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Volume 11,Issue 3,2004 Table of Contents
2004, 11(3):157-160.
DOI: 10.3872/j.issn.1007-385X.2004.3.001
Abstract:
Objective：To investigate inhibitory effect of specific cytotoxicity lymphocytes(CTLs) pulsed by EBV-LMP2-peptide epitopes on transplantable tumor of nasopharyngeal carcinoma cell (NPC)line-CNE2 with identical HLA-Ⅰ molecule subtypes, and explore the possibility of immunotherapy among indivduals. Methods：Transplantation tumor was established through subcutaneous injection of CNE2 cells in Scid mice. HLA-Ⅰ subtypes in CNE2 cells were identified by specific site primer polymerase chain reaction (SSP-PCR). Matched HLA HLA-Ⅰmolecule subtypes with CNE2 were screened among normal volunteers. Specific CTLs stimulated by EBV-LMP2-peptide epitopes were collected for the next steps. Three groups were divided. (1) CTL group: CNE2 cells with specific CTLs stimulated by EBV-LMP2-peptide epitopes were injected into mice subcutaneousness. (2) T group: CNE2 cells with t lymphocytes were injected. (3) The control group, CNE2 cells were injected only. The results were observed in a few weeks later. Results：Transplantable tumors were observed after CNE2 having been injected into scids mice. HLA-A11 in CNE2 cells was detected by SSP-PCR. Comparing the results from the three groups, we found specific CTLs stimulated by A11-LMP2-pepetide epitopes distinctly retarded the growth of CNE2 transplantable tumor. Conclusion：Specific-LMP2 CTLs with HLA-A11 molecule subtype effectively inhibit the development of transplantable tumor of CNE2 cells with identical HLA-Ⅰ molecule subtype of three different groups were injected.
Objective：To investigate inhibitory effect of specific cytotoxicity lymphocytes(CTLs) pulsed by EBV-LMP2-peptide epitopes on transplantable tumor of nasopharyngeal carcinoma cell (NPC)line-CNE2 with identical HLA-Ⅰ molecule subtypes, and explore the possibility of immunotherapy among indivduals. Methods：Transplantation tumor was established through subcutaneous injection of CNE2 cells in Scid mice. HLA-Ⅰ subtypes in CNE2 cells were identified by specific site primer polymerase chain reaction (SSP-PCR). Matched HLA HLA-Ⅰmolecule subtypes with CNE2 were screened among normal volunteers. Specific CTLs stimulated by EBV-LMP2-peptide epitopes were collected for the next steps. Three groups were divided. (1) CTL group: CNE2 cells with specific CTLs stimulated by EBV-LMP2-peptide epitopes were injected into mice subcutaneousness. (2) T group: CNE2 cells with t lymphocytes were injected. (3) The control group, CNE2 cells were injected only. The results were observed in a few weeks later. Results：Transplantable tumors were observed after CNE2 having been injected into scids mice. HLA-A11 in CNE2 cells was detected by SSP-PCR. Comparing the results from the three groups, we found specific CTLs stimulated by A11-LMP2-pepetide epitopes distinctly retarded the growth of CNE2 transplantable tumor. Conclusion：Specific-LMP2 CTLs with HLA-A11 molecule subtype effectively inhibit the development of transplantable tumor of CNE2 cells with identical HLA-Ⅰ molecule subtype of three different groups were injected.
2004, 11(3):161-165.
DOI: 10.3872/j.issn.1007-385X.2004.3.002
Abstract:
Objective: To select anti-HAb18G (hepatoma associated antigen) human Fd fragments with guided selection of L chain of chimeric Fab-HAb18.Methods:The human Fd repertories genes were amplified by RT-PCR from PBMC of hepatoma patients, and cloned into the vector pComb3X with chimeric L chain to construct the human-mouse hybrid Fab phage library. HAb18GE, extracellular region of HAb18G, was used as antigen to screen. The positive clones was obtained by ELISA and FCM with pⅢ-fusion Fab antibody. The DNA sequences were analyzed. Results: A human-mouse Fab antibody library were constructed with 2×10 7 PFU. After 6 rounds panning, 7 positive clones were obtained with ELISA and FCM. And sequences of 2 clones with better affinity were same. The CH1 belonged to the IgG2 class as the parent Fd, and the variable region belonged to VH3 family. Conclusions:Through the construction of the HuMFab phage antibody library and chemeric L chain-guided selection, we get the available human Fd fragments for subsequent research.
Objective: To select anti-HAb18G (hepatoma associated antigen) human Fd fragments with guided selection of L chain of chimeric Fab-HAb18.Methods:The human Fd repertories genes were amplified by RT-PCR from PBMC of hepatoma patients, and cloned into the vector pComb3X with chimeric L chain to construct the human-mouse hybrid Fab phage library. HAb18GE, extracellular region of HAb18G, was used as antigen to screen. The positive clones was obtained by ELISA and FCM with pⅢ-fusion Fab antibody. The DNA sequences were analyzed. Results: A human-mouse Fab antibody library were constructed with 2×10 7 PFU. After 6 rounds panning, 7 positive clones were obtained with ELISA and FCM. And sequences of 2 clones with better affinity were same. The CH1 belonged to the IgG2 class as the parent Fd, and the variable region belonged to VH3 family. Conclusions:Through the construction of the HuMFab phage antibody library and chemeric L chain-guided selection, we get the available human Fd fragments for subsequent research.
2004, 11(3):166-169.
DOI: 10.3872/j.issn.1007-385X.2004.3.003
Abstract:
Objective : To explore the preventive and therapeutic effects of Hsp70-antigen peptide complexes (HACs) on pulmonary metastasis of malanoma B16 cells in mice. Methods:Antigen peptide mixtures were prepared from massive tumors and metastatic foci respctively, and bound with Hsp70 in vitro. The HACs were used to mice to prevent the formation of micrometastatic foci after the injection of B16 cells through tail vein, or treat the existent micrometastatic foci in lung The number of metastatic foci was counted and the cytotoxicity of splenocytes to B16 cells was determined. Results: After immunization with HACs, the number of metastatic foci in mice decreased significantly (P<0.01), and the cytotoxicity of splenocytes to B16 cells was signficantly enhanced (P<0.01). The immuniation with HACS can both prevent the formation of metastatic foci and freat the existent metastatic foci. The immunization with petide mixture from massive tumros produced better therapeutic effect (P<0.01), and induced stronger citotoxicity of splenocytes to B16 cells (0.01
Objective : To explore the preventive and therapeutic effects of Hsp70-antigen peptide complexes (HACs) on pulmonary metastasis of malanoma B16 cells in mice. Methods:Antigen peptide mixtures were prepared from massive tumors and metastatic foci respctively, and bound with Hsp70 in vitro. The HACs were used to mice to prevent the formation of micrometastatic foci after the injection of B16 cells through tail vein, or treat the existent micrometastatic foci in lung The number of metastatic foci was counted and the cytotoxicity of splenocytes to B16 cells was determined. Results: After immunization with HACs, the number of metastatic foci in mice decreased significantly (P<0.01), and the cytotoxicity of splenocytes to B16 cells was signficantly enhanced (P<0.01). The immuniation with HACS can both prevent the formation of metastatic foci and freat the existent metastatic foci. The immunization with petide mixture from massive tumros produced better therapeutic effect (P<0.01), and induced stronger citotoxicity of splenocytes to B16 cells (0.01
2004, 11(3):170-174.
DOI: 10.3872/j.issn.1007-385X.2004.3.004
Abstract:
Objective: To study the anti-tumor efficacy induced by antibodized tumor epitope PDTRP gene immunization. Methods:Three copies of tumor associate gene PDTRP from MUC1 tandem repeats were designed and mimicked the conformation of MUC1 by Insight Ⅱ. The γ1neo-PDTRP plasmid was further constructed, in which the PDTRP target gene was inserted into CDR3 of the γ1-neo vector. The specific humoral and cellular immune responses towards to PDTRP were detected after intraspleen immunized Balb/c mice with γ1neo-PDTRP. And the immune protection assay was also done to observe whether the mice immunized with γ1neo-PDTRP could prolong the survival after tumor challenge. Results: The conformation of three copies of PDTRP mimicked the conformation of MUC1 tandem repeats. The expression of γ1neo-PDTRP could be detected after in vitro transfect. The specific antibody against PDTRP epitope could be induced and increase to a higher titer after intraspleen injection with a γ1neo-PDTRP plasmid. And the specific proliferation and cytotoxic function of lymphocyte were also increased. There is a significant survival from mice immunized with γ1neo-PDTRP against the 4T1-PDTRP tumor challenge.Conclusions: Gene immunization with γ1neo-PDTRP could elicit both humoral and cellular tumor specific immune response and had the protective effect.
Objective: To study the anti-tumor efficacy induced by antibodized tumor epitope PDTRP gene immunization. Methods:Three copies of tumor associate gene PDTRP from MUC1 tandem repeats were designed and mimicked the conformation of MUC1 by Insight Ⅱ. The γ1neo-PDTRP plasmid was further constructed, in which the PDTRP target gene was inserted into CDR3 of the γ1-neo vector. The specific humoral and cellular immune responses towards to PDTRP were detected after intraspleen immunized Balb/c mice with γ1neo-PDTRP. And the immune protection assay was also done to observe whether the mice immunized with γ1neo-PDTRP could prolong the survival after tumor challenge. Results: The conformation of three copies of PDTRP mimicked the conformation of MUC1 tandem repeats. The expression of γ1neo-PDTRP could be detected after in vitro transfect. The specific antibody against PDTRP epitope could be induced and increase to a higher titer after intraspleen injection with a γ1neo-PDTRP plasmid. And the specific proliferation and cytotoxic function of lymphocyte were also increased. There is a significant survival from mice immunized with γ1neo-PDTRP against the 4T1-PDTRP tumor challenge.Conclusions: Gene immunization with γ1neo-PDTRP could elicit both humoral and cellular tumor specific immune response and had the protective effect.
2004, 11(3):175-178.
DOI: 10.3872/j.issn.1007-385X.2004.3.005
Abstract:
Objective:The inhibition on the proliferation of colorectal carcinoma cell strain with different proliferative potential by ATRA was investigated in present study, which would benefit for the therapy of ATRA on colorectal carcinoma. Methods: The proliferation of LS174T and CW-2 colorectal carcinoma cell strain inhibited by ATRA was determined using cell observation, FACS and MTT methods.Results: The growth speed of LS174T cell strain was faster than that of CW-2. ATRA played a significant role on inhibiting the proliferation of LS174T and CW-2 cell strain and inducting the cell differetiation in vitro.Conclusions: ATRA could inhibit the growth of LS174T and CW-2 cell strain. ATRA could inhibit the proliferation of colorectal carcinoma cell and induce cell differentiation to some extent, which was correlated with the concentration of ATRA.
Objective:The inhibition on the proliferation of colorectal carcinoma cell strain with different proliferative potential by ATRA was investigated in present study, which would benefit for the therapy of ATRA on colorectal carcinoma. Methods: The proliferation of LS174T and CW-2 colorectal carcinoma cell strain inhibited by ATRA was determined using cell observation, FACS and MTT methods.Results: The growth speed of LS174T cell strain was faster than that of CW-2. ATRA played a significant role on inhibiting the proliferation of LS174T and CW-2 cell strain and inducting the cell differetiation in vitro.Conclusions: ATRA could inhibit the growth of LS174T and CW-2 cell strain. ATRA could inhibit the proliferation of colorectal carcinoma cell and induce cell differentiation to some extent, which was correlated with the concentration of ATRA.
2004, 11(3):179-182.
DOI: 10.3872/j.issn.1007-385X.2004.3.006
Abstract:
Objective: To clone and express the recombinant human Vasostatin120-180 aa domain and to investigate its activity of inhibiting angiogenesis in CAM. Methods:After amplifying gene of human Vasostatin120-180 aa domain, we subcloned it into pQE30 vector and expressed Vasostatin120-180 aa domain by E.coli. We also tested its ability of inhibiting angiogenesis in CAM. Results: The total gene length of human Vasostatin120-180 aa domain is 180 bp. Expressed by pQE30 system in E.coli and purified by IMAC, Vasostatin120-180 aa was detected by SDS-PAGE, in which there is a positive band and molecular weight is about 8 kD.Conclusions:Recombinant human Vasostatin120-180aa could play effective role in anti-angiogenesis in CAM and it showed a dose dependent effect in some degree.
Objective: To clone and express the recombinant human Vasostatin120-180 aa domain and to investigate its activity of inhibiting angiogenesis in CAM. Methods:After amplifying gene of human Vasostatin120-180 aa domain, we subcloned it into pQE30 vector and expressed Vasostatin120-180 aa domain by E.coli. We also tested its ability of inhibiting angiogenesis in CAM. Results: The total gene length of human Vasostatin120-180 aa domain is 180 bp. Expressed by pQE30 system in E.coli and purified by IMAC, Vasostatin120-180 aa was detected by SDS-PAGE, in which there is a positive band and molecular weight is about 8 kD.Conclusions:Recombinant human Vasostatin120-180aa could play effective role in anti-angiogenesis in CAM and it showed a dose dependent effect in some degree.
2004, 11(3):183-186.
DOI: 10.3872/j.issn.1007-385X.2004.3.007
Abstract:
Objective:To investigate the effects of antisense oligoxydeonucleotides of Ki67 gene(Ki67-ASODNs) on the proliferation and apoptosis of human renal carcinoma cell line 786-0 cells. Methods: Human renal carcinoma cell line 786-0 cells were treated with Ki67-ASODNs (10, 40 μmol/L).The Ki67 expression of 786-0 cells was detected by immunohistochemical technique and Western blot respectively. The proliferation of 786-0 cells was studied by cell growth curves and 3H-TdR uptade assay. The apoptosis of 786-0 cells was detected by TUNEL assay. Results: The Ki67 expression rates of 786-0 cells treated by Ki67-ASODNs （29.9±0.4, 24.5±1.2）were lower than that of the control group(P<001). The Ki67 protein quantity rates of 786-0 cells treated by Ki67-ASODNs （82.1±2.2, 66.6±4.2）were reduced markedly(P<0.01). The 3H-thymidine incorporation rates of 786-0 cells treated by Ki67-ASODNs（44.5±4.4, 34.5±3.2） were lower than that of the control group(P<0.01).The apoptosis rates of 786-0 cells treated by Ki67 ASODNs（12.7±0.5, 23.1±2.1）were higher than that of the control group（9.3±2.0； P<0.01). Conclusions: Antisense oligoxydeonucleotides of Ki67 could inhibit the proliferation and induce apoptosis by blocking Ki67 expression of human renal carcinoma 786-0 cells.
Objective:To investigate the effects of antisense oligoxydeonucleotides of Ki67 gene(Ki67-ASODNs) on the proliferation and apoptosis of human renal carcinoma cell line 786-0 cells. Methods: Human renal carcinoma cell line 786-0 cells were treated with Ki67-ASODNs (10, 40 μmol/L).The Ki67 expression of 786-0 cells was detected by immunohistochemical technique and Western blot respectively. The proliferation of 786-0 cells was studied by cell growth curves and 3H-TdR uptade assay. The apoptosis of 786-0 cells was detected by TUNEL assay. Results: The Ki67 expression rates of 786-0 cells treated by Ki67-ASODNs （29.9±0.4, 24.5±1.2）were lower than that of the control group(P<001). The Ki67 protein quantity rates of 786-0 cells treated by Ki67-ASODNs （82.1±2.2, 66.6±4.2）were reduced markedly(P<0.01). The 3H-thymidine incorporation rates of 786-0 cells treated by Ki67-ASODNs（44.5±4.4, 34.5±3.2） were lower than that of the control group(P<0.01).The apoptosis rates of 786-0 cells treated by Ki67 ASODNs（12.7±0.5, 23.1±2.1）were higher than that of the control group（9.3±2.0； P<0.01). Conclusions: Antisense oligoxydeonucleotides of Ki67 could inhibit the proliferation and induce apoptosis by blocking Ki67 expression of human renal carcinoma 786-0 cells.
2004, 11(3):187-190.
DOI: 10.3872/j.issn.1007-385X.2004.3.008
Abstract:
Objective: To investigate the inhibitory effect of survivin antisense oligonucleotide (ASODN) on the expression of survivin gene and proliferation of human hepatocellular carcinoma cell line SMMC-7721.Methods: The 20 mer antisense oligonucleotide (ASODN) targeted to the promotor region of survivin mRNA was designed and synthesized. The expression of survivin gene in hepatocellular carcinoma cell line SMMC-7721 was blocked by means of ASODN transfection mediated by DOTAP liposomal reagent. The changes of survivin protein and mRNA expression after transfection were assessd by Western blot and in situ hybridization, respectively. The apoptotic rate was detected by flow cytometer. The changes of cell adherent rate, cell growth activity, and the inhibitory rate of cell growth were also studied.Results: The expression of survivin protein and mRNA were decreased markedly after survivin ASODN transfection. Meanwhile, the cell adherent rate also decreased markedly while the apoptotic rate increased markedly. Conclusions: Transfection of ASODN targeted to the promotor region of survivin mRNA by DOTAP liposomal transfection reagent could down-regulated the expression of survivin protein and mRNA significantly in 7721 cell line and inhibit the proliferation of cancer cells. Survivin could be an important target in the therapy of hepatocellular carcinoma.
Objective: To investigate the inhibitory effect of survivin antisense oligonucleotide (ASODN) on the expression of survivin gene and proliferation of human hepatocellular carcinoma cell line SMMC-7721.Methods: The 20 mer antisense oligonucleotide (ASODN) targeted to the promotor region of survivin mRNA was designed and synthesized. The expression of survivin gene in hepatocellular carcinoma cell line SMMC-7721 was blocked by means of ASODN transfection mediated by DOTAP liposomal reagent. The changes of survivin protein and mRNA expression after transfection were assessd by Western blot and in situ hybridization, respectively. The apoptotic rate was detected by flow cytometer. The changes of cell adherent rate, cell growth activity, and the inhibitory rate of cell growth were also studied.Results: The expression of survivin protein and mRNA were decreased markedly after survivin ASODN transfection. Meanwhile, the cell adherent rate also decreased markedly while the apoptotic rate increased markedly. Conclusions: Transfection of ASODN targeted to the promotor region of survivin mRNA by DOTAP liposomal transfection reagent could down-regulated the expression of survivin protein and mRNA significantly in 7721 cell line and inhibit the proliferation of cancer cells. Survivin could be an important target in the therapy of hepatocellular carcinoma.
2004, 11(3):191-194.
DOI: 10.3872/j.issn.1007-385X.2004.3.009
Abstract:
Objective:To study the promoting effect of vascular endothelial growth factor (VEGF) on the generation of CD34+ cells from mouse embryonic stem cell(ESC) in vitro.Methods: ES-D3 cells were allowed to grow on mouse fetal fibroblast feeder layer to form embryoid bodies(EB), then EB cells were transferred into medium supplemented with different concentration of VEGF and VEGF+SCF for 1 week. Six groups were designed , i.e. VEGF 5 μg/L, VEGF 10 μg/L, VEGF 20 μg/L, VEGF 5 μg/L＋SCF, VEGF 10 μg/L＋SCF and VEGF 20 μg/L＋SCF. The group of spontaneous differentiation without cytokine(s) was served as control. The expression of CD34 mRNA was detected by RT-PCR. The content of CD34+ cells in each group was measured by flow cytometry. The cells derived from ESC were incubated in semisolid methycellulose cultures. The numbers of total colony-forming units in culture(CFU-C) were counted by reverse microscope. Results：ES-D3 grew and formed EBs at day 4. VEGF gave the stimulatory effects on the expression of CD34 mRNA, the generation of CD34+ cells and CFU-C in EB as a single factor . The effects of VEGF＋SCF were the most effective in the experimental groups according to the percent of CD34+ cells and the numbers of hematopoietic colonies. The most highest inducing efficacy were achieved when VEGF 20 μg/L or VEGF 10 μg/L combined with SCF were used.Conclusion: VEGF can promote the differentiation of ESC into CD34+ cells in vitro. The strongest effects were achieved when VEGF was used in combination with SCF.
Objective:To study the promoting effect of vascular endothelial growth factor (VEGF) on the generation of CD34+ cells from mouse embryonic stem cell(ESC) in vitro.Methods: ES-D3 cells were allowed to grow on mouse fetal fibroblast feeder layer to form embryoid bodies(EB), then EB cells were transferred into medium supplemented with different concentration of VEGF and VEGF+SCF for 1 week. Six groups were designed , i.e. VEGF 5 μg/L, VEGF 10 μg/L, VEGF 20 μg/L, VEGF 5 μg/L＋SCF, VEGF 10 μg/L＋SCF and VEGF 20 μg/L＋SCF. The group of spontaneous differentiation without cytokine(s) was served as control. The expression of CD34 mRNA was detected by RT-PCR. The content of CD34+ cells in each group was measured by flow cytometry. The cells derived from ESC were incubated in semisolid methycellulose cultures. The numbers of total colony-forming units in culture(CFU-C) were counted by reverse microscope. Results：ES-D3 grew and formed EBs at day 4. VEGF gave the stimulatory effects on the expression of CD34 mRNA, the generation of CD34+ cells and CFU-C in EB as a single factor . The effects of VEGF＋SCF were the most effective in the experimental groups according to the percent of CD34+ cells and the numbers of hematopoietic colonies. The most highest inducing efficacy were achieved when VEGF 20 μg/L or VEGF 10 μg/L combined with SCF were used.Conclusion: VEGF can promote the differentiation of ESC into CD34+ cells in vitro. The strongest effects were achieved when VEGF was used in combination with SCF.
CHEN Shao-hua
,
LI Yang-qiu
,
YANG Li-jian
,
HUANG Mei-juan
,
HAN Su-fang
,
LUO Geng-xin
,
CHEN Sheng-ting
2004, 11(3):195-199.
DOI: 10.3872/j.issn.1007-385X.2004.3.010
Abstract:
Objective: To investigate the clonal expansion and specific cytotoxicity of TCR Vβ subfamily T cells from healthy individuals induced by acute myelogenous leukemia M2a subtype (AML-M2a) cells, respectively.Methods: T cells from three healthy individuals that were induced by AML-M2a cells by using mixed lymphocyte and tumor culture (MLTC) in different amplified stage were detected for the specific cytotoxicity by LDH release assay. The distribution and clonal expansion of TCR Vβ 24 subfamily genes of T cells before and after induction were analyzed by RT-PCR and genescan.Results: At 12th and 21st day after culture the cytotoxicity rates of three healthy individuals T cells after MLTC for the induced cells. (AML-M2a cells) were 39.88±7.79% and 62.14±9.79%, respectively, whereas there were not any cytotoxicity in the T cells of peripheral blood from the AML-M2a case and three healthy individuals. Clonal expansion of TCR Vβ16 subfamily T cells was displayed by genescan in the T cells from three donors after MLTC.Conclusions: The clonal expansion and specific cytotoxicity T cells from healthy individuals could be induced by AML-M2a cells. The clonal expansion T cells were more tended to TCR Vβ16 subfamily.
Objective: To investigate the clonal expansion and specific cytotoxicity of TCR Vβ subfamily T cells from healthy individuals induced by acute myelogenous leukemia M2a subtype (AML-M2a) cells, respectively.Methods: T cells from three healthy individuals that were induced by AML-M2a cells by using mixed lymphocyte and tumor culture (MLTC) in different amplified stage were detected for the specific cytotoxicity by LDH release assay. The distribution and clonal expansion of TCR Vβ 24 subfamily genes of T cells before and after induction were analyzed by RT-PCR and genescan.Results: At 12th and 21st day after culture the cytotoxicity rates of three healthy individuals T cells after MLTC for the induced cells. (AML-M2a cells) were 39.88±7.79% and 62.14±9.79%, respectively, whereas there were not any cytotoxicity in the T cells of peripheral blood from the AML-M2a case and three healthy individuals. Clonal expansion of TCR Vβ16 subfamily T cells was displayed by genescan in the T cells from three donors after MLTC.Conclusions: The clonal expansion and specific cytotoxicity T cells from healthy individuals could be induced by AML-M2a cells. The clonal expansion T cells were more tended to TCR Vβ16 subfamily.
11 Anti-Tumor Effect of Repeated Low Dose pEgr-IFN-γ Gene-Radiotherapy and Its Immunologic Mechanism
2004, 11(3):200-203.
DOI: 10.3872/j.issn.1007-385X.2004.3.011
Abstract:
Objective：To study the anti-tumor effect of repeated low dose pEgr-IFN-γ gene-radiotherapy in mice bearing melanoma and its immunologic mechanism. Methods:The pEgr-IFN-γ plasmids packed by liposome were injected locally into tumors, and then the tumors was irradiated by X-ray 36 hours later. The tumor growth rate at different time and mean survival period of the mice were observed. The IFN-γ mRNA level in tumors was detected by RT-PCR 3 days after irradiation. The concentration of IFN-γ in serum were detected by ELISA. The immunologic indexes was detected 15 days after irradiation. Results: The tumor growth rate of the mice in gene-radiotherapy group with pEgr-IFN-γ and 5 Gy X-ray irradiation four times were lower significantly than that with pEgr-IFN-γ and 20 Gy X-ray irradiation 9～15 days after irradiation and their mean survival period were longer. The IFN-γ mRNA level in tumors of the mice in gene-radiotherapy group were higher than that in reconstruction plasmids group significantly 3 days after irradiation; The IFN-γ concentration in serum of the mice in gene-radiotherapy group were higher than that in reconstruction plasmids group and control group 1 and 3 days after irradiation; NK cytotoxic activity, IL-2 and IFN-γ secretion activity of the mice in gene-radiotherapy group were higher than that in 20 Gy X-ray irradiation group 15 days after irradiation.Conclusions: The anti-tumor effect of repeated low dose gene-radiotherapy was better than that of high dose gene-radiotherapy for only once. By inducing higher expression of IFN-γ gene in the tumors, pEgr-IFN-γ gene-radiotherapy could increase the concentration of IFN-γ in serum, and therefore the body’s immunologic function and anti tumor ability were enhanced.
Objective：To study the anti-tumor effect of repeated low dose pEgr-IFN-γ gene-radiotherapy in mice bearing melanoma and its immunologic mechanism. Methods:The pEgr-IFN-γ plasmids packed by liposome were injected locally into tumors, and then the tumors was irradiated by X-ray 36 hours later. The tumor growth rate at different time and mean survival period of the mice were observed. The IFN-γ mRNA level in tumors was detected by RT-PCR 3 days after irradiation. The concentration of IFN-γ in serum were detected by ELISA. The immunologic indexes was detected 15 days after irradiation. Results: The tumor growth rate of the mice in gene-radiotherapy group with pEgr-IFN-γ and 5 Gy X-ray irradiation four times were lower significantly than that with pEgr-IFN-γ and 20 Gy X-ray irradiation 9～15 days after irradiation and their mean survival period were longer. The IFN-γ mRNA level in tumors of the mice in gene-radiotherapy group were higher than that in reconstruction plasmids group significantly 3 days after irradiation; The IFN-γ concentration in serum of the mice in gene-radiotherapy group were higher than that in reconstruction plasmids group and control group 1 and 3 days after irradiation; NK cytotoxic activity, IL-2 and IFN-γ secretion activity of the mice in gene-radiotherapy group were higher than that in 20 Gy X-ray irradiation group 15 days after irradiation.Conclusions: The anti-tumor effect of repeated low dose gene-radiotherapy was better than that of high dose gene-radiotherapy for only once. By inducing higher expression of IFN-γ gene in the tumors, pEgr-IFN-γ gene-radiotherapy could increase the concentration of IFN-γ in serum, and therefore the body’s immunologic function and anti tumor ability were enhanced.
2004, 11(3):204-207.
DOI: 10.3872/j.issn.1007-385X.2004.3.012
Abstract:
Objective: To study the effects of prokaryotically expressed recombinant human NDRG2 on tumor cell lines. MethodsRecombinant human NDRG2′s inhibitory effects on cell lines 7901 and HHCC at a range of protein concentrations were measured by MTT colorimeteric assay and cell counting against control groups. In the present study, the cell cycle changes were assessed by flow cytometry, and in vivo tumor inhibition study was performed on nude mice. Results: Recombinant human NDRG2 exerted strong inhibitory effects on tumor cell growth, and started to significantly suppress tumor cell growth at the concentration of about 50 μg/ml. Flow cytometry analysis indicated that such inhibition triggered changes to the G1 phase in the tumor cell cycle. In addition, the growth of the tumor xenografts in nude mice was signficantly suppressed in the in vivo study. Conclusion: Recombinant human NDRG2 exhibits cdertain inhibitory effects on tumor cell growth both in vivo and in vivo, which presents an interesting finding that will offers us insight into the cancer treatment.
Objective: To study the effects of prokaryotically expressed recombinant human NDRG2 on tumor cell lines. MethodsRecombinant human NDRG2′s inhibitory effects on cell lines 7901 and HHCC at a range of protein concentrations were measured by MTT colorimeteric assay and cell counting against control groups. In the present study, the cell cycle changes were assessed by flow cytometry, and in vivo tumor inhibition study was performed on nude mice. Results: Recombinant human NDRG2 exerted strong inhibitory effects on tumor cell growth, and started to significantly suppress tumor cell growth at the concentration of about 50 μg/ml. Flow cytometry analysis indicated that such inhibition triggered changes to the G1 phase in the tumor cell cycle. In addition, the growth of the tumor xenografts in nude mice was signficantly suppressed in the in vivo study. Conclusion: Recombinant human NDRG2 exhibits cdertain inhibitory effects on tumor cell growth both in vivo and in vivo, which presents an interesting finding that will offers us insight into the cancer treatment.
ZHANG Jing-min
,
JIN Ning-yi
,
MI Zhi-qiang
,
ZHANG Hong-gui
,
LI Xiao
,
LIAN Hai
,
SUN Ying-chun
,
AN Ru-guo
,
Takahashi
,
M
2004, 11(3):208-211.
DOI: 10.3872/j.issn.1007-385X.2004.3.013
Abstract:
Objective: To investigate the distribution of RFP( Ret Finger Protein) in Human, mouse normal tissues and human esophageal squamous cell carcinoma. Methods: Real-time Quantitative RT-PCR with 18SrRNA as the internal standard was used to detect the expression of RFP in human normal tissues such as cervical tissue, endometrial tissue, testis tissue, gastric tissue, esophageal tissue, brain tissue, human esophageal squamous cell carcinoma and mouse normal tissues such as lung tissue, kidney tissue, uterus tissue, ovary tissue, thymus tissue, heart tissue, muscle tissue, testis tissue, skin tissue, esophageal tissue, spleen tissue, liver tissue, colon tissue, intestinum tissue. Results: The highest level of RFP mRNA was only present in testis tissue among human and mouse normal tissues. In addition, the up-regulation of RFP protein was observed in 13 out of 20 human esophageal squamous cell carcinoma tissues, but only the upregulation of RFP in 9 out of 20 is significant（P<0.05）. Conclusions:Regarding to the expression of RFP limited to testis among human and mouse normal tissues and the up-regulation of RFP in 13 out of 20 human esophageal squamous cell carcinoma tissues, the RFP can be used as a potential molecular target for the development of new therapeutics for human esophageal squamous cell carcinoma.
Objective: To investigate the distribution of RFP( Ret Finger Protein) in Human, mouse normal tissues and human esophageal squamous cell carcinoma. Methods: Real-time Quantitative RT-PCR with 18SrRNA as the internal standard was used to detect the expression of RFP in human normal tissues such as cervical tissue, endometrial tissue, testis tissue, gastric tissue, esophageal tissue, brain tissue, human esophageal squamous cell carcinoma and mouse normal tissues such as lung tissue, kidney tissue, uterus tissue, ovary tissue, thymus tissue, heart tissue, muscle tissue, testis tissue, skin tissue, esophageal tissue, spleen tissue, liver tissue, colon tissue, intestinum tissue. Results: The highest level of RFP mRNA was only present in testis tissue among human and mouse normal tissues. In addition, the up-regulation of RFP protein was observed in 13 out of 20 human esophageal squamous cell carcinoma tissues, but only the upregulation of RFP in 9 out of 20 is significant（P<0.05）. Conclusions:Regarding to the expression of RFP limited to testis among human and mouse normal tissues and the up-regulation of RFP in 13 out of 20 human esophageal squamous cell carcinoma tissues, the RFP can be used as a potential molecular target for the development of new therapeutics for human esophageal squamous cell carcinoma.
2004, 11(3):212-214.
DOI: 10.3872/j.issn.1007-385X.2004.3.014
Abstract:
Objective：To investigate the effect of helper T cells (Th1 and Th2) and its important significance in judging prognosis of patients with advanced lung cancer. Methods： Interferon-γ (INF-γ) and interleukin-4 (IL-4) in plasma and pleural effusion were detected by enzyme-linked immunoassay in patients with advanced lung cancer, IFN-γ and IL-4 reflected activity of Th1 and Th2 respectively. Results： The activity of Th1 in patients with advanced lung cancer was lower than that in normal persons in peripheral blood. For non-outstanding curative effect or progressive state of illness, the activity of Th1 in patients of above 1 year survival time decreased in post-treatment than in pretreatment, the activity of Th2 increased in post-treatment. Conclusion： Activity of Helper T cells (Th1, Th2 ) could be an important marker to diagnose lung cancer and judge prognosis in patients with advanced lung cancer.
Objective：To investigate the effect of helper T cells (Th1 and Th2) and its important significance in judging prognosis of patients with advanced lung cancer. Methods： Interferon-γ (INF-γ) and interleukin-4 (IL-4) in plasma and pleural effusion were detected by enzyme-linked immunoassay in patients with advanced lung cancer, IFN-γ and IL-4 reflected activity of Th1 and Th2 respectively. Results： The activity of Th1 in patients with advanced lung cancer was lower than that in normal persons in peripheral blood. For non-outstanding curative effect or progressive state of illness, the activity of Th1 in patients of above 1 year survival time decreased in post-treatment than in pretreatment, the activity of Th2 increased in post-treatment. Conclusion： Activity of Helper T cells (Th1, Th2 ) could be an important marker to diagnose lung cancer and judge prognosis in patients with advanced lung cancer.
联系方式
- 《中国肿瘤生物治疗杂志》
- 1994年创刊
- 主办单位：Chinese Society of Immunology, Chinese Anti-cancer Association
- 邮编：200433
- 电话：021-81871002-22
- 电子邮箱：cjcb@biother.cn
- 网址:http://www.biother.cn
- 刊号：ISSN 1007-385X
- CN 31-1725/R
- 国内定价： ￥20元/册
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Chinese Journal Of Cancer Biotheray ® 2025 All Rights Reserved
Supported by:Beijing E-Tiller Technology Development Co., Ltd.
Chinese Journal Of Cancer Biotheray ® 2025 All Rights Reserved
Supported by:Beijing E-Tiller Technology Development Co., Ltd.