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Volume 11,Issue 1,2004 Table of Contents
YU Chang-zhong
,
LIN Chen
,
ZHANG Hai-zeng
,
ZHA Yuan-yuan
,
LIANG Xiao
,
FU Ming
,
ZHANG Xue-yan
,
WU Min
2004, 11(1):5-9.
DOI: 10.3872/j.issn.1007-385X.2004.1.002
Abstract:
Objective: To assess the preclinical safety of recombinant adenovirus-mediated antisense c-myc infustion of the hepatic vascular of Beagle dogs. Methods: Four 8 to 10 kg healthy Beagle dogs underwent hepatic artery or portal vein cannulation infusion of 1×10 10～1×10 11 plaque forming units recombinant adenovirus vectors. Serial sequenced liver biopsies were taken for microscopic examination and PCR analysis. Venous blood samples were obtained from the dogs for liver, renal function tests and hematology analysis after infusion of days 0, 3, 7, 14, and 21. PCR was used to screen the vital organs for the presence of adenovirus DNA. Microscopic examination of the vital organs was performed to observe the pathogenicity of Ad-ASmyc. ELISA was performed to assay the neutralizing antibody to adenovirus vectors. Results：Ad-ASmyc could infect both kinds of normal human fetal lung cell line 2BS and normal human hepatocyte line L02 effectively, but could not inhibit their growth in vitro. Results of liver, renal function and hematology values were within normal ranges. The adenovirus vectors were present in the liver, spleen, heart, kidney, stomach and skin at days 14 or 21. Microscopic examination revealed no cytopathic effects in distant organs, as was gross pathology, despite the presence of vector. Ad-ASmyc was transferred into hepatocytes persistently and induced dose-dependent inflammation response. Production of anti-adenovirus antibodies appeared at days 7, reached high-level at days 14 and declined at days 21.Conclusions：Ad-ASmyc can be transferred into hepatocellulars via hepatic artery or portal vein and generates no obvious toxicity in Beagle dogs, which suggests the safety of Ad-ASmyc infused into the hepatic vascular for gene therapy.
Objective: To assess the preclinical safety of recombinant adenovirus-mediated antisense c-myc infustion of the hepatic vascular of Beagle dogs. Methods: Four 8 to 10 kg healthy Beagle dogs underwent hepatic artery or portal vein cannulation infusion of 1×10 10～1×10 11 plaque forming units recombinant adenovirus vectors. Serial sequenced liver biopsies were taken for microscopic examination and PCR analysis. Venous blood samples were obtained from the dogs for liver, renal function tests and hematology analysis after infusion of days 0, 3, 7, 14, and 21. PCR was used to screen the vital organs for the presence of adenovirus DNA. Microscopic examination of the vital organs was performed to observe the pathogenicity of Ad-ASmyc. ELISA was performed to assay the neutralizing antibody to adenovirus vectors. Results：Ad-ASmyc could infect both kinds of normal human fetal lung cell line 2BS and normal human hepatocyte line L02 effectively, but could not inhibit their growth in vitro. Results of liver, renal function and hematology values were within normal ranges. The adenovirus vectors were present in the liver, spleen, heart, kidney, stomach and skin at days 14 or 21. Microscopic examination revealed no cytopathic effects in distant organs, as was gross pathology, despite the presence of vector. Ad-ASmyc was transferred into hepatocytes persistently and induced dose-dependent inflammation response. Production of anti-adenovirus antibodies appeared at days 7, reached high-level at days 14 and declined at days 21.Conclusions：Ad-ASmyc can be transferred into hepatocellulars via hepatic artery or portal vein and generates no obvious toxicity in Beagle dogs, which suggests the safety of Ad-ASmyc infused into the hepatic vascular for gene therapy.
2004, 11(1):10-13.
DOI: 10.3872/j.issn.1007-385X.2004.1.003
Abstract:
Objective:To investigate the effect of human Angiopoietin-1 (Ang-1) on the proliferation and apoptosis of human umbilical vein endothelial cell (HUVEC).Methods: The recombinant eukaryotic expression vector pcDNA3.1-V5-HisC-Ang1 was constructed and was used to transiently transfecte to 293 cells. HUVEC was seperated from new born fetel cord. The supernatants from transfected 293 cells were harvested, and then added to HUVEC culture. The proliferation and apoptosis of HUVEC was measured by MTT colorimetry assay analysis, cell counting and flow cytometry, respectively.Results：Proliferation of HUVEC cells cultured with supurnatants of Ang-1 transfected group was higher than those untreated and mock transfected group, results of which were 0.68±0.10, 0.36±0.11, 0.40±0.03 in the MTT assays; and were （15.03±198）×10 4, （10.13±2.06）×10 4 and (8.7±1.73)×10 4 respectively in the cell counts. The apoptotic pereentage of HUVEC cells culturel with supernatants from Ang-1 transfected group was lower than those of untreated or mock transfeeted group, results of which were 6%,21% and 19% respectively in FACS analysis. Conclusions：Human Ang-1 can significacetly pronaete stimulate the proliferation and reduce the apoptosis of endothelial cells.
Objective:To investigate the effect of human Angiopoietin-1 (Ang-1) on the proliferation and apoptosis of human umbilical vein endothelial cell (HUVEC).Methods: The recombinant eukaryotic expression vector pcDNA3.1-V5-HisC-Ang1 was constructed and was used to transiently transfecte to 293 cells. HUVEC was seperated from new born fetel cord. The supernatants from transfected 293 cells were harvested, and then added to HUVEC culture. The proliferation and apoptosis of HUVEC was measured by MTT colorimetry assay analysis, cell counting and flow cytometry, respectively.Results：Proliferation of HUVEC cells cultured with supurnatants of Ang-1 transfected group was higher than those untreated and mock transfected group, results of which were 0.68±0.10, 0.36±0.11, 0.40±0.03 in the MTT assays; and were （15.03±198）×10 4, （10.13±2.06）×10 4 and (8.7±1.73)×10 4 respectively in the cell counts. The apoptotic pereentage of HUVEC cells culturel with supernatants from Ang-1 transfected group was lower than those of untreated or mock transfeeted group, results of which were 6%,21% and 19% respectively in FACS analysis. Conclusions：Human Ang-1 can significacetly pronaete stimulate the proliferation and reduce the apoptosis of endothelial cells.
2004, 11(1):14-17.
DOI: 10.3872/j.issn.1007-385X.2004.1.004
Abstract:
Objective:To evaluate the effects of antisense oligodeoxynucleotides (ODNs) (AS1 complementary to the translation initiation region and AS2 complementary to the coding region) targeted to bcl-2 oncogene on Bcl-2 protein expression and apoptosis of human renal cell carcinoma (RCC) cells.Methods:Expression of bcl-2 mRNA in RCC cell lines was analyzed by reverse transcriptase-polymerase chain reaction (RT-PCR). The ODNs were transfected with Lipofectin into RCC cell lines. The expression of Bcl-2 protein in ACHN tumor cells was examined by Western blot analysis, and the apoptosis of those cells was determined by flow cytometric analysis.Results: Expression of bcl-2 mRNA was detected in all five RCC lines. Transfected bcl-2 antisense ODNs, but not sense ODNs, inhibited Bcl-2 protein expression in ACHN cells. The AS2 antisense ODN showed a superior effect compared with AS1 ODN. The apoptosis of ACHN cell could been induced by bcl-2 antisese ODNs , and percentage of apoptotic cells was noted 32.1% and 43.2% treated with AS1 and AS2, respectively. Conclusions:Treatment of human RCC cells with antisense ODNs targeting bcl-2 gene inhibits expression of Bcl-2 protein and induce apoptosis.
Objective:To evaluate the effects of antisense oligodeoxynucleotides (ODNs) (AS1 complementary to the translation initiation region and AS2 complementary to the coding region) targeted to bcl-2 oncogene on Bcl-2 protein expression and apoptosis of human renal cell carcinoma (RCC) cells.Methods:Expression of bcl-2 mRNA in RCC cell lines was analyzed by reverse transcriptase-polymerase chain reaction (RT-PCR). The ODNs were transfected with Lipofectin into RCC cell lines. The expression of Bcl-2 protein in ACHN tumor cells was examined by Western blot analysis, and the apoptosis of those cells was determined by flow cytometric analysis.Results: Expression of bcl-2 mRNA was detected in all five RCC lines. Transfected bcl-2 antisense ODNs, but not sense ODNs, inhibited Bcl-2 protein expression in ACHN cells. The AS2 antisense ODN showed a superior effect compared with AS1 ODN. The apoptosis of ACHN cell could been induced by bcl-2 antisese ODNs , and percentage of apoptotic cells was noted 32.1% and 43.2% treated with AS1 and AS2, respectively. Conclusions:Treatment of human RCC cells with antisense ODNs targeting bcl-2 gene inhibits expression of Bcl-2 protein and induce apoptosis.
JIANG Ming-hong
,
NIE Ming-ming
,
FANG Guo-en
,
CUI Zhen-fu
,
WU Hong-ping
,
LI Lin-fang
,
WU Meng-chao
,
QIAN Qi-jun
2004, 11(1):18-21.
DOI: 10.3872/j.issn.1007-385X.2004.1.005
Abstract:
Objective：To study the inhibitory effects on gastric cancer by adenoviral transduction of mouse endostatin gene.Methods：Mouse endostatin gene was cloned into the genome of replication-defective adenovirus specific for the tumor cells by virus recombination technology, and then its biological activities were surveyed in vitro. Animal model of gastric cancer bearing nude mice were established to assay the expression of mouse endostatin and inhibition for tumor cell in vivo. Results：To construct the recombinant adenovirus vector pCA13-mEndo and explore its expression in the levels of both mRNA and protein. The protein appearing the anti-angiogenesis activity was tested by chicken chorio-allantoic membrane assay. It can also inhibit tumor to grow and induce tumor cell to apoptosis significantly in vivo of animal. Conclusions:The recombination adenovirus can express biologically active mEndostatin effectively, which results in inhibiting the growth of micro-blood vessels and proliferation slowly. This lays the foundation for the angiogenesis therapy of the solid tumor in clinical application.
Objective：To study the inhibitory effects on gastric cancer by adenoviral transduction of mouse endostatin gene.Methods：Mouse endostatin gene was cloned into the genome of replication-defective adenovirus specific for the tumor cells by virus recombination technology, and then its biological activities were surveyed in vitro. Animal model of gastric cancer bearing nude mice were established to assay the expression of mouse endostatin and inhibition for tumor cell in vivo. Results：To construct the recombinant adenovirus vector pCA13-mEndo and explore its expression in the levels of both mRNA and protein. The protein appearing the anti-angiogenesis activity was tested by chicken chorio-allantoic membrane assay. It can also inhibit tumor to grow and induce tumor cell to apoptosis significantly in vivo of animal. Conclusions:The recombination adenovirus can express biologically active mEndostatin effectively, which results in inhibiting the growth of micro-blood vessels and proliferation slowly. This lays the foundation for the angiogenesis therapy of the solid tumor in clinical application.
2004, 11(1):22-26.
DOI: 10.3872/j.issn.1007-385X.2004.1.006
Abstract:
bjective:To observe the anti-tumor effects of adenovirus carring AFP gene transfected DC using different inculabe administrations.Methods:The immunotherapy effects of AFP-DC vaccine were compared among administration of intra-vein, hypo-dermal and intra-tumor. The protective and therapeutic effects of AFP-DC tumor vaccine were detected by 4 h 51 Cr releasing assay and depleting T cells and NK cells. Results: The therapentic effect of Hypo-dermal was significantly better than assays of the other two ways in inhibiting tumor growth and prolonging the survival period of the mice(P<0.05). AFP-DC tumor vaccine could more effectively induced specific CTL cytotoxicity, protective response, and resistance to the rechallenge of tumor cells. Both CD4+ and CD8+T cells were required for AFP-DC tumor vaccine to induce antitumor immunological rejection. In the effective phase, antitumor immunological rejections were mainly depended on CD8+T cells, and CD4+ T cells were not necessary. Blocking NK cells in the induction and effect phase didn′t have obvious influence on the antitumor immunity, suggesting that NK cells were not required for antitumor immunity induced by AFP-DC tumor vaccine. Conclusion: AFP-DC tumor vaccine by hypo-dermal administration could induce vaccine more efficient antitumor immune responses than intravein or intra-tumor ways in vivo, which provided new ways to the immunotherapy of tumor mediated by DC.
bjective:To observe the anti-tumor effects of adenovirus carring AFP gene transfected DC using different inculabe administrations.Methods:The immunotherapy effects of AFP-DC vaccine were compared among administration of intra-vein, hypo-dermal and intra-tumor. The protective and therapeutic effects of AFP-DC tumor vaccine were detected by 4 h 51 Cr releasing assay and depleting T cells and NK cells. Results: The therapentic effect of Hypo-dermal was significantly better than assays of the other two ways in inhibiting tumor growth and prolonging the survival period of the mice(P<0.05). AFP-DC tumor vaccine could more effectively induced specific CTL cytotoxicity, protective response, and resistance to the rechallenge of tumor cells. Both CD4+ and CD8+T cells were required for AFP-DC tumor vaccine to induce antitumor immunological rejection. In the effective phase, antitumor immunological rejections were mainly depended on CD8+T cells, and CD4+ T cells were not necessary. Blocking NK cells in the induction and effect phase didn′t have obvious influence on the antitumor immunity, suggesting that NK cells were not required for antitumor immunity induced by AFP-DC tumor vaccine. Conclusion: AFP-DC tumor vaccine by hypo-dermal administration could induce vaccine more efficient antitumor immune responses than intravein or intra-tumor ways in vivo, which provided new ways to the immunotherapy of tumor mediated by DC.
2004, 11(1):27-30.
DOI: 10.3872/j.issn.1007-385X.2004.1.007
Abstract:
Objective:To investigate the vaccine potency of GM-CSF anchored B16 tumor cells. Methods:In this study, mGM-CSF was expressed on surface of B16 mice melanoma cells by GPI-modifying. C57BL/6 mice were inoculated with GM-CSF anchored cells and wide-type B16 cells to evaluate whether the GM-CSF anchored cells could elicit a protective and systemtic antitumor response. Results:GM-CSF anchored cells resulted in remarkable loss of tumorgenicity in syngenetic mice. The tumor occurrence rate of GM-CSF anchored B16 cells was 58.8% on C57BL/6 mice with 1×10 6- B16 cells/mice inoculated (n=12) and that of wide-type B16 cells was 100%, The C57BL/6 mice receiving inoculation with 5×10 5 GM-CSF anchored cells/mice never grew tumor. These mice were challenged with wide-type B16 cells, and only a minority of mice grew tumor after wide-type B16 cells inoculation. Conclusion: GM-CSF protein anchored cells could elicit a protective and systemtic antitumor responses.
Objective:To investigate the vaccine potency of GM-CSF anchored B16 tumor cells. Methods:In this study, mGM-CSF was expressed on surface of B16 mice melanoma cells by GPI-modifying. C57BL/6 mice were inoculated with GM-CSF anchored cells and wide-type B16 cells to evaluate whether the GM-CSF anchored cells could elicit a protective and systemtic antitumor response. Results:GM-CSF anchored cells resulted in remarkable loss of tumorgenicity in syngenetic mice. The tumor occurrence rate of GM-CSF anchored B16 cells was 58.8% on C57BL/6 mice with 1×10 6- B16 cells/mice inoculated (n=12) and that of wide-type B16 cells was 100%, The C57BL/6 mice receiving inoculation with 5×10 5 GM-CSF anchored cells/mice never grew tumor. These mice were challenged with wide-type B16 cells, and only a minority of mice grew tumor after wide-type B16 cells inoculation. Conclusion: GM-CSF protein anchored cells could elicit a protective and systemtic antitumor responses.
QU Xun
,
YANG Mei-xiang
,
ZHENG Guang-juan
,
GUO Wen-jing
,
ZHOU Wen
,
LIU De-shan
,
ZHANG Dan
,
ZHANG jing
,
ZHAO Li-xia
,
XIA Li-ying
2004, 11(1):31-35.
DOI: 10.3872/j.issn.1007-385X.2004.1.008
Abstract:
Objective: To study the effect of basil polysaccharide (BP) on tumor cell metastasis and its mechanism through assays of intercellular communication and NK cytotoxicity.Methods: Metastasis of tumor cell ability was assayed by metrigel in vitro and the rate of pulmonary metastasis nodel was caluated. Using SLDT technique to test the effect of BP on recovering the function of cell Gap junction mediated intercellular communication (GJIC). NK cytotoxicity was measured by a standard 4 h 51 Cr-release assay. Results:Invasion and migration of tumor cells decreased significantly after treated with BP in vitro and in vivo.PG cells treated with BP for 8 h could recover the function of GJIC compared with the control. NK activity of mice bearing tumor treated with BP rose significantly. Conclusions: The resutls suggested that BP could inhibit tumor metastasis and might be a new candidate for antitumor metastasis agent.
Objective: To study the effect of basil polysaccharide (BP) on tumor cell metastasis and its mechanism through assays of intercellular communication and NK cytotoxicity.Methods: Metastasis of tumor cell ability was assayed by metrigel in vitro and the rate of pulmonary metastasis nodel was caluated. Using SLDT technique to test the effect of BP on recovering the function of cell Gap junction mediated intercellular communication (GJIC). NK cytotoxicity was measured by a standard 4 h 51 Cr-release assay. Results:Invasion and migration of tumor cells decreased significantly after treated with BP in vitro and in vivo.PG cells treated with BP for 8 h could recover the function of GJIC compared with the control. NK activity of mice bearing tumor treated with BP rose significantly. Conclusions: The resutls suggested that BP could inhibit tumor metastasis and might be a new candidate for antitumor metastasis agent.
2004, 11(1):36-38.
DOI: 10.3872/j.issn.1007-385X.2004.1.009
Abstract:
Objective: To investigate the proliferation inhibitory effect and the apoptosis-inducing activity of triptolide on HeLa cells. Methods: The effects of triptolide on cell proliferation were observed in vitro by methyl thiazolyl tetrazolium (MTT) method. The morphological alterations were confirmed by Hoechst 33258 staining. The apoptosis was evaluated by flow cytometry analysis and agarose gel electrophoresis. Results: Triptolide exhibited a marked antiproliferative effect on HeLa cells. Triptolide induced apoptosis of the cultured cells in a time- and dose-dependent manner, as evidenced by an increased percentage of annexin V/PI positive cells. Correspondingly, we observed a decrease of R123 uptake in Triptolide-treated cells, indicative of a reduction in mitochondrial membrane potential. The morphological changes of apoptotic cells and ladder pattern of DNA were observed in triptolide-treated cells. Conclusion: Triptolide can inhibit the proliferation of HeLa cells by induction of cells apoptosis.
Objective: To investigate the proliferation inhibitory effect and the apoptosis-inducing activity of triptolide on HeLa cells. Methods: The effects of triptolide on cell proliferation were observed in vitro by methyl thiazolyl tetrazolium (MTT) method. The morphological alterations were confirmed by Hoechst 33258 staining. The apoptosis was evaluated by flow cytometry analysis and agarose gel electrophoresis. Results: Triptolide exhibited a marked antiproliferative effect on HeLa cells. Triptolide induced apoptosis of the cultured cells in a time- and dose-dependent manner, as evidenced by an increased percentage of annexin V/PI positive cells. Correspondingly, we observed a decrease of R123 uptake in Triptolide-treated cells, indicative of a reduction in mitochondrial membrane potential. The morphological changes of apoptotic cells and ladder pattern of DNA were observed in triptolide-treated cells. Conclusion: Triptolide can inhibit the proliferation of HeLa cells by induction of cells apoptosis.
2004, 11(1):39-41.
DOI: 10.3872/j.issn.1007-385X.2004.1.010
Abstract:
Objective:To study the selective killing effects of immunotoxin BDI-1-PEA against human-bladder cancer cell line E-J and nude mice bearing human bladder tumor. Methods:MTT was used to measure the killing ratio of BDI-1-PEA, mixtures of BDI-1 and PEA, PEA against human-bladder cancer cell line E-J at different concentrations in vitro. When the tumor size reached 0.3 cm, BDI-1-PEA,BDI-1 and IgG were injected respectively to each group of mice though tail vein every other day for 6 times. Determined the inhibitory effects of BDI-1-PEA, BDI-1 and IgG in nude mice bearing human bladder tumor. Results: The results indicated that BDI-1-PEA showed stronger cytotoxicity against human bladder cancer cell line E-J than PEA and mixtures of BDI-1 and PEA, and stronger cytotoxicity against nude mice bearing human bladder tumor than BDI-1 and IgG.Compared with cytotoxicity against non-target cell，the difference was significant (P<0.01). Conclusion: Immunotoxin BDI-1-PEA is a largely potential anticancer drug, the result is a good foundation for the further clinical study.
Objective:To study the selective killing effects of immunotoxin BDI-1-PEA against human-bladder cancer cell line E-J and nude mice bearing human bladder tumor. Methods:MTT was used to measure the killing ratio of BDI-1-PEA, mixtures of BDI-1 and PEA, PEA against human-bladder cancer cell line E-J at different concentrations in vitro. When the tumor size reached 0.3 cm, BDI-1-PEA,BDI-1 and IgG were injected respectively to each group of mice though tail vein every other day for 6 times. Determined the inhibitory effects of BDI-1-PEA, BDI-1 and IgG in nude mice bearing human bladder tumor. Results: The results indicated that BDI-1-PEA showed stronger cytotoxicity against human bladder cancer cell line E-J than PEA and mixtures of BDI-1 and PEA, and stronger cytotoxicity against nude mice bearing human bladder tumor than BDI-1 and IgG.Compared with cytotoxicity against non-target cell，the difference was significant (P<0.01). Conclusion: Immunotoxin BDI-1-PEA is a largely potential anticancer drug, the result is a good foundation for the further clinical study.
2004, 11(1):42-45.
DOI: 10.3872/j.issn.1007-385X.2004.1.011
Abstract:
Objective:To construct attenuated salmonella typhimurium haboring an eukaryotic co-expression plasmid encoding TIP30 and human IFN-γ gene and observing its effect on the growth of adenoid cystic carcinoma. Methods:The TIP30 and human IFN-γ genes were amplified by PCR and inserted into eukaryotic expression vector pCI-neofor the construction of expression plasmids pCI-TIP30 and pCI-IFN, respectively. A co-expression plasmid pCI-TIP30/IFN was constructed by linking TIP30 and human IFN-γ gene using the sequence of internal ribosome binding sequence (IRES). Three recombinant expression plasmids were transformed into an attenuated AroA -autotrophic mutant of salmonella typhimurium SL7207, the resultant bacteria were used to infected murine macrophage in vitro and the expressed products were detected by Western blot and ELISA, respectively. Tumor growth was observed by oral administration of the recombinant salmonella typhimuriums to the nude mouse with adenoid cystic carcinoma.Results:Murine macrophage infected with recombinant salmonella transformed with both plasmids pCI-TIP30 and pCI-TIP30/IFN could express TIP30 protein, and murine macrophage infected with recombinant salmonella transformed with pCI-IFN or pCI-TIP30/IFN could secret human IFN-γ in the culture supernatant. Attenuated salmonella typhimurium and three constructed recombinant salmonella typhimuriums all had evident inhibition onthe tumor growth in nude mouse with adenoid cystic carcinoma.Conclusion: The recombinant attenuated salmonella typhimuriums haboring plasmid pCI-TIP30, plasmidpCI-IFN and co-expressing plasmidpCI-TIP30/IFN were successfully constructed, which could inhibit the growth of adenoid cystic carcinoma in nude mouse.
Objective:To construct attenuated salmonella typhimurium haboring an eukaryotic co-expression plasmid encoding TIP30 and human IFN-γ gene and observing its effect on the growth of adenoid cystic carcinoma. Methods:The TIP30 and human IFN-γ genes were amplified by PCR and inserted into eukaryotic expression vector pCI-neofor the construction of expression plasmids pCI-TIP30 and pCI-IFN, respectively. A co-expression plasmid pCI-TIP30/IFN was constructed by linking TIP30 and human IFN-γ gene using the sequence of internal ribosome binding sequence (IRES). Three recombinant expression plasmids were transformed into an attenuated AroA -autotrophic mutant of salmonella typhimurium SL7207, the resultant bacteria were used to infected murine macrophage in vitro and the expressed products were detected by Western blot and ELISA, respectively. Tumor growth was observed by oral administration of the recombinant salmonella typhimuriums to the nude mouse with adenoid cystic carcinoma.Results:Murine macrophage infected with recombinant salmonella transformed with both plasmids pCI-TIP30 and pCI-TIP30/IFN could express TIP30 protein, and murine macrophage infected with recombinant salmonella transformed with pCI-IFN or pCI-TIP30/IFN could secret human IFN-γ in the culture supernatant. Attenuated salmonella typhimurium and three constructed recombinant salmonella typhimuriums all had evident inhibition onthe tumor growth in nude mouse with adenoid cystic carcinoma.Conclusion: The recombinant attenuated salmonella typhimuriums haboring plasmid pCI-TIP30, plasmidpCI-IFN and co-expressing plasmidpCI-TIP30/IFN were successfully constructed, which could inhibit the growth of adenoid cystic carcinoma in nude mouse.
2004, 11(1):46-49.
DOI: 10.3872/j.issn.1007-385X.2004.1.012
Abstract:
Objective: To observe the transfer efficiency of protamine-DOTAP/Chol liposome-oligodeoxynuleotides complex to SKOV3 cells. Methods: Synthesized NF-κB decoy ODNs or scrambled ODNs labeled by FITC on the 5′ends was mixed with rationic DOTAP/cholesteral liposome and protamine, making the charge ratio of positive:negative 4. SKOV3 cells were transfected by the complex(1 μmol/L) and the efficiency were measured by flow cytometry(FCM) and fluorescence meter. The direct effect of NF-κB decoy ODNs transfer on the growth of SKOV3 cell line in vitro was measured by MTT method. Results: The efficiency of LPD transferto SKOV3 cell line was 96.41%, significantly higher than that of naked DNA transfer which was 12.86%.The growth of SKOV3 cell line was not affected by 48h incubation after NF-κB decoy ODNs or scrambled ODNs transfection.Conclusion: LPD method is the one with high transfer efficiency not affected by NBS(newborn bovine serum)with possibility of use in viv.
Objective: To observe the transfer efficiency of protamine-DOTAP/Chol liposome-oligodeoxynuleotides complex to SKOV3 cells. Methods: Synthesized NF-κB decoy ODNs or scrambled ODNs labeled by FITC on the 5′ends was mixed with rationic DOTAP/cholesteral liposome and protamine, making the charge ratio of positive:negative 4. SKOV3 cells were transfected by the complex(1 μmol/L) and the efficiency were measured by flow cytometry(FCM) and fluorescence meter. The direct effect of NF-κB decoy ODNs transfer on the growth of SKOV3 cell line in vitro was measured by MTT method. Results: The efficiency of LPD transferto SKOV3 cell line was 96.41%, significantly higher than that of naked DNA transfer which was 12.86%.The growth of SKOV3 cell line was not affected by 48h incubation after NF-κB decoy ODNs or scrambled ODNs transfection.Conclusion: LPD method is the one with high transfer efficiency not affected by NBS(newborn bovine serum)with possibility of use in viv.
12 Construction of Prokarytoic Expression Vector of TRAIL Gene and Preparation of Anti-TRAIL Antibody
FANG Qin
,
SUN Deng-jun
,
ZHANG Xu
,
REN Zhi-juan
,
XIAO Yun
,
LIU Feng-feng
,
WU Zheng-yu
,
WEN Xiao-ping
,
WANG Ji-shi
2004, 11(1):50-53.
DOI: 10.3872/j.issn.1007-385X.2004.1.013
Abstract:
Objective:To obtain the anti-TRAIL antibody. Methods:A prokaryotic expression vector pPre-TMHTb was constructed by PCR and recombinant DNA techniques and was confirmed by enzymatic digestion and sequence analysis. TRAIL protein induced to express by IPTG in E.coli DH-5α. After TRAIL protein immunised rabbits, anti-TRAIL antibody was tested by Dot blot and Western blot. Results:A prokaryotic expression vector pPre-TMHTb was constructed and expressd in E.coli DH-5α. TRAIL protein was detected up to 11.8% of the total bacterial protein expressed and anti-TRAIL antibody was tested by Dot blot and Western blot. Conclusions: The anti-TRAIL antibody was obtained that provides an experimental basis for the studies of TRAIL in eukaryotic level.
Objective:To obtain the anti-TRAIL antibody. Methods:A prokaryotic expression vector pPre-TMHTb was constructed by PCR and recombinant DNA techniques and was confirmed by enzymatic digestion and sequence analysis. TRAIL protein induced to express by IPTG in E.coli DH-5α. After TRAIL protein immunised rabbits, anti-TRAIL antibody was tested by Dot blot and Western blot. Results:A prokaryotic expression vector pPre-TMHTb was constructed and expressd in E.coli DH-5α. TRAIL protein was detected up to 11.8% of the total bacterial protein expressed and anti-TRAIL antibody was tested by Dot blot and Western blot. Conclusions: The anti-TRAIL antibody was obtained that provides an experimental basis for the studies of TRAIL in eukaryotic level.
2004, 11(1):54-55.
DOI: 10.3872/j.issn.1007-385X.2004.1.014
Abstract:
Objective: To investigate the proliferation inhibitory effect and the apoptosis-inducing activity of triptolide on HeLa cells. Methods: The effects of triptolide on cell proliferation were observed in vitro by methyl ihiazolyl tetrazoli-um ( MTT) method. The morphological alterations were confirmed by Hoechst 33258 staining. The apoptosis was evaluated by flow cytometry analysis and agarose gel electrophoresis. Results: Triptolide exhibited a marked antiproliferative effect on HeLa cells. Triptolide induced apoptosis of the cultured cells in a time- and dose-dependent manner, as evidenced by an increased percentage of annexin V/PI positive cells. Correspondingly, we observed a decrease of R123 uptake in Triptolide-treated cells, indicative of a reduction in mitochondrial membrane potential. The morphological changes of apoptotic cells and ladder pattern of DNA were observed in triptolide-treated cells. Conclusion: Triptolide can inhibit the proliferation of HeLa cells by induction of cells apoptosis.
Objective: To investigate the proliferation inhibitory effect and the apoptosis-inducing activity of triptolide on HeLa cells. Methods: The effects of triptolide on cell proliferation were observed in vitro by methyl ihiazolyl tetrazoli-um ( MTT) method. The morphological alterations were confirmed by Hoechst 33258 staining. The apoptosis was evaluated by flow cytometry analysis and agarose gel electrophoresis. Results: Triptolide exhibited a marked antiproliferative effect on HeLa cells. Triptolide induced apoptosis of the cultured cells in a time- and dose-dependent manner, as evidenced by an increased percentage of annexin V/PI positive cells. Correspondingly, we observed a decrease of R123 uptake in Triptolide-treated cells, indicative of a reduction in mitochondrial membrane potential. The morphological changes of apoptotic cells and ladder pattern of DNA were observed in triptolide-treated cells. Conclusion: Triptolide can inhibit the proliferation of HeLa cells by induction of cells apoptosis.
2004, 11(1):55-57.
DOI: 10.3872/j.issn.1007-385X.2004.1.015
Abstract:
Objective: To study the selective killing effects of immunotoxin BDI-1-PEA against human-bladder cancer cell line E-J and nude mice bearing human bladder tumor. Methods; MTT was used to measure the killing ratio of BDI-1-PEA, mixtures of BDI-1 and PEA, PEA against human-bladder cancer cell line E-J at different concentrations in vitro. When the tumor size reached 0. 3 cm, BDI-1-PEA,BDI-1 and IgG were injected respectively to each group of mice though tail vein every other day for 6 times. Determined the inhibitory effects of BDI-1-PEA, BDI-1 and IgG in nude mice bearing human bladder tumor. Results; The results indicated that BDI-1 -PEA showed stronger cytotoxicity against human bladder cancer cell line E-J than PEA and mixtures of BDI-1 and PEA, and stronger cytotoxicity against nude mice bearing human bladder tumor than BDI-1 and IgG. Compared with cytotoxicity against non-target cell, the difference was significant ( P < 0.01). Conclusion: Immunotoxin BDI-1-PEA is a largely potential anticancer drug, the result is a good foundation for the further clinical study.
Objective: To study the selective killing effects of immunotoxin BDI-1-PEA against human-bladder cancer cell line E-J and nude mice bearing human bladder tumor. Methods; MTT was used to measure the killing ratio of BDI-1-PEA, mixtures of BDI-1 and PEA, PEA against human-bladder cancer cell line E-J at different concentrations in vitro. When the tumor size reached 0. 3 cm, BDI-1-PEA,BDI-1 and IgG were injected respectively to each group of mice though tail vein every other day for 6 times. Determined the inhibitory effects of BDI-1-PEA, BDI-1 and IgG in nude mice bearing human bladder tumor. Results; The results indicated that BDI-1 -PEA showed stronger cytotoxicity against human bladder cancer cell line E-J than PEA and mixtures of BDI-1 and PEA, and stronger cytotoxicity against nude mice bearing human bladder tumor than BDI-1 and IgG. Compared with cytotoxicity against non-target cell, the difference was significant ( P < 0.01). Conclusion: Immunotoxin BDI-1-PEA is a largely potential anticancer drug, the result is a good foundation for the further clinical study.
2004, 11(1):64-66.
DOI: 10.3872/j.issn.1007-385X.2004.1.018
Abstract:
美罗华单克隆抗体具有高度特异性,毒副作用小,是近年来淋巴瘤治疗上的突破性进展。临床可用于各种B细胞NHL的一线或二线治疗，可单独应用，亦可与化疗药物联合应用以提高疗效。Zevalin为90Y 标记的鼠抗CD20单抗，于2002年通过FDA批准。可用于治疗美罗华治疗无效或复发的NHL患者，有效率仍可高达54％。
美罗华单克隆抗体具有高度特异性,毒副作用小,是近年来淋巴瘤治疗上的突破性进展。临床可用于各种B细胞NHL的一线或二线治疗，可单独应用，亦可与化疗药物联合应用以提高疗效。Zevalin为90Y 标记的鼠抗CD20单抗，于2002年通过FDA批准。可用于治疗美罗华治疗无效或复发的NHL患者，有效率仍可高达54％。
16 Construction of Prokarytoic Expression Vector of TRAIL Gene and Preparation of Anti-TRAIL Antibody
2004, 11(1):70-72.
DOI: 10.3872/j.issn.1007-385X.2004.1.020
Abstract:
RNA类小分子可作为治疗性药物应用于各类疾病的治疗。这类小分子主要包括基因表达抑制剂、基因修复剂、蛋白拮抗剂及RNA疫苗，目前已有多种反义RNA和核酶作为基因表达抑制剂正在进行各期临床试验，它们主要用于心血管疾病、肿瘤、药物代谢性疾病、病毒性肝炎和艾滋病等疾病的治疗。蛋白拮抗剂aptamers已作为治疗老年性黄斑和血栓药物进入临床Ⅱ,Ⅲ期试验。RNA疫苗作为免疫调节剂治疗前列腺癌和肾癌的临床试验也在进行中。
RNA类小分子可作为治疗性药物应用于各类疾病的治疗。这类小分子主要包括基因表达抑制剂、基因修复剂、蛋白拮抗剂及RNA疫苗，目前已有多种反义RNA和核酶作为基因表达抑制剂正在进行各期临床试验，它们主要用于心血管疾病、肿瘤、药物代谢性疾病、病毒性肝炎和艾滋病等疾病的治疗。蛋白拮抗剂aptamers已作为治疗老年性黄斑和血栓药物进入临床Ⅱ,Ⅲ期试验。RNA疫苗作为免疫调节剂治疗前列腺癌和肾癌的临床试验也在进行中。
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- 《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
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- 网址:http://www.biother.cn
- 刊号：ISSN 1007-385X
- CN 31-1725/R
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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.