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Volume 5,Issue 2,1998 Table of Contents
1998, 5(2):90-92.
DOI: 10.3872/j.issn.1007-385X.1998.2.004
Abstract:
本实验用RT-PCR方法得到小鼠γ-干扰素(mIFN-γ)全长cDNA,通过逆转录病毒载体将其导入小鼠肝癌细胞株,比较研究了该株细胞在mIFN-γ基因修饰前后致瘤性的变化,并制备了瘤苗,对其分泌干扰素的能力及对荷瘤小鼠的治疗作用作了进一步的研究,发现经过mIFN-γ基因修饰后,肿瘤细胞的致瘤性明显降低且瘤苗对荷瘤小鼠有明显的治疗作用,这为进一步开展细胞因子基因治疗工作打下一定基础。
本实验用RT-PCR方法得到小鼠γ-干扰素(mIFN-γ)全长cDNA,通过逆转录病毒载体将其导入小鼠肝癌细胞株,比较研究了该株细胞在mIFN-γ基因修饰前后致瘤性的变化,并制备了瘤苗,对其分泌干扰素的能力及对荷瘤小鼠的治疗作用作了进一步的研究,发现经过mIFN-γ基因修饰后,肿瘤细胞的致瘤性明显降低且瘤苗对荷瘤小鼠有明显的治疗作用,这为进一步开展细胞因子基因治疗工作打下一定基础。
1998, 5(2):93-96.
DOI: 10.3872/j.issn.1007-385X.1998.2.005
Abstract:
In order to demonstrate the immune regulatory function of human prolactin and its potential in adoptive im-munotherapy, CB17 SCID mice were loaded with human colon adenocarcinoma (HT29) i.p. and then injected i.p. with nylon wool purified human T and NK cells from PBL, followed by rhPRL treatment. The results showed that rhPRL did not exert direct inhibitory ffects on HT-29 cell, in contrast to it, improved the tumor cell growth both in vitro and in vivo . After SCID mice reconstituted with human T and NK cells, rhPRL increased the antitumor effects of human T and NK cells in HT-29-bearing SCID mice, demonstrating the prolonged survival period from 70.4 days to 112.1 days, and the increased survival rate from all died to 60% survival at day 105 compared with HBSS group. In vitro experiments, it was found that rhPRL might improve the proliferation of human T and NK cells, the supernatant of which may inhibit the growth of HT-29 cells in vitro, and improve the cytotoxicity of human T and NK cells against HT-29 tumor cells in 4-hour coculture.
In order to demonstrate the immune regulatory function of human prolactin and its potential in adoptive im-munotherapy, CB17 SCID mice were loaded with human colon adenocarcinoma (HT29) i.p. and then injected i.p. with nylon wool purified human T and NK cells from PBL, followed by rhPRL treatment. The results showed that rhPRL did not exert direct inhibitory ffects on HT-29 cell, in contrast to it, improved the tumor cell growth both in vitro and in vivo . After SCID mice reconstituted with human T and NK cells, rhPRL increased the antitumor effects of human T and NK cells in HT-29-bearing SCID mice, demonstrating the prolonged survival period from 70.4 days to 112.1 days, and the increased survival rate from all died to 60% survival at day 105 compared with HBSS group. In vitro experiments, it was found that rhPRL might improve the proliferation of human T and NK cells, the supernatant of which may inhibit the growth of HT-29 cells in vitro, and improve the cytotoxicity of human T and NK cells against HT-29 tumor cells in 4-hour coculture.
1998, 5(2):105-108.
DOI: 10.3872/j.issn.1007-385X.1998.2.008
Abstract:
The plysical and chemical characteristics of recombinant human GM - CSF (rhGM - CSF) were studied separatly. rhGM - CSF was treated by GdHCl, reduced by DTT, and the disulfide bond was blocked by idoacetamide. The results showed that the samples aren' t homogeneous in UV absorption spectrum and RP-HPLC analysis after treatment by DTT. There was no remarkable differences in the results of the analysis of SDS-PAGE electrophoresis and western blot between treated and untreated rhGM - CSF samples. The effect of GdHCl on GM-CSF was reversible in all above tests; In peptide mapping analysis, the digestion of the samples with blocked disulfide bond by CNBr and protease is more complete than that without any treatment.
The plysical and chemical characteristics of recombinant human GM - CSF (rhGM - CSF) were studied separatly. rhGM - CSF was treated by GdHCl, reduced by DTT, and the disulfide bond was blocked by idoacetamide. The results showed that the samples aren' t homogeneous in UV absorption spectrum and RP-HPLC analysis after treatment by DTT. There was no remarkable differences in the results of the analysis of SDS-PAGE electrophoresis and western blot between treated and untreated rhGM - CSF samples. The effect of GdHCl on GM-CSF was reversible in all above tests; In peptide mapping analysis, the digestion of the samples with blocked disulfide bond by CNBr and protease is more complete than that without any treatment.
1998, 5(2):109-111.
DOI: 10.3872/j.issn.1007-385X.1998.2.009
Abstract:
In older to study the interrelationship between laryngeal carcinoma and human papillomavirus (HPV). The distribution and expression of genome types of HPV in laryngeal carcinoma were investigated. HPV DNA of 160 cases of fresh tissue samples with different lesions of the larynx was detected by applying consensus primers and multiple primers of polymerase chain reaction (PCR) . The consensus primers used were able to detect 9 types of HPVs DNAs such as HPV6, 11, 16, 18, 31, 33, 34, 42 and 58. The results turned out to be as follows: The rate of positive cases with HPV infection was 49.3% (35/71) for the group of laryneal carcinoma, 22.7% (5/22) for the group of neck metastatic lymphnode, 11.8% (2/17) for precarcinomous lesions and 6.7% (2/30 ) for the group of vocal cord polypus, 20 case of normal laryngeal tissue adjacent to the carcinoma was HPV DNA negative. The results demonstrated that the occurrence and development of laryngeal carcinoma were closely related to HPV infection.
In older to study the interrelationship between laryngeal carcinoma and human papillomavirus (HPV). The distribution and expression of genome types of HPV in laryngeal carcinoma were investigated. HPV DNA of 160 cases of fresh tissue samples with different lesions of the larynx was detected by applying consensus primers and multiple primers of polymerase chain reaction (PCR) . The consensus primers used were able to detect 9 types of HPVs DNAs such as HPV6, 11, 16, 18, 31, 33, 34, 42 and 58. The results turned out to be as follows: The rate of positive cases with HPV infection was 49.3% (35/71) for the group of laryneal carcinoma, 22.7% (5/22) for the group of neck metastatic lymphnode, 11.8% (2/17) for precarcinomous lesions and 6.7% (2/30 ) for the group of vocal cord polypus, 20 case of normal laryngeal tissue adjacent to the carcinoma was HPV DNA negative. The results demonstrated that the occurrence and development of laryngeal carcinoma were closely related to HPV infection.
1998, 5(2):112-115.
DOI: 10.3872/j.issn.1007-385X.1998.2.010
Abstract:
In order to investigate the effect of anti-hepato-cellular carcinoma of HSV-tk suicide gene system, we constructed the HSV - tk recombinant retroviral vector pLXT. SMMC - 7721 hepatocellular carconoma cells more transfected with pLXT by lipofectin were obtained by subsequent G418 screen. 3H-TdR incorparation assay showed that HSV - tk/ACV had strong cytotoxic effect on HSV - tk gene transfected tumor cells. Lipofectin pLXT complex was directly injected into murine H22 hepatoma tissue, followed by delivery of ACV prodrup, and it was found that the tumor growth masses more greatly reduced. Animals treated with Liptk ACV and tk ACV had an apparent reduction of tumor size as compared with the animals in other six groups ( P < 0.001, P < 0.05). In addition, there was a significant reduction of tumor size between animals treated with Liptk ACV and tk ACV ( P < 0.02). No significant difference was observed in terms of growth rate of tumors among other six control groups. These results indicate the approach that the recombinant retroviral vector was transduced by cationic liposome is simple, safe and efficient, and could make HSV - tk gene acquire persistence of expression in HCC cells. HSV - tk/ACV system showed strong cytotoxic effect, which suggested to has a strong bystander effect in vivo. In addition, it is efficient that nude HSV-tk gene is directly injected into hepatocellular carcinoma tissue.
In order to investigate the effect of anti-hepato-cellular carcinoma of HSV-tk suicide gene system, we constructed the HSV - tk recombinant retroviral vector pLXT. SMMC - 7721 hepatocellular carconoma cells more transfected with pLXT by lipofectin were obtained by subsequent G418 screen. 3H-TdR incorparation assay showed that HSV - tk/ACV had strong cytotoxic effect on HSV - tk gene transfected tumor cells. Lipofectin pLXT complex was directly injected into murine H22 hepatoma tissue, followed by delivery of ACV prodrup, and it was found that the tumor growth masses more greatly reduced. Animals treated with Liptk ACV and tk ACV had an apparent reduction of tumor size as compared with the animals in other six groups ( P < 0.001, P < 0.05). In addition, there was a significant reduction of tumor size between animals treated with Liptk ACV and tk ACV ( P < 0.02). No significant difference was observed in terms of growth rate of tumors among other six control groups. These results indicate the approach that the recombinant retroviral vector was transduced by cationic liposome is simple, safe and efficient, and could make HSV - tk gene acquire persistence of expression in HCC cells. HSV - tk/ACV system showed strong cytotoxic effect, which suggested to has a strong bystander effect in vivo. In addition, it is efficient that nude HSV-tk gene is directly injected into hepatocellular carcinoma tissue.
Li Jing
,
Wang Zhuozhi
,
Wang Yan
,
Liu Qunying
,
Hua Bing
,
Chen Yuping
,
Zhu Yingchun
,
Dong Zhiwei
1998, 5(2):116-120.
DOI: 10.3872/j.issn.1007-385X.1998.2.011
Abstract:
In order to reduce its immunogenicity and increase its therapeutic usefullness, the murine anti-gastric cancer McAb 3H11 was genetically engineered into a human-mouse chimeric antibody. The 3H11 light and heavy chain variable genes were inserted into chimeric antibody expression vectors respectively. The chimeric light chain expression vector was first transfected into murine Sp2/0 myeloma cells using lipofectin. Transfectants that stably expressed human-mouse chimeric light chain were isolated by mycophenolic acid selection. Then the chimeric heavy chain expression vector was transfected into the transfectomas. Histidinol resistent cells were obtained and among these, clones stably secreting human-mouse chimeric antibodies were selected. The expression of human-mouse chimeric light and heavy chain mRNAs were proved by RT-PCR and the expressed chimeric antibody was demonstrated capable of binding to human gastric cancer cell MGC803
In order to reduce its immunogenicity and increase its therapeutic usefullness, the murine anti-gastric cancer McAb 3H11 was genetically engineered into a human-mouse chimeric antibody. The 3H11 light and heavy chain variable genes were inserted into chimeric antibody expression vectors respectively. The chimeric light chain expression vector was first transfected into murine Sp2/0 myeloma cells using lipofectin. Transfectants that stably expressed human-mouse chimeric light chain were isolated by mycophenolic acid selection. Then the chimeric heavy chain expression vector was transfected into the transfectomas. Histidinol resistent cells were obtained and among these, clones stably secreting human-mouse chimeric antibodies were selected. The expression of human-mouse chimeric light and heavy chain mRNAs were proved by RT-PCR and the expressed chimeric antibody was demonstrated capable of binding to human gastric cancer cell MGC803
Ju Dian Wen
,
Cao Xuetao
,
Wang Baomei
,
Kong Lingfei
,
Yin Pingzhang
,
Tao Qun
,
Wan Tao
,
Yu Yizhi
1998, 5(2):121-125.
DOI: 10.3872/j.issn.1007-385X.1998.2.012
Abstract:
Antitumor effect of combined transfer of suicide gene and cytokine gene was evaluated in the present study. Adenoviruses expressing E. coli. cytosine deaminase (AdCD) and adenoviruses expressing murine interleukin 2 (AdTL2) were used for the treatment of tumor-bearing mice. The mice were inoculated s. c. with FBL-3 leukemia cells and 3 days later received intratumoral injection of AdCD in the presence or absence of AdIL2 followed by intraperitoneal 5-fluorocytosine (5FC) administration. The results demonstrated that tumor-bearing mice treated with AdCD/5FC in combination with AdTL2 showed more .potent inhibition of tumor growth and survived much longer as compared with mice treated with AdCD/5FC, AdEL2, AdlacZ/5FC or PBS. It was illustrated that the tumor mass showed obvious necrosis and inflammatory cell infiltration, and more CD4 and CD8 T cells infiltrated into the tumor after combined therapy. The splenic NK and CTL activities increased significantly in mice after combined transfer of CD gene and EH gene. Our results demonstrated that combined transfer of suicide gene and IL-2 gene could inhibit the growth of established tumor in mice significantly and induce antitumor immunity of the host efficiently.
Antitumor effect of combined transfer of suicide gene and cytokine gene was evaluated in the present study. Adenoviruses expressing E. coli. cytosine deaminase (AdCD) and adenoviruses expressing murine interleukin 2 (AdTL2) were used for the treatment of tumor-bearing mice. The mice were inoculated s. c. with FBL-3 leukemia cells and 3 days later received intratumoral injection of AdCD in the presence or absence of AdIL2 followed by intraperitoneal 5-fluorocytosine (5FC) administration. The results demonstrated that tumor-bearing mice treated with AdCD/5FC in combination with AdTL2 showed more .potent inhibition of tumor growth and survived much longer as compared with mice treated with AdCD/5FC, AdEL2, AdlacZ/5FC or PBS. It was illustrated that the tumor mass showed obvious necrosis and inflammatory cell infiltration, and more CD4 and CD8 T cells infiltrated into the tumor after combined therapy. The splenic NK and CTL activities increased significantly in mice after combined transfer of CD gene and EH gene. Our results demonstrated that combined transfer of suicide gene and IL-2 gene could inhibit the growth of established tumor in mice significantly and induce antitumor immunity of the host efficiently.
Wang Baomei
,
Cao Xuetao
,
Ju Dian Wen
,
Yin Pingzhang
,
Kong Lingfei
,
Wan Tao
,
Tao Qun
,
Yu Yizhi
1998, 5(2):126-129.
DOI: 10.3872/j.issn.1007-385X.1998.2.013
Abstract:
Adenoviruses harboring E. coli cytosine deaminase gene (AdCD) were used to transfect murine FBL-3 ery-throleukemia cells in vitro. FBL3 cells infected with AdCD were more sensitive to 5-fluorocytosine (5-FC) than cells infected with a control adenovirus AdLacZ. Further study indicated that this combination therapy (AdCD and 5-FC) killed tumor cells by inducing apoptosis of FBL-3 cells. The supematants from FBL-3 cells treated with AdCD/5-Fc were transferred on the culture system of uninfected (wild - type) FBL-3 cells, the result indicated that only 6.25% of the supernatant could induce significant cytotoxicity on wild type FBL3 cells. The results demonoustrated that bystander effect plays an important role in AdCD-mediated cytotoxicities. Direct injection of AdCD into established subcutaneous FBL3 tumor in mice followed by daily intraperitoneal injection of 5-FC for 10 days was found to inhibit tumor growth significant-
Adenoviruses harboring E. coli cytosine deaminase gene (AdCD) were used to transfect murine FBL-3 ery-throleukemia cells in vitro. FBL3 cells infected with AdCD were more sensitive to 5-fluorocytosine (5-FC) than cells infected with a control adenovirus AdLacZ. Further study indicated that this combination therapy (AdCD and 5-FC) killed tumor cells by inducing apoptosis of FBL-3 cells. The supematants from FBL-3 cells treated with AdCD/5-Fc were transferred on the culture system of uninfected (wild - type) FBL-3 cells, the result indicated that only 6.25% of the supernatant could induce significant cytotoxicity on wild type FBL3 cells. The results demonoustrated that bystander effect plays an important role in AdCD-mediated cytotoxicities. Direct injection of AdCD into established subcutaneous FBL3 tumor in mice followed by daily intraperitoneal injection of 5-FC for 10 days was found to inhibit tumor growth significant-
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Supported by:Beijing E-Tiller Technology Development Co., Ltd.