with the best one in multiple low-dose RIT. Better results were achieved by starting treat- ment earlier. Surgical resection at 15th day after implantation, multiple low-dose RIT groups gained longer life-span and less metastatic nodules than others. Early operation improved therapeutic efficacy, especially in multiple low-dose RIT. Conclusion: Multiple low-dose RIT alone is bet- ter than chemotherapy and surgical treating. Early doing RIT is better than ther. One low-dose RIT Plus with operation had even better tumor suppression effect that RIT alone. Low-dose RIT co-operated with operation as early as possible can enhance the effects of treatment.
Objective: To study the efficiency of radio-immuno-therapy (RIT) with 131I-C50 to precaution and treatment of tumor metastasis and develop a new metastasis suppression method.Methods: The effect was judged through body weight loss, tumor burden, life-span, and metastat- ic. Compared among the multiple low-dose RIT, chemotherapy, surgical resection and controls in different administration time after implantation of tumor and resection of tumor. Results: When the treatment was initiated at 7th day after implantation, the tumor suppression effects of treatment groups were significantly better than those of controls, with the best one in multiple low-dose RIT. Better results were achieved by starting treat- ment earlier. Surgical resection at 15th day after implantation, multiple low-dose RIT groups gained longer life-span and less metastatic nodules than others. Early operation improved therapeutic efficacy, especially in multiple low-dose RIT. Conclusion: Multiple low-dose RIT alone is bet- ter than chemotherapy and surgical treating. Early doing RIT is better than ther. One low-dose RIT Plus with operation had even better tumor suppression effect that RIT alone. Low-dose RIT co-operated with operation as early as possible can enhance the effects of treatment.
2000, 7(1):65-66.
DOI: 10.3872/j.issn.1007-385X.2000.1.020
Abstract:
Objective: To investigate the variation in vitro bystander effect among human lung cancer cell lines aller transfection of pCEAcd-tk. Methods: pCEAcd-tk was extracted and transfected to six human lung cancer cell lines. Their sensitivity to 5- fluorocytosine (5-FC) and ganciclovir (GCV) was measured by MTT assay. Their bystander effect was also detendned. Re- sults: 1. The efficiency of lipid mediated gene transfer was very low; 2. Two out of six human lung cancer cell lines(SPCA-1, A549) have a high sensitivity to 5-FC and GCV; 3. Bystander effect: Two out of six human cancer cell lines(SPCA-1,SC) have a good bystander effect, and one out of them(A549) has a intermediate bystander effect, but three out of them (GM-82, HIAMP and L-78)have a poor bystander effect. Conclusion: There are a variation in vitro bystander effect in six human lung cancer cell lines after transfected with pCEAcd-tk.
Objective: To investigate the variation in vitro bystander effect among human lung cancer cell lines aller transfection of pCEAcd-tk. Methods: pCEAcd-tk was extracted and transfected to six human lung cancer cell lines. Their sensitivity to 5- fluorocytosine (5-FC) and ganciclovir (GCV) was measured by MTT assay. Their bystander effect was also detendned. Re- sults: 1. The efficiency of lipid mediated gene transfer was very low; 2. Two out of six human lung cancer cell lines(SPCA-1, A549) have a high sensitivity to 5-FC and GCV; 3. Bystander effect: Two out of six human cancer cell lines(SPCA-1,SC) have a good bystander effect, and one out of them(A549) has a intermediate bystander effect, but three out of them (GM-82, HIAMP and L-78)have a poor bystander effect. Conclusion: There are a variation in vitro bystander effect in six human lung cancer cell lines after transfected with pCEAcd-tk.
2000, 7(1):69-70.
DOI: 10.3872/j.issn.1007-385X.2000.1.022
Abstract:
Objective: To study the vaccine potency of gene-modified tumor cells, we have constructed IL-12, H7-1 and GM-CSF express vector using retrovirus. Methods: It was transfected into EL-4 thymic lylmphoma cells respectively and the effect of gene transduction on anti-tumor immunity were investigated. Results: The tumorigenicity of EL-4/IL-12 transfectant in C57BI/6 syn- ergistical mice was decreased significantly after implanted with EL-4/IL-12 transfectant compaired with EL-4/Wt or EL-4/Neo groups (P<0. 01). The systemic protective immunity was induced against the challenge with EL-4/Wt (in 10/15 mice) after the rejection of EL-4/IL-12 in the experiment, a stronger CTL activity against EL-4/Wt cells and a weak killer activity against syn- geneic Lewis Lung Carcinoma cells were obtained in 51Cr release assay. In vivo depletion analysis suggested that the decreased tumorigenicity mainly depended on CD4 , CD8 and NK cells. Therapeutic vaccines with EL-4/IL-12 cells could retard the growth to estabished EL-4/Wt tumors significantly compared with those of EL-4/Neo(P < 0 .005 ), combination of therapeutic vaccines of EL-4/IL-12 and EL-4/B7-1 result in the enhanced the therapeutic effect of each single transficants (P < 0 .005) in this experimental model. Conclution: These studies suggested that immunogene treatment using IL-12 is effective in hematopoi- etic malignancy, and combination of IL-12 with B7-1 have a aplication value in human cancer treatment in the near future.
Objective: To study the vaccine potency of gene-modified tumor cells, we have constructed IL-12, H7-1 and GM-CSF express vector using retrovirus. Methods: It was transfected into EL-4 thymic lylmphoma cells respectively and the effect of gene transduction on anti-tumor immunity were investigated. Results: The tumorigenicity of EL-4/IL-12 transfectant in C57BI/6 syn- ergistical mice was decreased significantly after implanted with EL-4/IL-12 transfectant compaired with EL-4/Wt or EL-4/Neo groups (P<0. 01). The systemic protective immunity was induced against the challenge with EL-4/Wt (in 10/15 mice) after the rejection of EL-4/IL-12 in the experiment, a stronger CTL activity against EL-4/Wt cells and a weak killer activity against syn- geneic Lewis Lung Carcinoma cells were obtained in 51Cr release assay. In vivo depletion analysis suggested that the decreased tumorigenicity mainly depended on CD4 , CD8 and NK cells. Therapeutic vaccines with EL-4/IL-12 cells could retard the growth to estabished EL-4/Wt tumors significantly compared with those of EL-4/Neo(P < 0 .005 ), combination of therapeutic vaccines of EL-4/IL-12 and EL-4/B7-1 result in the enhanced the therapeutic effect of each single transficants (P < 0 .005) in this experimental model. Conclution: These studies suggested that immunogene treatment using IL-12 is effective in hematopoi- etic malignancy, and combination of IL-12 with B7-1 have a aplication value in human cancer treatment in the near future.
2000, 7(1):73-75.
DOI: 10.3872/j.issn.1007-385X.2000.1.024
Abstract:
抗体导向酶——前药疗法(antibdy-directed enzyme prodrug therapy, ADEPT)是利用抗体作为载体将前药的专一性活化酶选择性地结合于肿瘤部位，使无毒性的前药可以区域特异性地在肿瘤组织内转化为活性细胞毒分子，杀伤肿瘤。利用不同的活化酶/前药对已建立了多种ADEPT系统，羧肽酶G 2/MMC1系统已用于临床Ⅰ期试验。本文就ADEPT研究和应用的最新进展作一综述。
抗体导向酶——前药疗法(antibdy-directed enzyme prodrug therapy, ADEPT)是利用抗体作为载体将前药的专一性活化酶选择性地结合于肿瘤部位，使无毒性的前药可以区域特异性地在肿瘤组织内转化为活性细胞毒分子，杀伤肿瘤。利用不同的活化酶/前药对已建立了多种ADEPT系统，羧肽酶G 2/MMC1系统已用于临床Ⅰ期试验。本文就ADEPT研究和应用的最新进展作一综述。
联系方式
- 《中国肿瘤生物治疗杂志》
- 1994年创刊
- 主办单位：Chinese Society of Immunology, Chinese Anti-cancer Association
- 邮编：200433
- 电话：021-81871002-22
- 电子邮箱：cjcb@biother.cn
- 网址:http://www.biother.cn
- 刊号：ISSN 1007-385X
- CN 31-1725/R
- 国内定价： ￥20元/册
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Chinese Journal Of Cancer Biotheray ® 2025 All Rights Reserved
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Chinese Journal Of Cancer Biotheray ® 2025 All Rights Reserved
Supported by:Beijing E-Tiller Technology Development Co., Ltd.