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Volume 9,Issue 4,2002 Table of Contents
1 Combined Interleukin 12 and Interleukin 2 Gene Therapy for Hepatocellular Carcinoma in A Rat Model
YANG Jia-he
,
FAN Rui-fang
,
QIAN Qi-jun
,
YOU Tian-geng
,
XUE Hui-bin
,
SU Chang-qing
,
CAO Hui-fang
,
WU Meng-chao
2002, 9(4):233-236.
DOI: 10.3872/j.issn.1007-385X.2002.4.002
Abstract:
Objective:To determine the feasibility and efficacy of combined murine interleukin 12 (mIL-12) and human interleukin 2 (hIL-2) gene therapy for hepatocellular carcinoma in a rat model. Methods: The retroviral vector encoding mIL-12/hIL-2 gene was constructed and then transfected into packaging cell line. The cells were injected into rats in the established hepatoma at different time points. The therapeutic effect, immune function and toxicological response were evaluated. Results:Intratumoral injection of recombinant retroviral vector encoding mIL-12/hIL-2 gene resulted in marked hepatoma regression. The 35 d survival rates of the subgroups treated on the first，third，fifth and seventh day after tumor implantation were 100%, 100%, 30% and 10% respectively. The average survival time of the IL-12+IL-2 treatment group was superior to those of the physiological saline group（P＜0.01), retroviral empty vector group（P＜0.01), IL-2 treatment group(P＜0.01) and IL-12 treatment group（P＜0.01). The immunological study showed that the number of hepatoma infiltrating lymphocytes was increased in the IL-12+IL-2 treatment group. Conclusion:The retroviral packaging cell line encoding mIL-12 and hIL-2 gene via intratumoral injection inhibits the growth of hepatocellular carcinoma significantly. The therapeutical effects of early administration is superior to that of later one.
Objective:To determine the feasibility and efficacy of combined murine interleukin 12 (mIL-12) and human interleukin 2 (hIL-2) gene therapy for hepatocellular carcinoma in a rat model. Methods: The retroviral vector encoding mIL-12/hIL-2 gene was constructed and then transfected into packaging cell line. The cells were injected into rats in the established hepatoma at different time points. The therapeutic effect, immune function and toxicological response were evaluated. Results:Intratumoral injection of recombinant retroviral vector encoding mIL-12/hIL-2 gene resulted in marked hepatoma regression. The 35 d survival rates of the subgroups treated on the first，third，fifth and seventh day after tumor implantation were 100%, 100%, 30% and 10% respectively. The average survival time of the IL-12+IL-2 treatment group was superior to those of the physiological saline group（P＜0.01), retroviral empty vector group（P＜0.01), IL-2 treatment group(P＜0.01) and IL-12 treatment group（P＜0.01). The immunological study showed that the number of hepatoma infiltrating lymphocytes was increased in the IL-12+IL-2 treatment group. Conclusion:The retroviral packaging cell line encoding mIL-12 and hIL-2 gene via intratumoral injection inhibits the growth of hepatocellular carcinoma significantly. The therapeutical effects of early administration is superior to that of later one.
WANG Qian
,
CAO Li-min
,
ZHOU Hua-rong
,
ZHU Hui-fen
,
ZHANG Yue
,
SHAO Jin-fang
,
YANG Jing
,
SHEN Guan-xin
2002, 9(4):243-247.
DOI: 10.3872/j.issn.1007-385X.2002.4.004
Abstract:
Objective: To construct the potent targeting gene delivery and expression system, and to investigate the special killing effect of HSV-tk/GCV system on human liver cancer cells in vitro mediated by it. Methods: The anti-TfR ScFv-GAL4 fusion protein expression vector ScFv-GAL4-pET28a and the eukaryocyte expression plasmid pEBAF/tk-GAL4rec were constructed by recombinant DNA technology. After the induction by IPTG, we got the anti-TfR ScFv-GAL4 fusion protein as delivery vector to transfect pEBAF/tk-GAL4rec into the human liver cancer cells line HepG2,SMMC7721 and lung cancer cells line A549 which all over-express TfR via receptor-mediated endocytosis. The positive cell clones were selected by hygromycin and were named HepG2/tk,SMMC7721/tk and A549/tk respectively. Then MTT method was used to determine the killing effect of GCV on them. Results: The constructions of the ScFv-Gal4 fusion protein and the recombination expression plasmid pEBAF/tk-GAL4rec were confirmed by double enzyme digestion, SDS-PAGE and sequencing. In the experiment of cytotoxicity effect, the HepG2/tk cells that over-secrete AFP (845 ng/ml) were highly sensitive to the toxicity of GCV, whereas the SMMC7721/tk cells that under-secrete AFP (2 ng/ml) were slightly sensitive to GCV, and the A549/tk cells that don t secrete AFP were not.Conclusions:The double-targeting and tissue-specific HSV-tk/GCV anti-tumor system has been constructed successfully, and it displays a good targeting to tumor cells.
Objective: To construct the potent targeting gene delivery and expression system, and to investigate the special killing effect of HSV-tk/GCV system on human liver cancer cells in vitro mediated by it. Methods: The anti-TfR ScFv-GAL4 fusion protein expression vector ScFv-GAL4-pET28a and the eukaryocyte expression plasmid pEBAF/tk-GAL4rec were constructed by recombinant DNA technology. After the induction by IPTG, we got the anti-TfR ScFv-GAL4 fusion protein as delivery vector to transfect pEBAF/tk-GAL4rec into the human liver cancer cells line HepG2,SMMC7721 and lung cancer cells line A549 which all over-express TfR via receptor-mediated endocytosis. The positive cell clones were selected by hygromycin and were named HepG2/tk,SMMC7721/tk and A549/tk respectively. Then MTT method was used to determine the killing effect of GCV on them. Results: The constructions of the ScFv-Gal4 fusion protein and the recombination expression plasmid pEBAF/tk-GAL4rec were confirmed by double enzyme digestion, SDS-PAGE and sequencing. In the experiment of cytotoxicity effect, the HepG2/tk cells that over-secrete AFP (845 ng/ml) were highly sensitive to the toxicity of GCV, whereas the SMMC7721/tk cells that under-secrete AFP (2 ng/ml) were slightly sensitive to GCV, and the A549/tk cells that don t secrete AFP were not.Conclusions:The double-targeting and tissue-specific HSV-tk/GCV anti-tumor system has been constructed successfully, and it displays a good targeting to tumor cells.
2002, 9(4):248-252.
DOI: 10.3872/j.issn.1007-385X.2002.4.005
Abstract:
Objective: Designing dendritic-cell-based vaccines against cancer concerned with the infection of EBV to solve tumor escape. Methods: CD34+ hematopoietic stem cells of bone marrow were cultured in presence of hGM-CSF,hTNF-α,hIL-3,hIL-4 in vitro for two weeks to obtain large amount of DC. DC were characterized by FACS analysis and pulsed with the EBV envelope glycoprotein gp340. The gp340-loaded DC initiate T lymphocytes to induce T lymphocytes activated (CTL) against cancer concerned with the infection of EBV. Results: Marrow-derived DC expressing high levels of CD1а have typical dendritic morphology. They could acquire Epstein-Barr virus latent membrane glycoprotein gp340 efficiently and induce T cells increasing stimulatory capacity in MLR. Only gp340-loaded DC could induce special CTL against tumor cells concerned with the infection of EBV in cytotoxicity assays. Conclusion: Using DC pulsed with EBV vaccines against EBV infection could start up anti-tumor immunity and induce specific CTLs to availability kill tumor cells associated with EBV in vitro.
Objective: Designing dendritic-cell-based vaccines against cancer concerned with the infection of EBV to solve tumor escape. Methods: CD34+ hematopoietic stem cells of bone marrow were cultured in presence of hGM-CSF,hTNF-α,hIL-3,hIL-4 in vitro for two weeks to obtain large amount of DC. DC were characterized by FACS analysis and pulsed with the EBV envelope glycoprotein gp340. The gp340-loaded DC initiate T lymphocytes to induce T lymphocytes activated (CTL) against cancer concerned with the infection of EBV. Results: Marrow-derived DC expressing high levels of CD1а have typical dendritic morphology. They could acquire Epstein-Barr virus latent membrane glycoprotein gp340 efficiently and induce T cells increasing stimulatory capacity in MLR. Only gp340-loaded DC could induce special CTL against tumor cells concerned with the infection of EBV in cytotoxicity assays. Conclusion: Using DC pulsed with EBV vaccines against EBV infection could start up anti-tumor immunity and induce specific CTLs to availability kill tumor cells associated with EBV in vitro.
ZHANG Li-hong
,
JIA Lin-tao
,
CHEN Guang-sheng
,
QU Ping
,
YU Cui-juan
,
ZHAO Jing
,
XU Yan-ming
,
WANG Cheng-ji
,
YANG An-gang
2002, 9(4):253-256.
DOI: 10.3872/j.issn.1007-385X.2002.4.006
Abstract:
Objective: To investigate the apoptosis induction effect due to the expression of reversed caspase-3 gene in SKBr3 cells.Methods: Reversed caspase-3 gene was gain by recombinant PCR and cloned into expression vector pcDNA3 to transfected SKBr3 cells. The expression and the apoptosis induction effect of reversed caspase-3 gene on SKBr3 cells were observed by HE and immunohistochemical staining. The cell cycle of transfected SKBr3 cells were analyzed by FCM. The tumor suppression effect of reversed caspase-3 gene was observed through administrating recombinant plasmids to BALB/c nude mouses bearing SKBr3 tumor.Results: Reversed caspase-3 gene can be expressed in SKBr3 stably. Reversed caspased-3 protein can induce obvious apoptosis in vitro and inhibit tumor growth in vivo.Conclusion: Reversed caspase-3 gene can effectively induce SKBr3 cells to apoptosis.
Objective: To investigate the apoptosis induction effect due to the expression of reversed caspase-3 gene in SKBr3 cells.Methods: Reversed caspase-3 gene was gain by recombinant PCR and cloned into expression vector pcDNA3 to transfected SKBr3 cells. The expression and the apoptosis induction effect of reversed caspase-3 gene on SKBr3 cells were observed by HE and immunohistochemical staining. The cell cycle of transfected SKBr3 cells were analyzed by FCM. The tumor suppression effect of reversed caspase-3 gene was observed through administrating recombinant plasmids to BALB/c nude mouses bearing SKBr3 tumor.Results: Reversed caspase-3 gene can be expressed in SKBr3 stably. Reversed caspased-3 protein can induce obvious apoptosis in vitro and inhibit tumor growth in vivo.Conclusion: Reversed caspase-3 gene can effectively induce SKBr3 cells to apoptosis.
2002, 9(4):257-260.
DOI: 10.3872/j.issn.1007-385X.2002.4.007
Abstract:
Objective：To construct a humanized fusion protein consisting of a EGF linked to a human angiogenin(Ang) and evaluate the potential of the construct(EGF-Ang) to target and kill tumor cells.Methods：By using of genetic engineering techniques, we constructed a recombinant expressing plasmid pEGF-Ang, and expressed fusion protein accumulated in intracellular inclusion bodies. The recombinant EGF-Ang proteins were purified through DEAE-Sepharose FF chromatography in the denatured condition. After renaturing process, the cytotoxicity was measured with MTT colorimetric assay in vitro. Results：A novel fusion protein named EGF-Ang was constructed and expressed in E coli. Analysis of SDS-PAGE and thin layer scanning results showed that amount of expressed fusion protein was 18.6% in total lysis protein of bacteria. Cytotoxicity analysis showed that the fusion proteins could inhibit the growth of Hep2 cells but had little influence on the growth of MA104 cells. Conclusion：EGF-Ang fusion protein had definitely cytotoxicity to Hep2 cells which over expressed EGFR in vitro.
Objective：To construct a humanized fusion protein consisting of a EGF linked to a human angiogenin(Ang) and evaluate the potential of the construct(EGF-Ang) to target and kill tumor cells.Methods：By using of genetic engineering techniques, we constructed a recombinant expressing plasmid pEGF-Ang, and expressed fusion protein accumulated in intracellular inclusion bodies. The recombinant EGF-Ang proteins were purified through DEAE-Sepharose FF chromatography in the denatured condition. After renaturing process, the cytotoxicity was measured with MTT colorimetric assay in vitro. Results：A novel fusion protein named EGF-Ang was constructed and expressed in E coli. Analysis of SDS-PAGE and thin layer scanning results showed that amount of expressed fusion protein was 18.6% in total lysis protein of bacteria. Cytotoxicity analysis showed that the fusion proteins could inhibit the growth of Hep2 cells but had little influence on the growth of MA104 cells. Conclusion：EGF-Ang fusion protein had definitely cytotoxicity to Hep2 cells which over expressed EGFR in vitro.
2002, 9(4):261-264.
DOI: 10.3872/j.issn.1007-385X.2002.4.008
Abstract:
Objective:To explore caspase-6′s effect on the apoptosis of osteosarcoma cell line SOSP-9607. Methods:The expression level of caspase-6 in the osteosarcoma cell line SOSP-9067 was examined by RT-PCR method. The adenovirus adv5 vector with caspase-6 gene was constructed and transferred into the osteosarcoma cell line SOSP-9607 by lipofection. The cell survival rate after transfection was assayed by MTT method. The cell morphological changes were observed by microscope and electron microscope, the apoptosis of transferred cells were examined by gel electrophoresis. Results: No expression of caspase-6 was examined in the osteosarcoma cell line SOSP-9607. A high expression of caspase-6 was identified by RT-PCR after the transfection. The cell growth curve declined after transferring caspase-6. Electrophoresis of DNA displayed the apoptosis ladder.Conclusion:Transferring caspase-6 into the osteosarcoma line SOSP-9607 may inhibit the growth of the osteosarcoma cell line SOSP-9607 and this effect might be achieved by inducing apoptosis.
Objective:To explore caspase-6′s effect on the apoptosis of osteosarcoma cell line SOSP-9607. Methods:The expression level of caspase-6 in the osteosarcoma cell line SOSP-9067 was examined by RT-PCR method. The adenovirus adv5 vector with caspase-6 gene was constructed and transferred into the osteosarcoma cell line SOSP-9607 by lipofection. The cell survival rate after transfection was assayed by MTT method. The cell morphological changes were observed by microscope and electron microscope, the apoptosis of transferred cells were examined by gel electrophoresis. Results: No expression of caspase-6 was examined in the osteosarcoma cell line SOSP-9607. A high expression of caspase-6 was identified by RT-PCR after the transfection. The cell growth curve declined after transferring caspase-6. Electrophoresis of DNA displayed the apoptosis ladder.Conclusion:Transferring caspase-6 into the osteosarcoma line SOSP-9607 may inhibit the growth of the osteosarcoma cell line SOSP-9607 and this effect might be achieved by inducing apoptosis.
MA Weng-xiong
,
WANG Weng-hong
,
CHENG Gui-ling
,
HUI Guo-zheng
,
WU Gie
,
ZHANG Shi-ming
,
ZHOU Dai
,
DU Zi-wei
2002, 9(4):265-268.
DOI: 10.3872/j.issn.1007-385X.2002.4.009
Abstract:
Objective: To evaluate effects of Fas ligand (FasL) gene transfer in vitro for apoptosis of malignant melanoma. Methods: We constructed recombinant adenoviral vector containing human FasL cDNA(Ad-FL) under the control of a cytomegalovirus (CMV) promoter, which encodes human Fas ligand and transfected into two melanoma cells. Flow cytometric analysis, RT-PCR identify the expression of Fas/FasL. TUNEL, fluorescence microscope were used to analyse apoptosis. Results: Two melanoma cells surface express Fas, whereas not express FasL detected by RT-PCR and flow cytometric analysis; while tumor cells transducted by Ad-FL can express high level of FasL. Ad-FL can induce two melanoma cells apoptosis or suppress their growth in vitro. Conclusion: Recombinant adenovirus FasL had great effect on inducing the apoptosis of human melanoma in vitro.
Objective: To evaluate effects of Fas ligand (FasL) gene transfer in vitro for apoptosis of malignant melanoma. Methods: We constructed recombinant adenoviral vector containing human FasL cDNA(Ad-FL) under the control of a cytomegalovirus (CMV) promoter, which encodes human Fas ligand and transfected into two melanoma cells. Flow cytometric analysis, RT-PCR identify the expression of Fas/FasL. TUNEL, fluorescence microscope were used to analyse apoptosis. Results: Two melanoma cells surface express Fas, whereas not express FasL detected by RT-PCR and flow cytometric analysis; while tumor cells transducted by Ad-FL can express high level of FasL. Ad-FL can induce two melanoma cells apoptosis or suppress their growth in vitro. Conclusion: Recombinant adenovirus FasL had great effect on inducing the apoptosis of human melanoma in vitro.
2002, 9(4):269-271.
DOI: 10.3872/j.issn.1007-385X.2002.4.010
Abstract:
Objective： Tiam-1 antisense gene was used to downregulate Tiam-1 mRNA and protein expression, thus tumor metastasis could be suppressed. Methods：In this study Tiam-1 mRNA was examined by RT-PCR. The expression of Rho protein was examined by flow cytometry. Basement membrane was used to examine the effect of Tiam 1 antisense gene on metastatic behavior of PG cells. Results：The relative expression level of Tiam-1 mRNA was reduced after exposure to Tiam-1 ASODNS-Lf. Only 72 hours of exposure to Tiam-1 ASODNS, cell metastasis of treated group was inhibitted significantly. The invasive cell numbers through matrigel reduced from 945±40 to 649±35 (P<005). Tiam-1 ASODNS-Lf significantly inhibitted Rho protein expression ratio from 27.86% to 16.86%(P<0.001), which related to signal transduction pathway. Conclusions：Antisense gene inhibits invasion and metastasis of tumor cells through a cascade reaction of gene-protein-cell effect and provides the theoretical and experimental bases for clinical therapy and study of antimetastasis.
Objective： Tiam-1 antisense gene was used to downregulate Tiam-1 mRNA and protein expression, thus tumor metastasis could be suppressed. Methods：In this study Tiam-1 mRNA was examined by RT-PCR. The expression of Rho protein was examined by flow cytometry. Basement membrane was used to examine the effect of Tiam 1 antisense gene on metastatic behavior of PG cells. Results：The relative expression level of Tiam-1 mRNA was reduced after exposure to Tiam-1 ASODNS-Lf. Only 72 hours of exposure to Tiam-1 ASODNS, cell metastasis of treated group was inhibitted significantly. The invasive cell numbers through matrigel reduced from 945±40 to 649±35 (P<005). Tiam-1 ASODNS-Lf significantly inhibitted Rho protein expression ratio from 27.86% to 16.86%(P<0.001), which related to signal transduction pathway. Conclusions：Antisense gene inhibits invasion and metastasis of tumor cells through a cascade reaction of gene-protein-cell effect and provides the theoretical and experimental bases for clinical therapy and study of antimetastasis.
2002, 9(4):272-275.
DOI: 10.3872/j.issn.1007-385X.2002.4.011
Abstract:
Objective: To explore the mechanism and reversion of malignant phenotypes in a highly metastatic human lung tumor cell line. Methods: Suppression of ICA and PJM on PG cell line and the effects of PJM and ICA on PG cells sensitivity to lysis by CD3AK effectors cell was studied by MTT assay; the expression of bcl-2 and c-fos was studied by RT-PCR and flow cytometry, the expression of HLA-A,B,C in PG cells was examined by flow cytometry. Results: ICA and PJM inhibited the growth of PG obviously. ICA and PJM could significantly enhance the expression of HLA-ABC antigen and c-fos gene. ICA and PJM decreased the expression of bcl-2 and enhanced the the susceptibility of PG cells to lysis by CD3AK cells.Conclusion: ICA and PJM can reverse the malignant phenotypes of PG cells, enhance the recognition and killing effect of T cells.
Objective: To explore the mechanism and reversion of malignant phenotypes in a highly metastatic human lung tumor cell line. Methods: Suppression of ICA and PJM on PG cell line and the effects of PJM and ICA on PG cells sensitivity to lysis by CD3AK effectors cell was studied by MTT assay; the expression of bcl-2 and c-fos was studied by RT-PCR and flow cytometry, the expression of HLA-A,B,C in PG cells was examined by flow cytometry. Results: ICA and PJM inhibited the growth of PG obviously. ICA and PJM could significantly enhance the expression of HLA-ABC antigen and c-fos gene. ICA and PJM decreased the expression of bcl-2 and enhanced the the susceptibility of PG cells to lysis by CD3AK cells.Conclusion: ICA and PJM can reverse the malignant phenotypes of PG cells, enhance the recognition and killing effect of T cells.
2002, 9(4):276-279.
DOI: 10.3872/j.issn.1007-385X.2002.4.012
Abstract:
Objective: To investigate the inhibitory effects of human angiostatin (AS) on the subcutaneously transplanted mouse glioma. Methods: Effects of AS on the proliferation of angioendothelial cell line ECV-304 and human glioma cell line TJ-905 were observed with MTT method. Mouse model of subcutaneously transplanted of glioma line G422 was established. AS was injected subcutaneously, tumor ratio and tumor in hibitory rate were estimated. Results: (1) Inhibitory effects of AS on ECV-304 enhanced with the increase of dose, while AS exerted no effect on TJ-905 ; (2) It was shown with animal experiments that tumer inhibitory rates were 21.8% and 84.3% with AS doses of 10 mg and 50 mg·kg-1·d-1, respectively; (3) Immunochemical factor Ⅷ staining indicated there the differences between the control and the control and the test groups in both the angiodevelopment and quantity. Conclusion: There was no inhibitory effect for AS to angiolorna cell in vitro, however in vivo, it inhibited the glioma growth via its inhibitory effect on the proliferation of angioend othelial cell line. It was then suggested that AS would be of wide application in tumor treatment.
Objective: To investigate the inhibitory effects of human angiostatin (AS) on the subcutaneously transplanted mouse glioma. Methods: Effects of AS on the proliferation of angioendothelial cell line ECV-304 and human glioma cell line TJ-905 were observed with MTT method. Mouse model of subcutaneously transplanted of glioma line G422 was established. AS was injected subcutaneously, tumor ratio and tumor in hibitory rate were estimated. Results: (1) Inhibitory effects of AS on ECV-304 enhanced with the increase of dose, while AS exerted no effect on TJ-905 ; (2) It was shown with animal experiments that tumer inhibitory rates were 21.8% and 84.3% with AS doses of 10 mg and 50 mg·kg-1·d-1, respectively; (3) Immunochemical factor Ⅷ staining indicated there the differences between the control and the control and the test groups in both the angiodevelopment and quantity. Conclusion: There was no inhibitory effect for AS to angiolorna cell in vitro, however in vivo, it inhibited the glioma growth via its inhibitory effect on the proliferation of angioend othelial cell line. It was then suggested that AS would be of wide application in tumor treatment.
XIE Li-yu
,
YANG Jue
,
HAO Qing-ling
,
ZHAO Wei
,
PANG Rui-ling
,
MA Zhi
,
HUANG Jian-qiang
,
DONG Xue-xian
,
YAO Fa-xian
2002, 9(4):280-282.
DOI: 10.3872/j.issn.1007-385X.2002.4.013
Abstract:
Objective：To evaluate the value of lung cancer treatment by chemotherapeutic drugs through bronchoartery perfusion combined with interferon.Methods：40 cases with lung cancer were randomly divided into 2 groups. 20 cases accepted chemotherapeutic drugs through bronchoartery perfusion combined with interferon were considered as observed group, and the other 20 cases accepted only chemotherapeutic drugs through bronchoartery perfusion as control group. Χ2 test was used to compare the differences of the recent curative effects and survival rates between the two groups. Results：The total effective rate (CR+PR) of the observed group during the first treatment was 60.0% (12/20), which was not significant as compared with that of control group (40.0%) (Χ2= 1.60, P＞0.05). But the survival rates of the observed group during 1 and 2 years were 85.0% (17/20) and 45.0% (9/20), respectively. There were significant differences as compared with 50.0% (10/20) and 10.0% (2/20) in control group (Χ2= 5.58 and 6.14, P＜0.05).Conclusion：Bronchoartery perfusion with chemotherapeutic drugs combined with interferon can raise the survival rates of patients with lung cancer, and is a valuably therapeutic method.
Objective：To evaluate the value of lung cancer treatment by chemotherapeutic drugs through bronchoartery perfusion combined with interferon.Methods：40 cases with lung cancer were randomly divided into 2 groups. 20 cases accepted chemotherapeutic drugs through bronchoartery perfusion combined with interferon were considered as observed group, and the other 20 cases accepted only chemotherapeutic drugs through bronchoartery perfusion as control group. Χ2 test was used to compare the differences of the recent curative effects and survival rates between the two groups. Results：The total effective rate (CR+PR) of the observed group during the first treatment was 60.0% (12/20), which was not significant as compared with that of control group (40.0%) (Χ2= 1.60, P＞0.05). But the survival rates of the observed group during 1 and 2 years were 85.0% (17/20) and 45.0% (9/20), respectively. There were significant differences as compared with 50.0% (10/20) and 10.0% (2/20) in control group (Χ2= 5.58 and 6.14, P＜0.05).Conclusion：Bronchoartery perfusion with chemotherapeutic drugs combined with interferon can raise the survival rates of patients with lung cancer, and is a valuably therapeutic method.
JIANG Ri-cheng
,
FANG Wei-yi
,
DONG Bi-hua
,
DONG Lin
,
TANG Yun-lian
,
PENG Shu-ping
,
ZHOU Jian-guo
,
CAO Jian-guo
2002, 9(4):283-286.
DOI: 10.3872/j.issn.1007-385X.2002.4.014
Abstract:
Objective: To clone and to express human canstatin gene and investigate biological activity of the recombinant protein.Methods: The cDNA of canstatin was amplified with RT-PCR from fresh fetal liver and was cloned into pMD18-T vector , and was sequenced.Then canstatin cDNA was cloned into the BamHⅠand HindⅢ sites of pQE30 and expressed with induction of IPTG. After the purification under native conditions, The biological activity of the recombinant protein was identified by the chick embryo chorioallantoic membrane assay (CAM). Results:The sequence of the amplified DNA fragment is consistent with that of the known gene. The recombinant protein was highly expressed after induction with IPTG. Biological assay results indicate that the recombinant canstatin protein could suppress the new blood vessel formation in CAM in vitro.Conclusion: The cloning and expression vector of canstatin cDNA has been constructed successfully. The recombinant canstatin protein was highly expressed in E.coli M15. In the CAM, the recombinant protein has highly suppressive effects on the vessels in chick embryo chorioallantoic membrane.
Objective: To clone and to express human canstatin gene and investigate biological activity of the recombinant protein.Methods: The cDNA of canstatin was amplified with RT-PCR from fresh fetal liver and was cloned into pMD18-T vector , and was sequenced.Then canstatin cDNA was cloned into the BamHⅠand HindⅢ sites of pQE30 and expressed with induction of IPTG. After the purification under native conditions, The biological activity of the recombinant protein was identified by the chick embryo chorioallantoic membrane assay (CAM). Results:The sequence of the amplified DNA fragment is consistent with that of the known gene. The recombinant protein was highly expressed after induction with IPTG. Biological assay results indicate that the recombinant canstatin protein could suppress the new blood vessel formation in CAM in vitro.Conclusion: The cloning and expression vector of canstatin cDNA has been constructed successfully. The recombinant canstatin protein was highly expressed in E.coli M15. In the CAM, the recombinant protein has highly suppressive effects on the vessels in chick embryo chorioallantoic membrane.
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Chinese Journal Of Cancer Biotheray ® 2025 All Rights Reserved
Supported by:Beijing E-Tiller Technology Development Co., Ltd.