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Volume 12,Issue 1,2005 Table of Contents
2005, 12(1):9-12.
DOI: 10.3872/j.issn.1007-385X.2005.1.003
Abstract:
Objective:To develop a peptide vaccine based on MG7-Ag mimotope of gastric cancer using new nano-technology and valuate its efficacy and protective effect. Methods:Encapsulate synthesized MG7 mimotope peptide into nanoemulsion using magnetic and ultrasonic technique. Dialyse method was used to sterilize, HPLC to determine encapsulation efficiency. Select CpG motif as immune adjuvant, Balb/c mice were immunized intravenously with MG7 nanovaccine , with empty nanoemulsion and phosphate buffered saline (PBS) as control. Serum titer of MG7 antibody was determined by ELISA assay. ELISPOT was performed to test the cytotoxicity of spleen cells. The protective effect of the nanovaccine was evaluated by tumor cell challenge assay.Results: G7 nanovaccine was successfully constructed with high stability and encapsulation efficiency. It could induce both cellular and humour immune response to MG7-Ag in mice. Tumor cell challenge shown that the tumor masses formed in the mice immunized with nanovaccine markedly smaller than those formed in the mice of control groups. 2 out of 8 immunized mice were tumor free, while none in the control groups was protected. Conclusion: Nanovaccine based on the MG7-Ag mimotope is immunogenic which can induce specific immunity response against tumor in mice. And the vaccine is partially protective.
Objective:To develop a peptide vaccine based on MG7-Ag mimotope of gastric cancer using new nano-technology and valuate its efficacy and protective effect. Methods:Encapsulate synthesized MG7 mimotope peptide into nanoemulsion using magnetic and ultrasonic technique. Dialyse method was used to sterilize, HPLC to determine encapsulation efficiency. Select CpG motif as immune adjuvant, Balb/c mice were immunized intravenously with MG7 nanovaccine , with empty nanoemulsion and phosphate buffered saline (PBS) as control. Serum titer of MG7 antibody was determined by ELISA assay. ELISPOT was performed to test the cytotoxicity of spleen cells. The protective effect of the nanovaccine was evaluated by tumor cell challenge assay.Results: G7 nanovaccine was successfully constructed with high stability and encapsulation efficiency. It could induce both cellular and humour immune response to MG7-Ag in mice. Tumor cell challenge shown that the tumor masses formed in the mice immunized with nanovaccine markedly smaller than those formed in the mice of control groups. 2 out of 8 immunized mice were tumor free, while none in the control groups was protected. Conclusion: Nanovaccine based on the MG7-Ag mimotope is immunogenic which can induce specific immunity response against tumor in mice. And the vaccine is partially protective.
2005, 12(1):13-18.
DOI: 10.3872/j.issn.1007-385X.2005.1.004
Abstract:
Objective: To obtain phage-displayed ScFv library directly against prostate carcinoma cells, and select antibodies binding to prostate carcinoma cells specifically, so as to lay a foundation for developing diagnostic agents and clinical therapies of prostate carcinoma.Methods:Balb/c mice were immunized i.p .with purified membrane protein mixture of prostate carcinoma cells PC3, DU145. mRNA was isolated from the spleens of immunized mice, heavy and light chain genes （VH and VL） of antibody were amplified separately by RT-PCR and assembled into ScFv gene with a specially constructed linker DNA., the ScFv gene was ligated into the phagemid vector pCANTAB 5E and the ligated sample was transformed into competent E. coli TG1. The transformed cells were infected with M13KO7 helper phage to yield recombinant phage. After five rounds of panning with PC3 cells, the positive clones were selected with the ELISA from the enriched phages. Results: A ScFv library of 3.5×10 6 was obtained and one phage-ScFv which can bind specifically PC3 cells was found. Conclusions: A prostate carcinoma specific antibody was identified ,which paves a way for study of prostate carcinoma.
Objective: To obtain phage-displayed ScFv library directly against prostate carcinoma cells, and select antibodies binding to prostate carcinoma cells specifically, so as to lay a foundation for developing diagnostic agents and clinical therapies of prostate carcinoma.Methods:Balb/c mice were immunized i.p .with purified membrane protein mixture of prostate carcinoma cells PC3, DU145. mRNA was isolated from the spleens of immunized mice, heavy and light chain genes （VH and VL） of antibody were amplified separately by RT-PCR and assembled into ScFv gene with a specially constructed linker DNA., the ScFv gene was ligated into the phagemid vector pCANTAB 5E and the ligated sample was transformed into competent E. coli TG1. The transformed cells were infected with M13KO7 helper phage to yield recombinant phage. After five rounds of panning with PC3 cells, the positive clones were selected with the ELISA from the enriched phages. Results: A ScFv library of 3.5×10 6 was obtained and one phage-ScFv which can bind specifically PC3 cells was found. Conclusions: A prostate carcinoma specific antibody was identified ,which paves a way for study of prostate carcinoma.
2005, 12(1):19-22.
DOI: 10.3872/j.issn.1007-385X.2005.1.005
Abstract:
Objective: To evaluate the therapeutic effect of combination of the human papillomavirus 16 (HPV16) E6/E7 fusion protein vaccine with radiotherapy on the HPV16-positive cervical cancer. Methods: Twenty-four female C57BL/6 mice 7 days after inoculation of tumor cells on the right hind legs were randomly divided into four groups: The control group (C, n=6); Immunotherapy group (IM, n=6); Radiotherapy group (RA, n=6); Immunotherapy+Radiotherapy (IM+RA, n=6). The growth rates of tumors and survival time were recorded and compared with each other. The apoptotic tumor cells in situ were detected by TUNEL assay. Results:The growth rates of tumors in IM+RA- group were lower than other groups. The mean survival time in IM+RA-group was longer than those in IM-, RA- and C-groups, but the difference between IM- and C- group was not significant. The result of TUNEL assay indicated that there were more apoptotic tumor cells in IM+RA- group than that in other groups. Conclusion:The combination of HPV16E6/E7 fusion protein vaccine with low dose radiotherapy exerted synergic antitumor efficacy, which might provide a new approach for the treatment of cervical cancer.
Objective: To evaluate the therapeutic effect of combination of the human papillomavirus 16 (HPV16) E6/E7 fusion protein vaccine with radiotherapy on the HPV16-positive cervical cancer. Methods: Twenty-four female C57BL/6 mice 7 days after inoculation of tumor cells on the right hind legs were randomly divided into four groups: The control group (C, n=6); Immunotherapy group (IM, n=6); Radiotherapy group (RA, n=6); Immunotherapy+Radiotherapy (IM+RA, n=6). The growth rates of tumors and survival time were recorded and compared with each other. The apoptotic tumor cells in situ were detected by TUNEL assay. Results:The growth rates of tumors in IM+RA- group were lower than other groups. The mean survival time in IM+RA-group was longer than those in IM-, RA- and C-groups, but the difference between IM- and C- group was not significant. The result of TUNEL assay indicated that there were more apoptotic tumor cells in IM+RA- group than that in other groups. Conclusion:The combination of HPV16E6/E7 fusion protein vaccine with low dose radiotherapy exerted synergic antitumor efficacy, which might provide a new approach for the treatment of cervical cancer.
2005, 12(1):23-27.
DOI: 10.3872/j.issn.1007-385X.2005.1.006
Abstract:
Objective：To evaluate the sensibility of apoptosis to TRAIL on the residual H22 tumor cells after chemotherapeutic agents treatment and the synergic therapeutic efficiency of pCH510, pTRAIL, and pES in combination with chemotherapeutic agent on mice tumor. Methods：Carrying full length of TRAIL gene, the eukaryotic expression plasmid of pTRAIL was transferred into BHK cells. The mouse hepatocellular carcinoma cell line of H22 which had been treated with ADM, MMC, or 5-FU were mixed with BHK cells. The inhibitory effect of pTRAIL in combination with chemotherapeutic agents was detected by MTT method. The percentage of apoptotic cells and cell cycle of residual H22 cells were analyzed by flow cytometry. The tumor model was made by inoculated with H22 hepatocarcinoma cells in mice. After injection of pCH510, pTRAIL, pES or MMC into intratumor, The therapeutic effects on tumor growth were assessed. Results: pTRAIL could inhibit the growth of H22 tumor cells and induce them to apoptosis. The percentage of apoptotic cells of TRAIL in combination with ADM, MMC, or 5-FU on H22 tumor cells was 28.1%（P<0.01）, 22.5% (P<0.01), and 47% (P>0.05). The tumor was effectively inhibited by MMC+pCH510+pTRAIL+pES . And the percentage of tumorogenesis was 37.5%. Residual tumor cells were scattered and mixed with immune cells from histological detection. Conclusion: Residual H22 tumor cells after ADM or MMC treatment is apt to be induced to apoptosis by pTRAIL. MMC +pCH510+pTRAIL+pES is a great powerful modality for tumor synergic treatment.
Objective：To evaluate the sensibility of apoptosis to TRAIL on the residual H22 tumor cells after chemotherapeutic agents treatment and the synergic therapeutic efficiency of pCH510, pTRAIL, and pES in combination with chemotherapeutic agent on mice tumor. Methods：Carrying full length of TRAIL gene, the eukaryotic expression plasmid of pTRAIL was transferred into BHK cells. The mouse hepatocellular carcinoma cell line of H22 which had been treated with ADM, MMC, or 5-FU were mixed with BHK cells. The inhibitory effect of pTRAIL in combination with chemotherapeutic agents was detected by MTT method. The percentage of apoptotic cells and cell cycle of residual H22 cells were analyzed by flow cytometry. The tumor model was made by inoculated with H22 hepatocarcinoma cells in mice. After injection of pCH510, pTRAIL, pES or MMC into intratumor, The therapeutic effects on tumor growth were assessed. Results: pTRAIL could inhibit the growth of H22 tumor cells and induce them to apoptosis. The percentage of apoptotic cells of TRAIL in combination with ADM, MMC, or 5-FU on H22 tumor cells was 28.1%（P<0.01）, 22.5% (P<0.01), and 47% (P>0.05). The tumor was effectively inhibited by MMC+pCH510+pTRAIL+pES . And the percentage of tumorogenesis was 37.5%. Residual tumor cells were scattered and mixed with immune cells from histological detection. Conclusion: Residual H22 tumor cells after ADM or MMC treatment is apt to be induced to apoptosis by pTRAIL. MMC +pCH510+pTRAIL+pES is a great powerful modality for tumor synergic treatment.
LOU Guo-liang
,
LIU Feng
,
LI Yong-mei
,
CHEN Li
,
HUANG Zheng-xia
,
YANG Hong-bin
,
WANG Liang-hua
,
JIAO Bing-hua
2005, 12(1):28-32.
DOI: 10.3872/j.issn.1007-385X.2005.1.007
Abstract:
Objective：To alleviate graft versus host disease(GVHD) via depletion of alloreactive cells from haematopoietic stem cell grafts. Methods：Balb/c mice dendritic cells genetically engineered to express FasL were cultured with C57BL/6 mice stem cell grafts, and the modified stem cell grafts were used in a C57BL/6 to Balb/c mice GVHD model system (H-2b→H-2d). Then the GVHD clinical manifestations(diarrhea, depilate, lymphocytes infiltration in target tissues) were observed and compared.Results：Recipients that received donor haematopoietic stem cell grafts pretreated with FasL-DC did not develop lethal GVHD, and their survival was also surprisingly prolonged. In contrast, recipients receiving untreated allogeneic grafts displayed all clinical signs of acute GVHD, and died within 30 days after transplantation.Conclusion：DC transfected with FasL gene can prevent GVHD by selective removal of alloreactive cells from haematopoietic stem cell grafts.
Objective：To alleviate graft versus host disease(GVHD) via depletion of alloreactive cells from haematopoietic stem cell grafts. Methods：Balb/c mice dendritic cells genetically engineered to express FasL were cultured with C57BL/6 mice stem cell grafts, and the modified stem cell grafts were used in a C57BL/6 to Balb/c mice GVHD model system (H-2b→H-2d). Then the GVHD clinical manifestations(diarrhea, depilate, lymphocytes infiltration in target tissues) were observed and compared.Results：Recipients that received donor haematopoietic stem cell grafts pretreated with FasL-DC did not develop lethal GVHD, and their survival was also surprisingly prolonged. In contrast, recipients receiving untreated allogeneic grafts displayed all clinical signs of acute GVHD, and died within 30 days after transplantation.Conclusion：DC transfected with FasL gene can prevent GVHD by selective removal of alloreactive cells from haematopoietic stem cell grafts.
2005, 12(1):33-36.
DOI: 10.3872/j.issn.1007-385X.2005.1.008
Abstract:
Objective:To investigate whether the plasmid γ1neo-hgp100 could be expressed and presented in vitro and could protect the immunized mice from B16F10 challenge in vivo.Methods: γ1neo-hgp100 plasmid was constructed in which the DNA sequence encoding hgp100 CTL epitope inserted into CDR3 of γ1-neo vector. The expression of antibodized antigen and IFN-γ in supernatant was measured by ELISA respectively after transfection J558L with γ1neo-hgp100 and further co-culture of J588L transfacted with γ1 neo-hgp100 and pmel TCR transgenic T cell. After introspleenic inoculation of γ1neo-hgp100, the protective efficacy of the gene vaccine was observed by means of measuring the tumor area every two days. Results: γ1neo-hgp100 could be expressed and presented in vitro, the immunogenecity of CTL epitope of hgp100 was strong enough and could activate gp100 specific T cell, the mice immunized with the gene vaccine could resist the tumor challenging in vivo. The mean survival time was prolonged to 36 days, compared to control group(P<005). Conclusion: γ1neo-hgp100 could be expressed and presented in vitro and protect mice from tumor challenging.
Objective:To investigate whether the plasmid γ1neo-hgp100 could be expressed and presented in vitro and could protect the immunized mice from B16F10 challenge in vivo.Methods: γ1neo-hgp100 plasmid was constructed in which the DNA sequence encoding hgp100 CTL epitope inserted into CDR3 of γ1-neo vector. The expression of antibodized antigen and IFN-γ in supernatant was measured by ELISA respectively after transfection J558L with γ1neo-hgp100 and further co-culture of J588L transfacted with γ1 neo-hgp100 and pmel TCR transgenic T cell. After introspleenic inoculation of γ1neo-hgp100, the protective efficacy of the gene vaccine was observed by means of measuring the tumor area every two days. Results: γ1neo-hgp100 could be expressed and presented in vitro, the immunogenecity of CTL epitope of hgp100 was strong enough and could activate gp100 specific T cell, the mice immunized with the gene vaccine could resist the tumor challenging in vivo. The mean survival time was prolonged to 36 days, compared to control group(P<005). Conclusion: γ1neo-hgp100 could be expressed and presented in vitro and protect mice from tumor challenging.
2005, 12(1):37-40.
DOI: 10.3872/j.issn.1007-385X.2005.1.009
Abstract:
Objective: To transfer the gene of n-3 fatty acid desaturase fat-1 into human breast cancer cell QMR2 by adenovirus vector and study the effect of the gene on proliferation of QMR2 cells. Methods: The gene fat-1 was cloned into the shuttle vector of adenovirus, and homologously recombined with an adenoviral backbone vector (pAdEasy 1) to generate the recombinant adenovirus Ad.GFP.fat1; the virus was packaged in 293 cells, inoculated on the breast cancer cells QMR2; total RNA of the cells was hybridized with antisense RNA of fat-1 mRNA by Northern to analyze the expression of fat-1; the effect of fat-1 on the proliferation of QMR2 cells was analyzed by Flow Cytometry; the content of n-6 PUFAs/n-3 PUFAs was analyzed by Gas Chromatography.Results:The high titer recombinant virus was got through DNA recombinant; the fat-1 mRNA appeared in breast cancer cell QMR2 after virus Ad.GFP. fat1 infected the cells for 2 days; compared with the control cells (Ad.GFP), proliferation of QMR2 cells was inhibited by the gene fat-1, decreased by 31%(P<0.05); moreover, fat-1 gene decreased content of n-6 PUFAs/n-3 PUFAs. Conclusion: The gene fat-1 was heterologously expressed in human breast cancer cell QMR2 via adenovirus, and the expression produced an inhibitory effect on proliferation of the cells.
Objective: To transfer the gene of n-3 fatty acid desaturase fat-1 into human breast cancer cell QMR2 by adenovirus vector and study the effect of the gene on proliferation of QMR2 cells. Methods: The gene fat-1 was cloned into the shuttle vector of adenovirus, and homologously recombined with an adenoviral backbone vector (pAdEasy 1) to generate the recombinant adenovirus Ad.GFP.fat1; the virus was packaged in 293 cells, inoculated on the breast cancer cells QMR2; total RNA of the cells was hybridized with antisense RNA of fat-1 mRNA by Northern to analyze the expression of fat-1; the effect of fat-1 on the proliferation of QMR2 cells was analyzed by Flow Cytometry; the content of n-6 PUFAs/n-3 PUFAs was analyzed by Gas Chromatography.Results:The high titer recombinant virus was got through DNA recombinant; the fat-1 mRNA appeared in breast cancer cell QMR2 after virus Ad.GFP. fat1 infected the cells for 2 days; compared with the control cells (Ad.GFP), proliferation of QMR2 cells was inhibited by the gene fat-1, decreased by 31%(P<0.05); moreover, fat-1 gene decreased content of n-6 PUFAs/n-3 PUFAs. Conclusion: The gene fat-1 was heterologously expressed in human breast cancer cell QMR2 via adenovirus, and the expression produced an inhibitory effect on proliferation of the cells.
2005, 12(1):41-45.
DOI: 10.3872/j.issn.1007-385X.2005.1.010
Abstract:
Objective: To investigate the inhibitory effect of VEGF antisense RNA on the growth of human esophageal cancer cells in vitro, in order to further investigate the feasibility of VEGF antisense RNA gene therapy of esophageal cancer. Methods:The plasmid carrying with VEGF antisense cDNA was transfected into esophageal cancer cells, and confirmed its expression by RT-PCR. The expression level of VEGFmRNA and VEGF protein was examined in antisense group by insitu hybridization and immunohistochemistry staining. The cell growth rate was detected by MTT assay. Apoptotic rate in transfected cells was detected by FCM assay. Results: The expression of exogenous antisense VEGFmRNA was confirmed in transfected cells, and the VEGF protein and endogenous VEGFmRNA were dramatically decreased. The growth rate of transfected cells was not inhibited. Apoptotic cells were not found in transfected cells. Latent period of the tumor formation of the antisense group was lengthened, while body weight, volume of tumors was significantly smaller than that of empty vector group and control group. Conclusions: VEGF antisense RNA could decrease the expression of VEGF of esophageal cancer and cell proliferation in vivo, which may apply a useful theory basis for gene therapy of esophageal cancer.
Objective: To investigate the inhibitory effect of VEGF antisense RNA on the growth of human esophageal cancer cells in vitro, in order to further investigate the feasibility of VEGF antisense RNA gene therapy of esophageal cancer. Methods:The plasmid carrying with VEGF antisense cDNA was transfected into esophageal cancer cells, and confirmed its expression by RT-PCR. The expression level of VEGFmRNA and VEGF protein was examined in antisense group by insitu hybridization and immunohistochemistry staining. The cell growth rate was detected by MTT assay. Apoptotic rate in transfected cells was detected by FCM assay. Results: The expression of exogenous antisense VEGFmRNA was confirmed in transfected cells, and the VEGF protein and endogenous VEGFmRNA were dramatically decreased. The growth rate of transfected cells was not inhibited. Apoptotic cells were not found in transfected cells. Latent period of the tumor formation of the antisense group was lengthened, while body weight, volume of tumors was significantly smaller than that of empty vector group and control group. Conclusions: VEGF antisense RNA could decrease the expression of VEGF of esophageal cancer and cell proliferation in vivo, which may apply a useful theory basis for gene therapy of esophageal cancer.
9 The Effect of the Expression Level of CXCR4 on the Metastatic Potential of Human Lung Cancer Cells
2005, 12(1):46-51.
DOI: 10.3872/j.issn.1007-385X.2005.1.011
Abstract:
Objective: To study the effect of the expression levels of Chemokine receptor CXCR4 on the metastatic potential of human lung cancer. Methods:CXCR4 expression was determined by Real-time PCR and FACS. The plasmid DNA containing CXCR4 coding gene or CXCR4 antisense nucleotides fragment was constructed and transfected into 95C or 95D cells with LipofectamineTM2000 reagent respectively, and then the stable transfectants were screened by G418. Migratory responses to SDF-1α were detected by chemotaxis and chemoinvasion assay, MMP-2 activity was determined with zymography, and the ability of adhesion to ECV-304 cells was analyzed by FACS. Results: The surface expression of CXCR4 on lung cancer cells was significantly up or down regulated. Following up-regulation of CXCR4 expression on 95C cells, the migratory response to SDF-1α, MMP-2 activity, and the ability of adhesion to ECV-304 cells were consequently enhanced. Conversely, when CXCR4 expression was down regulated on 95D cells, the migratory response to SDF-1α, MMP-2 activity, and the ability of adhesion to ECV-304 cells were significantly impaired. Conclusion: Up or down regulation of CXCR4 expression enhanced or inhibited the metastatic potential of human lung cancer cells, which implied that CXCR4 expression was closely associated with the metastatic potential of human lung cancer cells.
Objective: To study the effect of the expression levels of Chemokine receptor CXCR4 on the metastatic potential of human lung cancer. Methods:CXCR4 expression was determined by Real-time PCR and FACS. The plasmid DNA containing CXCR4 coding gene or CXCR4 antisense nucleotides fragment was constructed and transfected into 95C or 95D cells with LipofectamineTM2000 reagent respectively, and then the stable transfectants were screened by G418. Migratory responses to SDF-1α were detected by chemotaxis and chemoinvasion assay, MMP-2 activity was determined with zymography, and the ability of adhesion to ECV-304 cells was analyzed by FACS. Results: The surface expression of CXCR4 on lung cancer cells was significantly up or down regulated. Following up-regulation of CXCR4 expression on 95C cells, the migratory response to SDF-1α, MMP-2 activity, and the ability of adhesion to ECV-304 cells were consequently enhanced. Conversely, when CXCR4 expression was down regulated on 95D cells, the migratory response to SDF-1α, MMP-2 activity, and the ability of adhesion to ECV-304 cells were significantly impaired. Conclusion: Up or down regulation of CXCR4 expression enhanced or inhibited the metastatic potential of human lung cancer cells, which implied that CXCR4 expression was closely associated with the metastatic potential of human lung cancer cells.
2005, 12(1):52-56.
DOI: 10.3872/j.issn.1007-385X.2005.1.012
Abstract:
Objective: To develop adriamycin liposome (AL), adriamycin long circulating liposome (ALCL) and adriamycin long circulating temprerature-sensitive liposome (ALTSL) and to study their anti-tumor effects on tumor-bearing mice.Methods:The antitumor activity was observed using the tumor weight as index. The life prolongation rate of mice was calculated according to the tumor-bearing mice survival time. The tissue distribution of adriamycin was determined by HPLC method. Tumor, heart, liver and kidney tissue of the tumor-bearing mice, were sliced and prepared to observe the tissue pathology differences.Results: Compared with free adriamycin, the anti-tumor effects of ALCL and ALTSL were remarkably increased. Their tumor growth inhibitory rates were 57.8% and 67.0% respectively. The study of pharmacokinetics indicated that the adriamycin concentrations were remarkably higher in tumor tissue and blood，lower in heart and lung tissue of ALCL and ALTSL groups when compared with the free ADM group; The pathology slices indicated that tumor cells in the ALTSL group with hyperthermia were mostly destroyed; the cardiac muscle cells in the ALTSL group were similar to the normal cardiac muscle.Conclusion: ALCL and ALTSL remarkably increased the adriamycin concentration on the tumor site, significantly enhanced the anti-tumor effects, decreased the side-effects (such as cytotoxicity) when compared with free ADM, they also significantly prolonged the survival time of the tumor-bearing mice.
Objective: To develop adriamycin liposome (AL), adriamycin long circulating liposome (ALCL) and adriamycin long circulating temprerature-sensitive liposome (ALTSL) and to study their anti-tumor effects on tumor-bearing mice.Methods:The antitumor activity was observed using the tumor weight as index. The life prolongation rate of mice was calculated according to the tumor-bearing mice survival time. The tissue distribution of adriamycin was determined by HPLC method. Tumor, heart, liver and kidney tissue of the tumor-bearing mice, were sliced and prepared to observe the tissue pathology differences.Results: Compared with free adriamycin, the anti-tumor effects of ALCL and ALTSL were remarkably increased. Their tumor growth inhibitory rates were 57.8% and 67.0% respectively. The study of pharmacokinetics indicated that the adriamycin concentrations were remarkably higher in tumor tissue and blood，lower in heart and lung tissue of ALCL and ALTSL groups when compared with the free ADM group; The pathology slices indicated that tumor cells in the ALTSL group with hyperthermia were mostly destroyed; the cardiac muscle cells in the ALTSL group were similar to the normal cardiac muscle.Conclusion: ALCL and ALTSL remarkably increased the adriamycin concentration on the tumor site, significantly enhanced the anti-tumor effects, decreased the side-effects (such as cytotoxicity) when compared with free ADM, they also significantly prolonged the survival time of the tumor-bearing mice.
2005, 12(1):57-60.
DOI: 10.3872/j.issn.1007-385X.2005.1.013
Abstract:
Objective: To investigate whether Vigconic VI-28 capsule, a formulated traditional Chinese medicine containing radix Ginseng, cornu cervi pantotrichum and semen cuscutae, can assist tumor chemotherapy in a mouse model. Methods: Female C57BL/6 mice were s.c. injected with viable Lewis lung cancer (LLC) cells (106 cells/mouse). The mice were then treated with cyclosposphamide (Cyp, 40 mg/kg bodyweight, once every other day). One group was intragastrically given 2% VI-28 (0.5 ml/mouse, every other day) during the course of the chemotherapy. By day 28, the mice were sacrificed and their thymic indices and tumor indices were calculated and compared. Splenocytes were collected for analysis of their immunological status. Histological study was carried to examine the solid tumors. Results: Fourteen days after the injection of LLC cells, solid tumors developed in most of the animals, reaching 1~1.8 cm diameters by 28 th day Compared with mice of the LCC+Cyp group, thymus glands from the LLC+Cyp+VI-28 group were significantly heavier. Splenocytes of the same group responded better to ConA stimulation in vitro. Histochemical examination of the tumor tissues revealed that tumors of the Cyp+VI-28 group were better differentiated (less aggressive) than that of the Cyp group. Conclusion: Vigconic VI-28 capsule can promote recovery of immune system in mice undergoing chemotherapy and help Cyp to control the growth of tumor cells in vivo.
Objective: To investigate whether Vigconic VI-28 capsule, a formulated traditional Chinese medicine containing radix Ginseng, cornu cervi pantotrichum and semen cuscutae, can assist tumor chemotherapy in a mouse model. Methods: Female C57BL/6 mice were s.c. injected with viable Lewis lung cancer (LLC) cells (106 cells/mouse). The mice were then treated with cyclosposphamide (Cyp, 40 mg/kg bodyweight, once every other day). One group was intragastrically given 2% VI-28 (0.5 ml/mouse, every other day) during the course of the chemotherapy. By day 28, the mice were sacrificed and their thymic indices and tumor indices were calculated and compared. Splenocytes were collected for analysis of their immunological status. Histological study was carried to examine the solid tumors. Results: Fourteen days after the injection of LLC cells, solid tumors developed in most of the animals, reaching 1~1.8 cm diameters by 28 th day Compared with mice of the LCC+Cyp group, thymus glands from the LLC+Cyp+VI-28 group were significantly heavier. Splenocytes of the same group responded better to ConA stimulation in vitro. Histochemical examination of the tumor tissues revealed that tumors of the Cyp+VI-28 group were better differentiated (less aggressive) than that of the Cyp group. Conclusion: Vigconic VI-28 capsule can promote recovery of immune system in mice undergoing chemotherapy and help Cyp to control the growth of tumor cells in vivo.
LIU Cheng-xia
,
ZHANG Shang-zhong
,
LI Tie-jun
,
HUANG Li-hua
,
WANG Bing
,
ZHANG Xiao-wei
,
DI Xiu-lan
2005, 12(1):63-64.
DOI: 10.3872/j.issn.1007-385X.2005.1.015
Abstract:
目的：选择性去除骨髓移植物中异基因反应性淋巴细胞，特异性抑制移植物抗宿主病(GVHD)。方法：用携带有FasL基因的重组腺病毒转染Balb／c小鼠来源的树突状细胞(dendritic cells，DC)，并与c57BL／6小鼠骨髓细胞移植物共培养，把经过这种处理的骨髓细胞移植物移植给Balb／c受体小鼠(C57BL／6→Balb／c小鼠GVHD模型，H-2^b-H-2^d)，然后观察、比较各组GVHD表现。结果：致死剂量照射的受体鼠在接受经FasL-DC处理的供体骨髓细胞移植后，没有出现明显的GVHD表现，生存期显著延长，3个月时生存率80％以上。但对照组2周后均出现了明显的GVHD症状，腹泻、脱毛和靶组织淋巴细胞浸润等，生存期没有超过30d。结论：转染。FasL基因的DC可有效去除骨髓移植物中异基因反应性T淋巴细胞，移植用这种方法处理过的骨髓，能够有效抑制GVHD的发生。
目的：选择性去除骨髓移植物中异基因反应性淋巴细胞，特异性抑制移植物抗宿主病(GVHD)。方法：用携带有FasL基因的重组腺病毒转染Balb／c小鼠来源的树突状细胞(dendritic cells，DC)，并与c57BL／6小鼠骨髓细胞移植物共培养，把经过这种处理的骨髓细胞移植物移植给Balb／c受体小鼠(C57BL／6→Balb／c小鼠GVHD模型，H-2^b-H-2^d)，然后观察、比较各组GVHD表现。结果：致死剂量照射的受体鼠在接受经FasL-DC处理的供体骨髓细胞移植后，没有出现明显的GVHD表现，生存期显著延长，3个月时生存率80％以上。但对照组2周后均出现了明显的GVHD症状，腹泻、脱毛和靶组织淋巴细胞浸润等，生存期没有超过30d。结论：转染。FasL基因的DC可有效去除骨髓移植物中异基因反应性T淋巴细胞，移植用这种方法处理过的骨髓，能够有效抑制GVHD的发生。
2005, 12(1):65-66.
DOI: 10.3872/j.issn.1007-385X.2005.1.016
Abstract:
目的：构建含有黑色素肿瘤相关抗原hgp100的CTL表位编码基因质粒γ1neo-hgp100，观察此质粒的体外表达和表达产物的提呈，以及体内基因免疫对黑色素肿瘤攻击的保护效应。方法：将编码hgp100的CTL表位的DNA序列插入抗体化抗原的表达质粒γ1neo中，构建γ1neo-hgp100，体外转染J558L，ELISA检测免疫球蛋白的表达；转染后的J558L与pmel TCR转基因T细胞共育，48h后ELISA检测培养上清中IFN-γ的含量。以γ1neo-hgp100脾内基因免疫C57BL／6小鼠，以B16F10黑色素瘤细胞攻击免疫后小鼠，定期观察肿瘤的生长情况及大小。结果：所构建的γ1neo-hgp100可以在体外表达，表达产物可以被充分的提呈，被提呈的hgp100 CTL表位能有效的活化pmel TCRT细胞分泌IFN-γ。经γ1neo-hgp100免疫的小鼠能显著抵抗B16F10黑色素瘤细胞的攻击，生存时间得以明显延长。结论：γ1neo-hgp100基因可有效的在体外表达并被提呈至T细胞，产生体内免疫保护效应。
目的：构建含有黑色素肿瘤相关抗原hgp100的CTL表位编码基因质粒γ1neo-hgp100，观察此质粒的体外表达和表达产物的提呈，以及体内基因免疫对黑色素肿瘤攻击的保护效应。方法：将编码hgp100的CTL表位的DNA序列插入抗体化抗原的表达质粒γ1neo中，构建γ1neo-hgp100，体外转染J558L，ELISA检测免疫球蛋白的表达；转染后的J558L与pmel TCR转基因T细胞共育，48h后ELISA检测培养上清中IFN-γ的含量。以γ1neo-hgp100脾内基因免疫C57BL／6小鼠，以B16F10黑色素瘤细胞攻击免疫后小鼠，定期观察肿瘤的生长情况及大小。结果：所构建的γ1neo-hgp100可以在体外表达，表达产物可以被充分的提呈，被提呈的hgp100 CTL表位能有效的活化pmel TCRT细胞分泌IFN-γ。经γ1neo-hgp100免疫的小鼠能显著抵抗B16F10黑色素瘤细胞的攻击，生存时间得以明显延长。结论：γ1neo-hgp100基因可有效的在体外表达并被提呈至T细胞，产生体内免疫保护效应。
2005, 12(1):67-68.
DOI: 10.3872/j.issn.1007-385X.2005.1.017
Abstract:
Objective: To transfer the gene of n-3 fatty acid desaturase fat-1 into human breast cancer cell QMR2 by adenovirus vector and study the effect of the gene on proliferation of QMR2 cells. Methods: The gene fat-1 was cloned into the shuttle vector of adenovirus, and homologously recombined with an adenoviral backbone vector ( pAdEasy 1) to generate the recombinant adenovirus Ad. GFP. fall; the virus was packaged in 293 cells, inoculated on the breast cancer cells QMR2; total RNA of the cells was hybridized with antisense RNA of fat-1 mRNA by Northern to analyze the expression of fat-1; the effect of fat-1 on the proliferation of QMR2 cells was analyzed by Flow Cytometry; the content of n-6 PUFAs/n-3 PUFAs was analyzed by Gas Chromatography. Results: The high titer recombinant virus was got through DNA recombinant ; the fat-1 mRNA appeared in breast cancer cell QMR2 after virus Ad. GFP. fat1 infected the cells for 2 days; compared with the control cells (Ad. GFP) , proliferation of QMR2 cells was inhibited by the gene fat-1, decreased by 31% ( P < 0.05); moreover, fat-1 gene decreased content of n-6 PUFAs/n-3 PUFAs. Conclusion: The gene fat - 1 was heter-ologously expressed in human breast cancer cell QMR2 via adenovirus, and the expression produced an inhibitory effect on proliferation of the cells.
Objective: To transfer the gene of n-3 fatty acid desaturase fat-1 into human breast cancer cell QMR2 by adenovirus vector and study the effect of the gene on proliferation of QMR2 cells. Methods: The gene fat-1 was cloned into the shuttle vector of adenovirus, and homologously recombined with an adenoviral backbone vector ( pAdEasy 1) to generate the recombinant adenovirus Ad. GFP. fall; the virus was packaged in 293 cells, inoculated on the breast cancer cells QMR2; total RNA of the cells was hybridized with antisense RNA of fat-1 mRNA by Northern to analyze the expression of fat-1; the effect of fat-1 on the proliferation of QMR2 cells was analyzed by Flow Cytometry; the content of n-6 PUFAs/n-3 PUFAs was analyzed by Gas Chromatography. Results: The high titer recombinant virus was got through DNA recombinant ; the fat-1 mRNA appeared in breast cancer cell QMR2 after virus Ad. GFP. fat1 infected the cells for 2 days; compared with the control cells (Ad. GFP) , proliferation of QMR2 cells was inhibited by the gene fat-1, decreased by 31% ( P < 0.05); moreover, fat-1 gene decreased content of n-6 PUFAs/n-3 PUFAs. Conclusion: The gene fat - 1 was heter-ologously expressed in human breast cancer cell QMR2 via adenovirus, and the expression produced an inhibitory effect on proliferation of the cells.
2005, 12(1):69-71.
DOI: 10.3872/j.issn.1007-385X.2005.1.018
Abstract:
Objective: To investigate the inhibitory effect of VEGF antisense RNA on the growth of human esophageal cancer cells in vitro, in order to further investigate the feasibility of VEGF antisense RNA gene therapy of esophageal cancer. Methods: The plasmid carrying with VEGF antisense cDNA was transfected into esophageal cancer cells, and confirmed its expression by RT-PCR. The expression level of VEGFmRNA and VEGF protein was examined in antisense group by insitu hybridization and immunohistochemistry staining. The cell growth rate was detected by MTT assay. Apoptotic rate in transfected cells was detected by FCM assay. Results: The expression of exogenous antisense VEGFmRNA was confirmed in transfected cells, and the VEGF protein and endogenous VEGFmRNA were dramatically decreased. The growth rate of transfected cells was not inhibited. Apoptotic cells were not found in transfected cells. Latent period of the tumor formation of the antisense group was lengthened, while body weight, volume of tumors was significantly smaller than that of empty vector group and control group. Conclusions: VEGF antisense RNA could decrease the expression of VEGF of e-sophageal cancer and cell proliferation in vivo, which may apply a useful theory basis for gene therapy of esophageal canc-er.
Objective: To investigate the inhibitory effect of VEGF antisense RNA on the growth of human esophageal cancer cells in vitro, in order to further investigate the feasibility of VEGF antisense RNA gene therapy of esophageal cancer. Methods: The plasmid carrying with VEGF antisense cDNA was transfected into esophageal cancer cells, and confirmed its expression by RT-PCR. The expression level of VEGFmRNA and VEGF protein was examined in antisense group by insitu hybridization and immunohistochemistry staining. The cell growth rate was detected by MTT assay. Apoptotic rate in transfected cells was detected by FCM assay. Results: The expression of exogenous antisense VEGFmRNA was confirmed in transfected cells, and the VEGF protein and endogenous VEGFmRNA were dramatically decreased. The growth rate of transfected cells was not inhibited. Apoptotic cells were not found in transfected cells. Latent period of the tumor formation of the antisense group was lengthened, while body weight, volume of tumors was significantly smaller than that of empty vector group and control group. Conclusions: VEGF antisense RNA could decrease the expression of VEGF of e-sophageal cancer and cell proliferation in vivo, which may apply a useful theory basis for gene therapy of esophageal canc-er.
16 The Effect of the Expression Level of CXCR4 on the Metastatic Potential of Human Lung Cancer Cells
2005, 12(1):71-73.
DOI: 10.3872/j.issn.1007-385X.2005.1.019
Abstract:
Stathmin蛋白由于其特有的微管解聚活性，在细胞的增殖和分化及肿瘤发生中有十分重要的作用。抑制Stathmin蛋白的表达已经成为肿瘤基因治疗的新的靶点，并且已经证实Stathmin蛋白能够影响某些作用于微管的化疗药物的疗效，对于指导临床用药有一定的意义。Stathmin蛋白还能够促进神经系统的发育，因此受到更多的关注
Stathmin蛋白由于其特有的微管解聚活性，在细胞的增殖和分化及肿瘤发生中有十分重要的作用。抑制Stathmin蛋白的表达已经成为肿瘤基因治疗的新的靶点，并且已经证实Stathmin蛋白能够影响某些作用于微管的化疗药物的疗效，对于指导临床用药有一定的意义。Stathmin蛋白还能够促进神经系统的发育，因此受到更多的关注
2005, 12(1):76-79.
DOI: 10.3872/j.issn.1007-385X.2005.1.021
Abstract:
Objective: To develop adriamycin liposome (AL) , adriamycin long circulating liposome (ALCL) and adriamycin long circulating temprerature-sensitive liposome ( ALTSL) and to study their anti-tumor effects on tumor-bearing mice. Methods: The antitumor activity was observed using the tumor weight as index. The life prolongation rate of mice was calculated according to the tumor-bearing mice survival time. The tissue distribution of adriamycin was determined by HPLC method. Tumor, heart, liver and kidney tissue of the tumor-bearing mice, were sliced and prepared to observe the tissue pathology differences. Results: Compared with free adriamycin, the anti-tumor effects of ALCL and ALTSL were remarkably increased. Their tumor growth inhibitory rates were 57. 8% and 67. 0% respectively. The study of pharmacoki-netics indicated that the adriamycin concentrations were remarkably higher in tumor tissue and blood,lower in heart and lung tissue of ALCL and ALTSL groups when compared with the free ADM group; The pathology slices indicated that tumor cells in the ALTSL group with hyperthermia were mostly destroyed; the cardiac muscle cells in the ALTSL group were similar to the normal cardiac muscle. Conclusion: ALCL and ALTSL remarkably increased the adriamycin concentration on the tumor site, significantly enhanced the anti-tumor effects, decreased the side-effects (such as cytotoxicity) when compared with free ADM, they also significantly prolonged the survival time of the tumor-bearing mice.
Objective: To develop adriamycin liposome (AL) , adriamycin long circulating liposome (ALCL) and adriamycin long circulating temprerature-sensitive liposome ( ALTSL) and to study their anti-tumor effects on tumor-bearing mice. Methods: The antitumor activity was observed using the tumor weight as index. The life prolongation rate of mice was calculated according to the tumor-bearing mice survival time. The tissue distribution of adriamycin was determined by HPLC method. Tumor, heart, liver and kidney tissue of the tumor-bearing mice, were sliced and prepared to observe the tissue pathology differences. Results: Compared with free adriamycin, the anti-tumor effects of ALCL and ALTSL were remarkably increased. Their tumor growth inhibitory rates were 57. 8% and 67. 0% respectively. The study of pharmacoki-netics indicated that the adriamycin concentrations were remarkably higher in tumor tissue and blood,lower in heart and lung tissue of ALCL and ALTSL groups when compared with the free ADM group; The pathology slices indicated that tumor cells in the ALTSL group with hyperthermia were mostly destroyed; the cardiac muscle cells in the ALTSL group were similar to the normal cardiac muscle. Conclusion: ALCL and ALTSL remarkably increased the adriamycin concentration on the tumor site, significantly enhanced the anti-tumor effects, decreased the side-effects (such as cytotoxicity) when compared with free ADM, they also significantly prolonged the survival time of the tumor-bearing mice.
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
Supported by:Beijing E-Tiller Technology Development Co., Ltd.