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Volume 7,Issue 2,2000 Table of Contents
2000, 7(2):83-85.
DOI: 10.3872/j.issn.1007-385X.2000.2.002
Abstract:
To investigate which step of IL-6 signal transduction pathways in myeloma cells can be taken as the acting target of the antagonists for IL-6. Methods: EMSA and immunoprecipitation were used to detect the activation of transcription factors(TFs)-STAT3，NF-IL-6 and protein kinase ERK in a myeloma cell line-Sko-007 by IL-6, then anti-sense expression plasmids and anti-sense ODN for these signal moleculars were constructed and designed,the effects of these anti-sense nucleic acids on IL-6 signal transduction in Sko 007 cells were analyzed by the same methods. Results: IL-6 signal transduction could be antagonized by these anti-sense nucleic acids at different extent. Conclusion: TFs or protein kinases in IL-6 signal transduction pathways can be taken as the target for antagonists design.
To investigate which step of IL-6 signal transduction pathways in myeloma cells can be taken as the acting target of the antagonists for IL-6. Methods: EMSA and immunoprecipitation were used to detect the activation of transcription factors(TFs)-STAT3，NF-IL-6 and protein kinase ERK in a myeloma cell line-Sko-007 by IL-6, then anti-sense expression plasmids and anti-sense ODN for these signal moleculars were constructed and designed,the effects of these anti-sense nucleic acids on IL-6 signal transduction in Sko 007 cells were analyzed by the same methods. Results: IL-6 signal transduction could be antagonized by these anti-sense nucleic acids at different extent. Conclusion: TFs or protein kinases in IL-6 signal transduction pathways can be taken as the target for antagonists design.
2000, 7(2):86-89.
DOI: 10.3872/j.issn.1007-385X.2000.2.003
Abstract:
To determine the effect of Ad-53 and Ad-16both alone and in combination on the growth of human lung adeuocarcinoma cell line GLC-82. Methods: Human lung adenocarcinoma cell line GLC-82 were trasferredvia an adenovirus-mediated p53 gene (Ad-p53), P16 gene Ad-p16 or both. Cell growth inhibition assay, colony formation, flow cytometry assay, TUNEL, RT-PCR and immunhistochemistry assays were used to evaluate the therapeutic efficiency of such strategy on trasferred cells. Results: in vitro experiments, at the same dose ofAd (multiplicity of infection), simultaneous transfer of Ad-p53 and Ad-p16 to GLC-82 cells caused stronger therapeutic efficacy than caused by single kind of Ad transfer, demonstrating that Ad-p53 treatment has synergistic efficacy with Ad-p16 treatment on GLC-82 cells. Conclusion: Ad-p53 for GLC-82 cells shows enhanced therapeutic efficiency when combined with Ad-p16.
To determine the effect of Ad-53 and Ad-16both alone and in combination on the growth of human lung adeuocarcinoma cell line GLC-82. Methods: Human lung adenocarcinoma cell line GLC-82 were trasferredvia an adenovirus-mediated p53 gene (Ad-p53), P16 gene Ad-p16 or both. Cell growth inhibition assay, colony formation, flow cytometry assay, TUNEL, RT-PCR and immunhistochemistry assays were used to evaluate the therapeutic efficiency of such strategy on trasferred cells. Results: in vitro experiments, at the same dose ofAd (multiplicity of infection), simultaneous transfer of Ad-p53 and Ad-p16 to GLC-82 cells caused stronger therapeutic efficacy than caused by single kind of Ad transfer, demonstrating that Ad-p53 treatment has synergistic efficacy with Ad-p16 treatment on GLC-82 cells. Conclusion: Ad-p53 for GLC-82 cells shows enhanced therapeutic efficiency when combined with Ad-p16.
2000, 7(2):94-97.
DOI: 10.3872/j.issn.1007-385X.2000.2.005
Abstract:
In order to explore the reversal of the multidrug resistance(MDR) purely related to P-glycoprotein expression by using the anti-MDR1 ribozyme genes. Methods： Daudi human lymphoma cells 20-fold resistant to vincristine (VCR)(Daudi/MDR20) were developed. Three retroviral vectors highly expressing the anti-MDR1 ribozymes: 196MDR1-Rz, 196MDR1-sRz and iMDR1-sRz were also constructed and transfected into GP+envAM12 cells for packaging. The viral supernatants with high titers ［(1.1～2.5)×10 5 cfu/ml］ were used to infected Daudi/MDR20 cells. Results： The tRNAimet-iMDR1-sRz and tRNAimet-196MDR1-sRz-transduced Daudi/MDR20 cells effectively restored chemosensitivity to VCR and were accompanied by blocked expression of MDR1 mRNA and P-glycoprotein as well as overexpression of anti-MDR1-Rzs. Conclusion: The high level expressions of anti-MDR1 ribozymes mediated by retroviral vectors appear to have contributed to the efficinet reversal of P-gp-mediated MDR in tumor cells.
In order to explore the reversal of the multidrug resistance(MDR) purely related to P-glycoprotein expression by using the anti-MDR1 ribozyme genes. Methods： Daudi human lymphoma cells 20-fold resistant to vincristine (VCR)(Daudi/MDR20) were developed. Three retroviral vectors highly expressing the anti-MDR1 ribozymes: 196MDR1-Rz, 196MDR1-sRz and iMDR1-sRz were also constructed and transfected into GP+envAM12 cells for packaging. The viral supernatants with high titers ［(1.1～2.5)×10 5 cfu/ml］ were used to infected Daudi/MDR20 cells. Results： The tRNAimet-iMDR1-sRz and tRNAimet-196MDR1-sRz-transduced Daudi/MDR20 cells effectively restored chemosensitivity to VCR and were accompanied by blocked expression of MDR1 mRNA and P-glycoprotein as well as overexpression of anti-MDR1-Rzs. Conclusion: The high level expressions of anti-MDR1 ribozymes mediated by retroviral vectors appear to have contributed to the efficinet reversal of P-gp-mediated MDR in tumor cells.
2000, 7(2):98-101.
DOI: 10.3872/j.issn.1007-385X.2000.2.006
Abstract:
This study was aimed to explore antitumorresponses induced by recombinant vaccinia viruses expressing a point mutant p53 (rVV-p53FL) and enhancive effects of recombinant vaccinia viruses expressing costimulatory molecule B7 (rVV-B7). Methods: A 135 Cys to Tyr point mutant p53 protein was used as the model of tumor associated antigen. rVV-p53FL and rVV-B7 were used as vaccines to test their induction of CTLs and antitumor immunity. Results: Immunization BABL/c mice with rVV-p53FL could elicited specific CD8 + CTLs that could effecively lyse P815-mp53 cells, a transfectant of the murine P815 mastocytoma containing the mutant p53 gene. Treatment with rVV-p53FL could survive a part of mice challenged with 1×10 6 P815-mp53. Treatment with rVV-p53FL could significantly prolong survival of tumor-bearing mice. Admixture at 1∶1 ratio of rVV-p53FL and rVV-B7 could enhance therapeutic antitumor effects of rVV-p53FL. Conclusion: Mutant P53 over-expressed in tumor cells can render cells targets for specific CTLs generated by immunization with mutant P53 protein based vaccine. Costimulatory molecule B7 can enhance tumor-associated antigen inducing antitumor responses.
This study was aimed to explore antitumorresponses induced by recombinant vaccinia viruses expressing a point mutant p53 (rVV-p53FL) and enhancive effects of recombinant vaccinia viruses expressing costimulatory molecule B7 (rVV-B7). Methods: A 135 Cys to Tyr point mutant p53 protein was used as the model of tumor associated antigen. rVV-p53FL and rVV-B7 were used as vaccines to test their induction of CTLs and antitumor immunity. Results: Immunization BABL/c mice with rVV-p53FL could elicited specific CD8 + CTLs that could effecively lyse P815-mp53 cells, a transfectant of the murine P815 mastocytoma containing the mutant p53 gene. Treatment with rVV-p53FL could survive a part of mice challenged with 1×10 6 P815-mp53. Treatment with rVV-p53FL could significantly prolong survival of tumor-bearing mice. Admixture at 1∶1 ratio of rVV-p53FL and rVV-B7 could enhance therapeutic antitumor effects of rVV-p53FL. Conclusion: Mutant P53 over-expressed in tumor cells can render cells targets for specific CTLs generated by immunization with mutant P53 protein based vaccine. Costimulatory molecule B7 can enhance tumor-associated antigen inducing antitumor responses.
2000, 7(2):102-105.
DOI: 10.3872/j.issn.1007-385X.2000.2.007
Abstract:
To set up HSP90β Protein down-regulated digestive tumor cell lines and study the influence of HSP90β to tumor cells. ethods: pcDNA-HSP90 of HSP90β anti-sense RNA reconstructs was transfected into SGC7901 of human gastric cancer cell line, SGC7901/VCR of MDR-type human gastric cancer cell line, HCC7402 of human hepatic cancer cell line and Ec109 of human esophageal cancer cell line by lipofectamine. Positive clones were selected by G418. Antisense RNA of HSP90β of the positive clones was detected by RNase protection assay and the expression of HSP90β protein was assayed by Western blot. Cell growth was studied by cell growth curve. Results: pcDNA-HSP90 transfected cells were named AH-SGC7901, AH-SGC7901/VCR, AH-HCC7402 and AH-Ec109 respectively and there were antisense RNA of HSP90β in them. HSP90β protein in them was lower than in their parental cells. Cell growth was inhibited to different degrees, and the growth of AH-SGC7901/VCR and AH-Ec109 cells were most greatly inhibited. Conclusion: The expression of HSP90β protein in AH-SGC7901, AH-SGC7901/VCR, AH-HCC7402 and AH-Ec109 cell lines was decreased and the study also showed that decreasing the expression of HSP90β could inhibit tumor cell growth.
To set up HSP90β Protein down-regulated digestive tumor cell lines and study the influence of HSP90β to tumor cells. ethods: pcDNA-HSP90 of HSP90β anti-sense RNA reconstructs was transfected into SGC7901 of human gastric cancer cell line, SGC7901/VCR of MDR-type human gastric cancer cell line, HCC7402 of human hepatic cancer cell line and Ec109 of human esophageal cancer cell line by lipofectamine. Positive clones were selected by G418. Antisense RNA of HSP90β of the positive clones was detected by RNase protection assay and the expression of HSP90β protein was assayed by Western blot. Cell growth was studied by cell growth curve. Results: pcDNA-HSP90 transfected cells were named AH-SGC7901, AH-SGC7901/VCR, AH-HCC7402 and AH-Ec109 respectively and there were antisense RNA of HSP90β in them. HSP90β protein in them was lower than in their parental cells. Cell growth was inhibited to different degrees, and the growth of AH-SGC7901/VCR and AH-Ec109 cells were most greatly inhibited. Conclusion: The expression of HSP90β protein in AH-SGC7901, AH-SGC7901/VCR, AH-HCC7402 and AH-Ec109 cell lines was decreased and the study also showed that decreasing the expression of HSP90β could inhibit tumor cell growth.
2000, 7(2):106-108.
DOI: 10.3872/j.issn.1007-385X.2000.2.008
Abstract:
To investigate whether bcl-2 directly contributes to the development of drug resistance and apoptosis in ovarian cancer cell lines cells OC3. Methods: Retrovirus expression vector pLXSN-bcl-2 was constructed and was transfected to ovarian cancer cell line cells OC3 using Lipofectin, with empty vector pLXSN as a control. Bcl-2 expression of transfected cells was analyzed by FACS and Western blot and Adr-induced cytotoxicity was measured by MTT assay. Apoptosis was also detected by PI DNA staining and DNA Ladder analysis.]Results: pLXSN/bcl-2 transfected cells OC3/bcl-2 overexpressed bcl-2 and were resistant to Adr-induced cytotoxicity. The ability against Adr-inducedapoptosis was increased. Conclusion: bcl-2 may play an important role in the resistance to Adr-inducing apoptosis, thereby increasing resistance of OC3/bcl-2 cells to chemotherapy.
To investigate whether bcl-2 directly contributes to the development of drug resistance and apoptosis in ovarian cancer cell lines cells OC3. Methods: Retrovirus expression vector pLXSN-bcl-2 was constructed and was transfected to ovarian cancer cell line cells OC3 using Lipofectin, with empty vector pLXSN as a control. Bcl-2 expression of transfected cells was analyzed by FACS and Western blot and Adr-induced cytotoxicity was measured by MTT assay. Apoptosis was also detected by PI DNA staining and DNA Ladder analysis.]Results: pLXSN/bcl-2 transfected cells OC3/bcl-2 overexpressed bcl-2 and were resistant to Adr-induced cytotoxicity. The ability against Adr-inducedapoptosis was increased. Conclusion: bcl-2 may play an important role in the resistance to Adr-inducing apoptosis, thereby increasing resistance of OC3/bcl-2 cells to chemotherapy.
2000, 7(2):109-111.
DOI: 10.3872/j.issn.1007-385X.2000.2.009
Abstract:
To explore the role of human hematopoietic growth factors (HGFs) in proliferation and differentiation of cord blood CD34 +cells. Methods: Human cord blood mononuclear cells (MNC) were cultured with different combinations of HGFs (including rhIL-1, rhIL-3, rhIL-6, rhG-CSF, rhGM-CSF and rhSCF). The expansion folds of MNC, the changes of cellular surface markers by FACS, and CFU-GM of hematopoietic cells were observed. Results: The total nucleated cells expanded by 44 folds after cultured with 6 cytokines for 20 days. The number of CFU-GM increased by 14.74 folds after cultured in liquid for 8 days, but decreased heavily after 18 days. The total number of CD34+ cells increased by 127.79～196.40 folds at 6 to 8 day, but decreased to 101.51 folds at 18 day. Conclusion: Human hematopoietic growth factors can increase the expansion of cord blood CD34+ cells and CFU-GM significantly.
To explore the role of human hematopoietic growth factors (HGFs) in proliferation and differentiation of cord blood CD34 +cells. Methods: Human cord blood mononuclear cells (MNC) were cultured with different combinations of HGFs (including rhIL-1, rhIL-3, rhIL-6, rhG-CSF, rhGM-CSF and rhSCF). The expansion folds of MNC, the changes of cellular surface markers by FACS, and CFU-GM of hematopoietic cells were observed. Results: The total nucleated cells expanded by 44 folds after cultured with 6 cytokines for 20 days. The number of CFU-GM increased by 14.74 folds after cultured in liquid for 8 days, but decreased heavily after 18 days. The total number of CD34+ cells increased by 127.79～196.40 folds at 6 to 8 day, but decreased to 101.51 folds at 18 day. Conclusion: Human hematopoietic growth factors can increase the expansion of cord blood CD34+ cells and CFU-GM significantly.
2000, 7(2):112-114.
DOI: 10.3872/j.issn.1007-385X.2000.2.010
Abstract:
To evaluate the therapeutic efficacy of retroviral-mediated HSV-tk/GCV system combined with radiotherapy on murine melanoma. Methods: By using retroviral-mediated gene transfer technique， hygromycin phospotransferase-thymidine kinase fusion gene(hytk) was transferred into B16 murine melanoma cells. Antitumor effects were observed after γ-rays irradiation with 60 Co rays as well as GCV treatment. Growth inhibition of melanoma models in C57BL/6 mice by HSV-tk/GCV systems in combination with radiotherapy was observed. Results: B16 cells transduced with hytk gene and treated with low concentrations of GCV， which showed little cytotoxicity alone， were highly sensitive to radiation. The sensitizer enhancement ratio(SER) was 1.62. This radiopsensitization was also obtained in vivo. Conclusion： The addition of retroviral-mediated HSV-tk/GCV gene therapy to standard radiation therapy an improve the therapeutic effectiveness for melanoma.
To evaluate the therapeutic efficacy of retroviral-mediated HSV-tk/GCV system combined with radiotherapy on murine melanoma. Methods: By using retroviral-mediated gene transfer technique， hygromycin phospotransferase-thymidine kinase fusion gene(hytk) was transferred into B16 murine melanoma cells. Antitumor effects were observed after γ-rays irradiation with 60 Co rays as well as GCV treatment. Growth inhibition of melanoma models in C57BL/6 mice by HSV-tk/GCV systems in combination with radiotherapy was observed. Results: B16 cells transduced with hytk gene and treated with low concentrations of GCV， which showed little cytotoxicity alone， were highly sensitive to radiation. The sensitizer enhancement ratio(SER) was 1.62. This radiopsensitization was also obtained in vivo. Conclusion： The addition of retroviral-mediated HSV-tk/GCV gene therapy to standard radiation therapy an improve the therapeutic effectiveness for melanoma.
2000, 7(2):115-117.
DOI: 10.3872/j.issn.1007-385X.2000.2.011
Abstract:
Endostatin is an effective endogenous inhibitor of angiogensis. To lay foundation of studying the further function of endostatin, the endostatin cDNA from Chinese fetus hepatocytes was isolated, cloned and sequenced. Methods: The total mRNA was isolated from Chinese fetus hepatocytes. The endostatin cDNA was synthesized and amplified from the mRNA by RT-PCR and the PCR product was cloned into pGEM-T vector and subjected to DNA sequencing. Results: The endostatin cDNA was successfully cloned and sequenced. Conclusion: The coding sequence of Chinese Endostatin cDNA is the same as reported sequence derived from foreigner.
Endostatin is an effective endogenous inhibitor of angiogensis. To lay foundation of studying the further function of endostatin, the endostatin cDNA from Chinese fetus hepatocytes was isolated, cloned and sequenced. Methods: The total mRNA was isolated from Chinese fetus hepatocytes. The endostatin cDNA was synthesized and amplified from the mRNA by RT-PCR and the PCR product was cloned into pGEM-T vector and subjected to DNA sequencing. Results: The endostatin cDNA was successfully cloned and sequenced. Conclusion: The coding sequence of Chinese Endostatin cDNA is the same as reported sequence derived from foreigner.
2000, 7(2):118-120.
DOI: 10.3872/j.issn.1007-385X.2000.2.012
Abstract:
To investigate the safety of transgenic human lung adenocarcinoma cell line SPC-A-1/IL-2 as tumor vaccine. Methods: IL-2 gene was introduced into human lung adenocarcinoma cell line SPC-A-1 and was expressed stably on the basis of the construction of retroviral packing cell line PA317/pLIL-2SN. The mutagenesis of both the DNA and supernatant of SPC-A-1/IL-2 in vivo and in vitro was tested by means of genetic toxicological techniques. Results: The result indicated that mutagenesis of both the DNA and the supernatant of transgenic cell SPC-A-1/IL-2 was not observed. Conclusion: The initial experiment suggested that the application of transgenic cell SPC-A-1/IL-2 as tumor vaccine was bisically safe and reliable.
To investigate the safety of transgenic human lung adenocarcinoma cell line SPC-A-1/IL-2 as tumor vaccine. Methods: IL-2 gene was introduced into human lung adenocarcinoma cell line SPC-A-1 and was expressed stably on the basis of the construction of retroviral packing cell line PA317/pLIL-2SN. The mutagenesis of both the DNA and supernatant of SPC-A-1/IL-2 in vivo and in vitro was tested by means of genetic toxicological techniques. Results: The result indicated that mutagenesis of both the DNA and the supernatant of transgenic cell SPC-A-1/IL-2 was not observed. Conclusion: The initial experiment suggested that the application of transgenic cell SPC-A-1/IL-2 as tumor vaccine was bisically safe and reliable.
2000, 7(2):121-123.
DOI: 10.3872/j.issn.1007-385X.2000.2.013
Abstract:
To investigate the anti-tumor efficiency of B16 melanoma HACs in vivo and in vitro. Methods: Tris-HCl extract and Sephacryl S-200 gel filtration were applied to prepare B16 HACs, and the cytoloxicity of specific CTL induced by HACs was tested. Results: The 41, 47 and 53 tube HACs obtained by gel filtration could decrease the tumor incidences, delay the time of tumor development and decrease the mortalitites of mice. Conclusion: 60～97 kD HACs from B16 melanoma cytosol have the activites of inhibiting tumor and could be used in effective anti-tumor therapy.
To investigate the anti-tumor efficiency of B16 melanoma HACs in vivo and in vitro. Methods: Tris-HCl extract and Sephacryl S-200 gel filtration were applied to prepare B16 HACs, and the cytoloxicity of specific CTL induced by HACs was tested. Results: The 41, 47 and 53 tube HACs obtained by gel filtration could decrease the tumor incidences, delay the time of tumor development and decrease the mortalitites of mice. Conclusion: 60～97 kD HACs from B16 melanoma cytosol have the activites of inhibiting tumor and could be used in effective anti-tumor therapy.
2000, 7(2):124-126.
DOI: 10.3872/j.issn.1007-385X.2000.2.014
Abstract:
To investigate the effects of adenovirus type 5 E1A gene (Ad5E1A) on chemosensitivity of human lung adenocarcinoma cells. Methods: The recombinant pcDNA3-E1A, a mammalian-expressing vector was introduced into the Anip973 cells with lipofectAMINE. The cells resistant G418 were selected. Dishes (24-well) were inoculated with Anip973, Anip973-vect and Anip973-E1A cells. Subconfluent monolayers were treated with cisplatin, paclitaxel and VP16 for 24 h, respectively. Viability of the cells was quantitated with MTT method. Results: The sensitivity of Anip973-E1A cells to cytotoxic agents (cisplatin and paclitaxel) was dramatically increased. The IC 50 values for cisplatin and paclitaxel were reduced to approximate 7-fold in Anip973-E1A cells as compared with Anip973-vect cells. For VP16, the sensitivity was not changed. Conclusion: The results demonstrated that E1A gene was capable of enhancing the sensitivity of human lung adenocarcinoma cells to cytotoxic agents (cisplatin and paclitaxel), and the increased sensitivity was associated with the reduction of p185 level, suggesting the possibility of using E1A gene combined with anticancer agents to improve tumor treatment.
To investigate the effects of adenovirus type 5 E1A gene (Ad5E1A) on chemosensitivity of human lung adenocarcinoma cells. Methods: The recombinant pcDNA3-E1A, a mammalian-expressing vector was introduced into the Anip973 cells with lipofectAMINE. The cells resistant G418 were selected. Dishes (24-well) were inoculated with Anip973, Anip973-vect and Anip973-E1A cells. Subconfluent monolayers were treated with cisplatin, paclitaxel and VP16 for 24 h, respectively. Viability of the cells was quantitated with MTT method. Results: The sensitivity of Anip973-E1A cells to cytotoxic agents (cisplatin and paclitaxel) was dramatically increased. The IC 50 values for cisplatin and paclitaxel were reduced to approximate 7-fold in Anip973-E1A cells as compared with Anip973-vect cells. For VP16, the sensitivity was not changed. Conclusion: The results demonstrated that E1A gene was capable of enhancing the sensitivity of human lung adenocarcinoma cells to cytotoxic agents (cisplatin and paclitaxel), and the increased sensitivity was associated with the reduction of p185 level, suggesting the possibility of using E1A gene combined with anticancer agents to improve tumor treatment.
2000, 7(2):127-130.
DOI: 10.3872/j.issn.1007-385X.2000.2.015
Abstract:
To investigate the influence of Icarrin(ICA) on transforming growth factor β ２ (TGFβ ２) expression of PG cells and the mechanisms of reversion of TGFβ ２ mediated immunosuppression by ICA. Methods: Cell proliferation and cytotoxicity was assayed by MTT assay. mRNA level of TGFβ ２ and perforin was assayed by RT-PCR assay. Expression of TGFβ ２ and IL-2R α was analysed by folw cytometry assay. Results: ICA could inhibit the protein and mRNA expression of TGFβ ２ in PG cells. It could also reverse the erpressed cytotoxicity of LAK and CD３ＡＫ cells by TGFβ ２. The IL-2R α expression on LAK and perforin gene transcription and proliferation of CD3AK inhibited by TGFβ ２ were also pastly recovered. Conclusion: The anti-tumor activity of ICA is at least partly due to its capability of inhibiting TGFβ 2 expression in tumor cells and restoring the cytotoxicity of immunocompetent cells inhibited by TGFβ ２
To investigate the influence of Icarrin(ICA) on transforming growth factor β ２ (TGFβ ２) expression of PG cells and the mechanisms of reversion of TGFβ ２ mediated immunosuppression by ICA. Methods: Cell proliferation and cytotoxicity was assayed by MTT assay. mRNA level of TGFβ ２ and perforin was assayed by RT-PCR assay. Expression of TGFβ ２ and IL-2R α was analysed by folw cytometry assay. Results: ICA could inhibit the protein and mRNA expression of TGFβ ２ in PG cells. It could also reverse the erpressed cytotoxicity of LAK and CD３ＡＫ cells by TGFβ ２. The IL-2R α expression on LAK and perforin gene transcription and proliferation of CD3AK inhibited by TGFβ ２ were also pastly recovered. Conclusion: The anti-tumor activity of ICA is at least partly due to its capability of inhibiting TGFβ 2 expression in tumor cells and restoring the cytotoxicity of immunocompetent cells inhibited by TGFβ ２
2000, 7(2):131-134.
DOI: 10.3872/j.issn.1007-385X.2000.2.016
Abstract:
To strengthen the immunogenicity of hepatocarcinoma cells and activate immnological cells recognizing and killing the tumor cells. Methods： The murine MHC-Ⅰ gene H-2K b which can express immunologial rejection antigens was transfected into human hepatocarcinoma cells HepG 2 by liposome DNA mediated gene transfer. The transfection and transcription of H-2K b gene were detected by molecular hybridization techniques. The exogeous antigens expressed on the membrane of transfected tumor cells were detected with ABC immunohistochemical method and flow cytometer. ［ 3H］ release assays were used to detect the recognizing and killing effects of lymphocytes to HepG 2 cells transferred with murine H-2K b gene. The nude mice experiment was used to further verify CTL cells killing active. Results: Southern blot hybridization showed that the H-2K b gene was integrated into the chromosome of HepG 2 cells. The RNA dot blot hybridization showed that there was transcription of H-2K b DNA in the transfected tumor cells. ABC immunohistochemical method and flow cytometer detection showed that the murine H-2K b antigens were expressed on the membrane of HepG 2 cells. ［3H］ release assays showed that the cytotoxicyty to HepG 2 cells transfected with H-2Kb gene was obviously higher than that to control cells. The results demonstrated that the growth of hepatocarcinoma cells which were transferred with H-2Kb gene was obviously inhibited. Conclusion： The murine MHC-Ⅰ gene H-2K b could be transferred into the human hepatocarcinoma cells and expressed on the membrane of transferred cells. The HepG 2 cells transferred with H-2K b gene could induce human effective lymphocytes to recognize and kill these transferred tumor cells.
To strengthen the immunogenicity of hepatocarcinoma cells and activate immnological cells recognizing and killing the tumor cells. Methods： The murine MHC-Ⅰ gene H-2K b which can express immunologial rejection antigens was transfected into human hepatocarcinoma cells HepG 2 by liposome DNA mediated gene transfer. The transfection and transcription of H-2K b gene were detected by molecular hybridization techniques. The exogeous antigens expressed on the membrane of transfected tumor cells were detected with ABC immunohistochemical method and flow cytometer. ［ 3H］ release assays were used to detect the recognizing and killing effects of lymphocytes to HepG 2 cells transferred with murine H-2K b gene. The nude mice experiment was used to further verify CTL cells killing active. Results: Southern blot hybridization showed that the H-2K b gene was integrated into the chromosome of HepG 2 cells. The RNA dot blot hybridization showed that there was transcription of H-2K b DNA in the transfected tumor cells. ABC immunohistochemical method and flow cytometer detection showed that the murine H-2K b antigens were expressed on the membrane of HepG 2 cells. ［3H］ release assays showed that the cytotoxicyty to HepG 2 cells transfected with H-2Kb gene was obviously higher than that to control cells. The results demonstrated that the growth of hepatocarcinoma cells which were transferred with H-2Kb gene was obviously inhibited. Conclusion： The murine MHC-Ⅰ gene H-2K b could be transferred into the human hepatocarcinoma cells and expressed on the membrane of transferred cells. The HepG 2 cells transferred with H-2K b gene could induce human effective lymphocytes to recognize and kill these transferred tumor cells.
2000, 7(2):135-138.
DOI: 10.3872/j.issn.1007-385X.2000.2.017
Abstract:
To prepare an immunoconjugate of anti-EGFR monoclonal antibody (EQ75) and adriamycin (ADR) and to investigute it's inhibitory effect on human epidermoid carcinoma (A431) in vitro and in vivo and it's side effect on C57BL/6J mice. Methods: The immunoconjugate (EQ75-ADR) was prepared with animproved glutaraldehyde conjugate method and the cytotoxicity effects of EQ75-ADR on A431 cells and Wish cells were measured by MTT assay. A431 tumor-bearing nude mice and C57BL/6J experimental model were established and were used for testing the inhibitory effects of EQ75-ADR. The side effect of EQ75-ADR was compared with ADR. Results: The EQ75-ADR showed specific cytotoxicity on A431cells, the cytotoxicity of EQ75-ADR was 50-fold, 14-fold and 10-fold more efficient than that of EQ75, ADR and mixture of EQ75 and ADR (EQ75+ADR) respectively. The effect of EQ75-ADR on with cells was 10-fold less efficient than that of ADR and EQ75+ADR. As compared with the cytoxicity effect of EQ75-ADR on Wish cells with A431 cells, it showed 300-fold less efficient. Similar cytotoxicity results were shown in 2 h treatment experimental group. The EQ75-ADR exhibited significant anti-tumor activity on tumor-bearing nude mice, and the inhibition rate is 92.2% (P<0.01). As compared with ADR, it's side effect was much lower. Conclusion: The EQ75-ADR exhibited targeted delivery and striking anti-tumor activity in vitro and in vivo and this approach appears a promising future for clinic purpose.
To prepare an immunoconjugate of anti-EGFR monoclonal antibody (EQ75) and adriamycin (ADR) and to investigute it's inhibitory effect on human epidermoid carcinoma (A431) in vitro and in vivo and it's side effect on C57BL/6J mice. Methods: The immunoconjugate (EQ75-ADR) was prepared with animproved glutaraldehyde conjugate method and the cytotoxicity effects of EQ75-ADR on A431 cells and Wish cells were measured by MTT assay. A431 tumor-bearing nude mice and C57BL/6J experimental model were established and were used for testing the inhibitory effects of EQ75-ADR. The side effect of EQ75-ADR was compared with ADR. Results: The EQ75-ADR showed specific cytotoxicity on A431cells, the cytotoxicity of EQ75-ADR was 50-fold, 14-fold and 10-fold more efficient than that of EQ75, ADR and mixture of EQ75 and ADR (EQ75+ADR) respectively. The effect of EQ75-ADR on with cells was 10-fold less efficient than that of ADR and EQ75+ADR. As compared with the cytoxicity effect of EQ75-ADR on Wish cells with A431 cells, it showed 300-fold less efficient. Similar cytotoxicity results were shown in 2 h treatment experimental group. The EQ75-ADR exhibited significant anti-tumor activity on tumor-bearing nude mice, and the inhibition rate is 92.2% (P<0.01). As compared with ADR, it's side effect was much lower. Conclusion: The EQ75-ADR exhibited targeted delivery and striking anti-tumor activity in vitro and in vivo and this approach appears a promising future for clinic purpose.
2000, 7(2):139-142.
DOI: 10.3872/j.issn.1007-385X.2000.2.018
Abstract:
To evaluate the efficacy and tolerability of rhGMCSF in prevention of neutropenia caused by chemotherapy of cancer patients. Methods: A muticenter, randomized, match and crossover clinical study was conducted. 64 enrolled patients were randomized into AB and BA groups and each patient received two cycles of combination chemotherapy. Zesults: 59 patients were evaluted for clinical efficacy. rhGM-CSF significantly increased the number of WBC and ANC at the stage of nadir. The period when the patients, rhGM-CSF also resulted in a hastening recovery from leucopenia and neutropenia. Conclusion: The administration of rhGM-CSF ensures the implement of sheduled combination chemotherapy. The main side effects such as fever, osteomyalgia, skin rash were generally tolerable.
To evaluate the efficacy and tolerability of rhGMCSF in prevention of neutropenia caused by chemotherapy of cancer patients. Methods: A muticenter, randomized, match and crossover clinical study was conducted. 64 enrolled patients were randomized into AB and BA groups and each patient received two cycles of combination chemotherapy. Zesults: 59 patients were evaluted for clinical efficacy. rhGM-CSF significantly increased the number of WBC and ANC at the stage of nadir. The period when the patients, rhGM-CSF also resulted in a hastening recovery from leucopenia and neutropenia. Conclusion: The administration of rhGM-CSF ensures the implement of sheduled combination chemotherapy. The main side effects such as fever, osteomyalgia, skin rash were generally tolerable.
2000, 7(2):143-145.
DOI: 10.3872/j.issn.1007-385X.2000.2.019
Abstract:
To evaluate the efficacy and side effects of rhG-CSF (recombinant human granulocyte colony stimulating factor) in chemotherapy induced neutropenia. Methods: 63 cases with malignacies confirmed by pathology or cytology were enrolled. All patients were divided into group AB or BA at randomly self-control cross-over test. At the time of 48 hours after chemotherapy, rhG-CSF was given at a dose of 5 μg?kg -1 ?d -1 for 7 to 14 days in group A. In group B, the patients received chemotherapy alone without rhG-CSF as control. Blood routine examination was taken every other day from the start of chemotherapy. The change of absolute neutrophil and neukocyte counts were observed. Results: In the trial group, neutrophil count increased rapidly with the first peak after 48 hours of rhG-CSF injection. The second peak occurred on the tenth day. In the control group, the absolute neutrophil and leukocyte counts decreased gradually, lower than that of the trial group all the time. There is a significant difference between the trial and control groups (P<0.05) on the nadir of WBC and ANC counts, the duration of WBC<4.0×10 9/L and ANC<2.0×10 9/L, and the number of patients with WBC<4.0×10 9/L on the twenty-first day after chemotherapy. The rates of bone pain and local response of injection were 25.8% and 27.4% respectively. Influenzal symptom was infrequent. Conclusion: rhG-CSF plays an important role in accelerating the recovery of leukocyte and absolute neutrophil induced by chemotherapy. The patients were well toleranced. It is suggested that rhG-CSF be a favourably adjuvant drug in the high dose intensive chemotherapy.
To evaluate the efficacy and side effects of rhG-CSF (recombinant human granulocyte colony stimulating factor) in chemotherapy induced neutropenia. Methods: 63 cases with malignacies confirmed by pathology or cytology were enrolled. All patients were divided into group AB or BA at randomly self-control cross-over test. At the time of 48 hours after chemotherapy, rhG-CSF was given at a dose of 5 μg?kg -1 ?d -1 for 7 to 14 days in group A. In group B, the patients received chemotherapy alone without rhG-CSF as control. Blood routine examination was taken every other day from the start of chemotherapy. The change of absolute neutrophil and neukocyte counts were observed. Results: In the trial group, neutrophil count increased rapidly with the first peak after 48 hours of rhG-CSF injection. The second peak occurred on the tenth day. In the control group, the absolute neutrophil and leukocyte counts decreased gradually, lower than that of the trial group all the time. There is a significant difference between the trial and control groups (P<0.05) on the nadir of WBC and ANC counts, the duration of WBC<4.0×10 9/L and ANC<2.0×10 9/L, and the number of patients with WBC<4.0×10 9/L on the twenty-first day after chemotherapy. The rates of bone pain and local response of injection were 25.8% and 27.4% respectively. Influenzal symptom was infrequent. Conclusion: rhG-CSF plays an important role in accelerating the recovery of leukocyte and absolute neutrophil induced by chemotherapy. The patients were well toleranced. It is suggested that rhG-CSF be a favourably adjuvant drug in the high dose intensive chemotherapy.
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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
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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.