Mobile website
Volume 10,Issue 1,2003 Table of Contents
2003, 10(1):1-4.
DOI: 10.3872/j.issn.1007-385X.2003.1.001
Abstract:
基因治疗作为一种新型的治疗方法 ,其首要的技术问题就是实现外源基因的输送。在基因输送的研究领域中人们一直在探寻一种安全、高效的基因运输载体。许多种合成的载体被研究作为可能的基因传输工具。这些合成的载体包括带正电的多肽 ,阳离子脂质体和阳离子多聚物等 ,与病毒载体相比 ,这些载体具有制备方法简单 ,免疫原性较低 ,安全低毒等优点。但它们的基因转染效率都低于重组病毒载体 ,如腺病毒。ＰＡＭＡＭ(Ｐｏｌｙａｍｉｄｏａｍｉｎｅ)Ｄｅｎｄｒｉｍｅｒ是最早被合成、定性并商业化的一种纳米级球状多聚物。其大小、形状及功能基团的…
基因治疗作为一种新型的治疗方法 ,其首要的技术问题就是实现外源基因的输送。在基因输送的研究领域中人们一直在探寻一种安全、高效的基因运输载体。许多种合成的载体被研究作为可能的基因传输工具。这些合成的载体包括带正电的多肽 ,阳离子脂质体和阳离子多聚物等 ,与病毒载体相比 ,这些载体具有制备方法简单 ,免疫原性较低 ,安全低毒等优点。但它们的基因转染效率都低于重组病毒载体 ,如腺病毒。ＰＡＭＡＭ(Ｐｏｌｙａｍｉｄｏａｍｉｎｅ)Ｄｅｎｄｒｉｍｅｒ是最早被合成、定性并商业化的一种纳米级球状多聚物。其大小、形状及功能基团的…
2003, 10(1):5-8.
DOI: 10.3872/j.issn.1007-385X.2003.1.002
Abstract:
Objective: To investigate whether carcino-embryonic antigen(CEA) promotor determines the specific expression of E. coli. cytotosine deaminase(EC-CD) gene and the specific cytoxicity in CEA-producing colorectal carcinoma cells. Methods:Two recombinant adenovirus vectors, AdCEACD containing EC-CD gene controlled under CEA promotor and AdCMVCD containing cytomegalovirus(CMV) promotor and EC-CD gene, were constructed. The expression of EC-CD gene in cells was tested with RT-PCR and cytotoxic effects were assayed with MTT method. Results：The CEA-producing cells (human colorectal carcinoma cell line Lovo cell) showed EC-CD mRNA expression and became sensitive to 5-fluorocytosine (5-FC) after infection with AdCEACD and AdCMVCD respectively, The CEA-nonproducing cells(Hela cell) expressed EC-CD mRNA and were sensitive to 5-FC after infection with AdCMVCD, but had no expression of EC CD mRNA and no cytotoxic effect after infection with AdCEACD. Conclusion：CEA promotor could control the expression of EC-CD gene in CEA-positive colorectal carcinoma cells specifically in vitro.
Objective: To investigate whether carcino-embryonic antigen(CEA) promotor determines the specific expression of E. coli. cytotosine deaminase(EC-CD) gene and the specific cytoxicity in CEA-producing colorectal carcinoma cells. Methods:Two recombinant adenovirus vectors, AdCEACD containing EC-CD gene controlled under CEA promotor and AdCMVCD containing cytomegalovirus(CMV) promotor and EC-CD gene, were constructed. The expression of EC-CD gene in cells was tested with RT-PCR and cytotoxic effects were assayed with MTT method. Results：The CEA-producing cells (human colorectal carcinoma cell line Lovo cell) showed EC-CD mRNA expression and became sensitive to 5-fluorocytosine (5-FC) after infection with AdCEACD and AdCMVCD respectively, The CEA-nonproducing cells(Hela cell) expressed EC-CD mRNA and were sensitive to 5-FC after infection with AdCMVCD, but had no expression of EC CD mRNA and no cytotoxic effect after infection with AdCEACD. Conclusion：CEA promotor could control the expression of EC-CD gene in CEA-positive colorectal carcinoma cells specifically in vitro.
2003, 10(1):9-12.
DOI: 10.3872/j.issn.1007-385X.2003.1.003
Abstract:
Objective: To investigate the inhibitory effects of Ad-K5 on the proliferation of endothelial cells and the growth of mammary cancer.Methods: Kringle 5 of human plasminogen was constructed by PCR, and then cloned to plasmid pCA13. Recombinant adenovirus Ad-K5 was obtained through homologous recombination in 293 cells. The inhibition effect of Ad-K5 on cell proliferation was observed in vitro. The tumor size was measured at different intervals to observe the antitumoral effect of Ad-K5 in vivo.Results:The mRNA expression of K5 gene was detected in Ad-K5 infected B-Cap-37 cells. Ad-K5 inhibited the growth of ECV-304 cells, but had almost no effects on B-Cap-37 cells. It seemed that Ad-K5 could inhibit endothelial cells but not kill the cancer cells directly. Ad-K5 could also inhibit the tumor growth in vivo.Conclusion: Ad-K5 could inhibit the growth of solid mammary tumor.
Objective: To investigate the inhibitory effects of Ad-K5 on the proliferation of endothelial cells and the growth of mammary cancer.Methods: Kringle 5 of human plasminogen was constructed by PCR, and then cloned to plasmid pCA13. Recombinant adenovirus Ad-K5 was obtained through homologous recombination in 293 cells. The inhibition effect of Ad-K5 on cell proliferation was observed in vitro. The tumor size was measured at different intervals to observe the antitumoral effect of Ad-K5 in vivo.Results:The mRNA expression of K5 gene was detected in Ad-K5 infected B-Cap-37 cells. Ad-K5 inhibited the growth of ECV-304 cells, but had almost no effects on B-Cap-37 cells. It seemed that Ad-K5 could inhibit endothelial cells but not kill the cancer cells directly. Ad-K5 could also inhibit the tumor growth in vivo.Conclusion: Ad-K5 could inhibit the growth of solid mammary tumor.
2003, 10(1):13-16.
DOI: 10.3872/j.issn.1007-385X.2003.1.004
Abstract:
Objective:To evaluate the inhititory effects of the recombinant adenovirus vector expressing melittin gene under the control of human α-fetoprotein (AFP).Methods:The melittin gene was inserted into adenoviral vector through a bacterial homologous recombinant system, then the recombinant adenoviral vector was transfected to 293 cells to generate the desired recombinant adenoviruses. AFP-positive or AFP-negative hepatocarcinoma cells or hepatic cells were infected respectively with the recombinant adenoviruses and the proliferative inhibition was evaluated by MTT method.Results:Recombinant adenovirus containing melittin gene and AFP promoter was constructed successfully. When infected with the recombinant adenoviruses the inhibition of proliferative was observed in AFP-positive hepatocarcinoma cells, but not in AFP-negative hepatocarcinoma cells. Conclusions: Recombinant adenovirus expressing melittin gene under the control of human AFP promoter could specifically inhibit the proliferation of AFP-positive hepatocarcinoma cells.
Objective:To evaluate the inhititory effects of the recombinant adenovirus vector expressing melittin gene under the control of human α-fetoprotein (AFP).Methods:The melittin gene was inserted into adenoviral vector through a bacterial homologous recombinant system, then the recombinant adenoviral vector was transfected to 293 cells to generate the desired recombinant adenoviruses. AFP-positive or AFP-negative hepatocarcinoma cells or hepatic cells were infected respectively with the recombinant adenoviruses and the proliferative inhibition was evaluated by MTT method.Results:Recombinant adenovirus containing melittin gene and AFP promoter was constructed successfully. When infected with the recombinant adenoviruses the inhibition of proliferative was observed in AFP-positive hepatocarcinoma cells, but not in AFP-negative hepatocarcinoma cells. Conclusions: Recombinant adenovirus expressing melittin gene under the control of human AFP promoter could specifically inhibit the proliferation of AFP-positive hepatocarcinoma cells.
2003, 10(1):17-20.
DOI: 10.3872/j.issn.1007-385X.2003.1.005
Abstract:
Objective: Methylotropic yeast pichia pastoris system was used to express recombinant human soluble 4-1BBL protein with biological activity.Methods:According to the nuclear acid sequence coding human soluble 4-1BBL, we cloned the genes with PCR from XG-4-1BBL transfection cell line,then the gene fragment for extracellar domain was subcloned into the PUCm-T vector and sequence of s4-1BBL cDNA was confirmed by sequencing. The s4-1BBL gene was inserted into the pPICZαA , which was transformed into Pichia pastoris GS115 by linearized electroportion.The recombinant protein was identified by the assay of SDS-PAGE and Western-blot. Costimulating activity of rhs4-1BBL on T cell proliferation in vitro was evidenced by 3H-TdR incorporation assay.Results: The s4-1BBL cDNA was successfully obtained and insected into pPICZαA. The protein molecular weight of hs4-1BBL in the yeast supernamant was about 21 kD by SDS-PAGE analyses,and the specificitity was identified by western blot. Finally, rhs4-1BBL protein could costimulate the proliferation of T cells in vitro.Conclusion: The rhs4-1BBL protein was efficiently expressed in Pichia pastoris (GS115)and showed natural biological activities. And it may provide a valuable materials for further study of 4-1BB/4-1BBL.
Objective: Methylotropic yeast pichia pastoris system was used to express recombinant human soluble 4-1BBL protein with biological activity.Methods:According to the nuclear acid sequence coding human soluble 4-1BBL, we cloned the genes with PCR from XG-4-1BBL transfection cell line,then the gene fragment for extracellar domain was subcloned into the PUCm-T vector and sequence of s4-1BBL cDNA was confirmed by sequencing. The s4-1BBL gene was inserted into the pPICZαA , which was transformed into Pichia pastoris GS115 by linearized electroportion.The recombinant protein was identified by the assay of SDS-PAGE and Western-blot. Costimulating activity of rhs4-1BBL on T cell proliferation in vitro was evidenced by 3H-TdR incorporation assay.Results: The s4-1BBL cDNA was successfully obtained and insected into pPICZαA. The protein molecular weight of hs4-1BBL in the yeast supernamant was about 21 kD by SDS-PAGE analyses,and the specificitity was identified by western blot. Finally, rhs4-1BBL protein could costimulate the proliferation of T cells in vitro.Conclusion: The rhs4-1BBL protein was efficiently expressed in Pichia pastoris (GS115)and showed natural biological activities. And it may provide a valuable materials for further study of 4-1BB/4-1BBL.
2003, 10(1):21-24.
DOI: 10.3872/j.issn.1007-385X.2003.1.006
Abstract:
Objective:To study the characterization of NK cell line transfected with hSCF gene.Methods:pcDNA3-hSCF was transfected into NK-92 cell line with LipofectAMINE ,RT-PCR was used to identify NK-92-hSCF cell which express hSCF and the activity was assayed by TF-1 cell line. CD3, CD16 and CD56 molecules were tested by FACS. Results: We established NK-92-hSCF cell line which express hSCF steadily, and its proliferation increased compared with parental cell line when incubated with rhIL-15 or rhIL-2. Conclusion: The characterization of NK-92 could be changed by transfecting with hSCF gene, and it′s proliferation was improved, The gene transfection of NK-92 cell made it suitable for clinical applications.
Objective:To study the characterization of NK cell line transfected with hSCF gene.Methods:pcDNA3-hSCF was transfected into NK-92 cell line with LipofectAMINE ,RT-PCR was used to identify NK-92-hSCF cell which express hSCF and the activity was assayed by TF-1 cell line. CD3, CD16 and CD56 molecules were tested by FACS. Results: We established NK-92-hSCF cell line which express hSCF steadily, and its proliferation increased compared with parental cell line when incubated with rhIL-15 or rhIL-2. Conclusion: The characterization of NK-92 could be changed by transfecting with hSCF gene, and it′s proliferation was improved, The gene transfection of NK-92 cell made it suitable for clinical applications.
2003, 10(1):25-29.
DOI: 10.3872/j.issn.1007-385X.2003.1.007
Abstract:
Objective: To clone and express functional part of mutant M-CSF (muM-CSF) from human leukemic cell line J6-1 and investigate its Kd for dissociation and biological activity on the proliferation of J6-1.Method: Functional part of muM-CSF was cloned by RT-PCR and inserted into pET32c(+) and expressed in E.coli BL21trxB (DE3). The recombinant protein was purified through Ni2+ affinity column and antibody linked affinity column. ELISA was performed to define the Kd of the muM-CSF to its receptor. Colony formation assay was performed to test its effects on the proliferation of J6-1. Results: The protein was purified and its Kd to the receptor was 3.7 nmol/L. muM-CSF showed elevated proliferation-stimulating potential than normal M-CSF. Conclusion: muM-CSF could be expressed in the pET32c(+), BL21 system and the muM-CSF showed elevated proliferation promoting ability as to normal M-CSF.
Objective: To clone and express functional part of mutant M-CSF (muM-CSF) from human leukemic cell line J6-1 and investigate its Kd for dissociation and biological activity on the proliferation of J6-1.Method: Functional part of muM-CSF was cloned by RT-PCR and inserted into pET32c(+) and expressed in E.coli BL21trxB (DE3). The recombinant protein was purified through Ni2+ affinity column and antibody linked affinity column. ELISA was performed to define the Kd of the muM-CSF to its receptor. Colony formation assay was performed to test its effects on the proliferation of J6-1. Results: The protein was purified and its Kd to the receptor was 3.7 nmol/L. muM-CSF showed elevated proliferation-stimulating potential than normal M-CSF. Conclusion: muM-CSF could be expressed in the pET32c(+), BL21 system and the muM-CSF showed elevated proliferation promoting ability as to normal M-CSF.
2003, 10(1):30-33.
DOI: 10.3872/j.issn.1007-385X.2003.1.008
Abstract:
Objective:To explore the ganciclovir（GCV） therapy on the severe combined immunodeficiency (SCID) breast cancer mice after HSVtk gene and Tet-On regulatory gene expression induced by doxycycline（Dox）. Methods:Co-transfections of pRevTRE/HSVtk and pRevTet-On were performed on MCF-7 cell line by using modified calcium phosphate precipitattion method respectively. After MCF/TRE/tk/Tet-On cell line was established，cells were inoculated into the SCID mice to form the human breast cancer in SCID mice. Regimen (GCV, GCV＋DOX, Normal Saline) was injected introperitoneally in SCID mice 15 days. The volume of tumor was measured and the tumor tissues were examined pathologically. HSVtk gene expression was analyzed by RT-PCR. Results:The human breast cancer in SCID mice was formed successfully by inoculated MCF/TRE/tk/Tet-On cells into the female SCID mice. In the GCV＋DOX-treated group, volume of tumors was shrank remarkably after 15 days' treatment and tumor growth were retarded compared with the control groups. There was significant difference between the GCV＋DOX-treated group and the control groups (P<0.05). Infiltrating cells were observed in tumors that injected with doxycycline followed by GCV treatment and cells were profoundly damaged stained with hematoxylin and eosin. The expression of tk gene in SCID mice tumors was also obeserved by RT-PCR analysis.Conclusions:The human breast cancer in SCID mice was formed successfully by implanted MCF/TRE/tk/Tet-On cells. The growth of tumor could be shrank remarkably with GCV treatment in our SCID mice under the doxycycline induction.
Objective:To explore the ganciclovir（GCV） therapy on the severe combined immunodeficiency (SCID) breast cancer mice after HSVtk gene and Tet-On regulatory gene expression induced by doxycycline（Dox）. Methods:Co-transfections of pRevTRE/HSVtk and pRevTet-On were performed on MCF-7 cell line by using modified calcium phosphate precipitattion method respectively. After MCF/TRE/tk/Tet-On cell line was established，cells were inoculated into the SCID mice to form the human breast cancer in SCID mice. Regimen (GCV, GCV＋DOX, Normal Saline) was injected introperitoneally in SCID mice 15 days. The volume of tumor was measured and the tumor tissues were examined pathologically. HSVtk gene expression was analyzed by RT-PCR. Results:The human breast cancer in SCID mice was formed successfully by inoculated MCF/TRE/tk/Tet-On cells into the female SCID mice. In the GCV＋DOX-treated group, volume of tumors was shrank remarkably after 15 days' treatment and tumor growth were retarded compared with the control groups. There was significant difference between the GCV＋DOX-treated group and the control groups (P<0.05). Infiltrating cells were observed in tumors that injected with doxycycline followed by GCV treatment and cells were profoundly damaged stained with hematoxylin and eosin. The expression of tk gene in SCID mice tumors was also obeserved by RT-PCR analysis.Conclusions:The human breast cancer in SCID mice was formed successfully by implanted MCF/TRE/tk/Tet-On cells. The growth of tumor could be shrank remarkably with GCV treatment in our SCID mice under the doxycycline induction.
2003, 10(1):34-38.
DOI: 10.3872/j.issn.1007-385X.2003.1.009
Abstract:
Objective:Survived from Anoikis is one important reason for maliganant tumor performing matastasis and local invasion. The present study focused on the mechanism and signal transduction of osteosarcoma anoikis induced by human interferon (IFN) α2a.Methods:Interferon-α2a induced anoikis was detected on a non-attachment culture model. The osteosarcoma anoikis was detected by terminal deoxynucleotidyl transferase-mediated dUTP nick and labeling (TUNEL) method. Osteosarcoma cells apoptosis and integrin receptors expression were detected by flow cytometry. The cysteine aspartate-specific proteases (caspase) signal transduction was detected by cleavage of synthethic substractes. Results: Human IFN-α2a induced the anoikis apoptosis of osteosarcoma cell MG- 63, with concomitant upregulation of caspase-8 and caspase-3 activity. The expression of integrin receptor α and αv was not influenced evidently.Conclusion: The present results suggested that human IFN-α2a decreased the metastasis of osteosarcoma by inducing the anoikis of osteosarcoma cell MG- 63. Our results also indicated that caspase signal transduction might regulate this anoikis apoptosis in MG- 63.
Objective:Survived from Anoikis is one important reason for maliganant tumor performing matastasis and local invasion. The present study focused on the mechanism and signal transduction of osteosarcoma anoikis induced by human interferon (IFN) α2a.Methods:Interferon-α2a induced anoikis was detected on a non-attachment culture model. The osteosarcoma anoikis was detected by terminal deoxynucleotidyl transferase-mediated dUTP nick and labeling (TUNEL) method. Osteosarcoma cells apoptosis and integrin receptors expression were detected by flow cytometry. The cysteine aspartate-specific proteases (caspase) signal transduction was detected by cleavage of synthethic substractes. Results: Human IFN-α2a induced the anoikis apoptosis of osteosarcoma cell MG- 63, with concomitant upregulation of caspase-8 and caspase-3 activity. The expression of integrin receptor α and αv was not influenced evidently.Conclusion: The present results suggested that human IFN-α2a decreased the metastasis of osteosarcoma by inducing the anoikis of osteosarcoma cell MG- 63. Our results also indicated that caspase signal transduction might regulate this anoikis apoptosis in MG- 63.
2003, 10(1):39-41.
DOI: 10.3872/j.issn.1007-385X.2003.1.010
Abstract:
Objective:To study the methylation state within MAGE-1 B′B promoter in gastric carcinoma and the association between demethylation and pathological differentiation, the association between demethylation and clinical stage. Methods: Using methylation-sensitive restriction analysis followed by polymerase chain reaction (PCR)，we studied 80 specimen that were obtained from surgical samples (including 40 gastric carcinoma and 40 matched adjacent normal gastric mucosae)．Results: An demethylation state was identified in DNA from gastric carcinoma specimens．The demethvlation rate is 25％(10／40)．In contrast，no demethylation state was identified in DNA from matched adjacent normal gastric mucosae. The differences were Significant statistically. ln our study, the demethylation in poorly, moderately, and well differentiated glandulous-cell carcinoma were detected at frequencies of 50％,18.7% and 8.3％,respectively, The differences were significant statistically （P<0.05）． The demethylation rate in early, 1ate tumor stage was 16.7% and 286％，respectively. The differences were significant statistically(P<0.05). Conclusions: The demethylation in MAGE-1 B′B promoter is anomalous in gastric carcinoma, and maybe it is related to the expression of MAGE-1 gene in gastric carcinoma. The demethylation rate is related to the pathological and disease stages differentiation.
Objective:To study the methylation state within MAGE-1 B′B promoter in gastric carcinoma and the association between demethylation and pathological differentiation, the association between demethylation and clinical stage. Methods: Using methylation-sensitive restriction analysis followed by polymerase chain reaction (PCR)，we studied 80 specimen that were obtained from surgical samples (including 40 gastric carcinoma and 40 matched adjacent normal gastric mucosae)．Results: An demethylation state was identified in DNA from gastric carcinoma specimens．The demethvlation rate is 25％(10／40)．In contrast，no demethylation state was identified in DNA from matched adjacent normal gastric mucosae. The differences were Significant statistically. ln our study, the demethylation in poorly, moderately, and well differentiated glandulous-cell carcinoma were detected at frequencies of 50％,18.7% and 8.3％,respectively, The differences were significant statistically （P<0.05）． The demethylation rate in early, 1ate tumor stage was 16.7% and 286％，respectively. The differences were significant statistically(P<0.05). Conclusions: The demethylation in MAGE-1 B′B promoter is anomalous in gastric carcinoma, and maybe it is related to the expression of MAGE-1 gene in gastric carcinoma. The demethylation rate is related to the pathological and disease stages differentiation.
2003, 10(1):42-47.
DOI: 10.3872/j.issn.1007-385X.2003.1.011
Abstract:
Objective:To explore the inhibition effects of ectogenic wild-type p53 cDNA(Ad-wtp53) on colorectal carcinoma cell lines with different p53 gene status and search for the role of wild type p53 tumor suppressor gene in occurrenc and progress of malignant tumor.Methods:MTT process was taken to choose optimal transfection titre. Three kinds of cell lines(p53 gene deletion, mutation and nomal) were transferred by Ad-wtp53 in optimal titre. The inhibition effects of these cell lines were observed and compared. Results:The best titre is 1000 MOI and p53 gene deletion cell line (THC-8908) shew the highest sensitivity. G1-S transition period blocking effects occurred in all cell lines and G2-M phase regulation effects were not coincidence in three colorectal cell lines. Conclusions:Recombinant adenovirus-mediated wild type p53 gene observably inhibited colorectal carcinoma cell lines growth and proliferation, blocked cell cycle in G0/G1 phase and displayed obvious different actions on G2-M phase among cell lines with different p53 status.
Objective:To explore the inhibition effects of ectogenic wild-type p53 cDNA(Ad-wtp53) on colorectal carcinoma cell lines with different p53 gene status and search for the role of wild type p53 tumor suppressor gene in occurrenc and progress of malignant tumor.Methods:MTT process was taken to choose optimal transfection titre. Three kinds of cell lines(p53 gene deletion, mutation and nomal) were transferred by Ad-wtp53 in optimal titre. The inhibition effects of these cell lines were observed and compared. Results:The best titre is 1000 MOI and p53 gene deletion cell line (THC-8908) shew the highest sensitivity. G1-S transition period blocking effects occurred in all cell lines and G2-M phase regulation effects were not coincidence in three colorectal cell lines. Conclusions:Recombinant adenovirus-mediated wild type p53 gene observably inhibited colorectal carcinoma cell lines growth and proliferation, blocked cell cycle in G0/G1 phase and displayed obvious different actions on G2-M phase among cell lines with different p53 status.
12 All Trans Retinoid Acid Inhibit Cell Growth in Human Retinoblastoma Cells Via Phosphorylation of JNK
2003, 10(1):48-50.
DOI: 10.3872/j.issn.1007-385X.2003.1.012
Abstract:
Objective: To investigate all trans retinoid Acid (ATRA) inhibition of cell growth in human retinoblastoma Y79 cells, and its mechanism. Methods:Antiproliferation effects of ATRA on Y79 cells were determined by 3H-thymidine incorporation. Cell cycle analysis was performed by flow cytometry. JNK phosphorylation was analyzed by Western blot analysis.Results: After 36 h treatment with 10-6mol·L-1 ATRA, 3H-thymidine incorporation decreased to 40%, under the same condition, Y79 cells were arrested in G0/G1 and Sub-G1 peak appeared. Curcumin, JNK blocker, blocked the growth inhibition by ATRA. JNK was phosphorylated in 10 to 20 min. Conclusion: JNK-phosphorylating mediated ATRA inhibition of apoptosis in Y79 cells. These results suggested that ATRA might have clinical application for treatment of retinoblastoma.
Objective: To investigate all trans retinoid Acid (ATRA) inhibition of cell growth in human retinoblastoma Y79 cells, and its mechanism. Methods:Antiproliferation effects of ATRA on Y79 cells were determined by 3H-thymidine incorporation. Cell cycle analysis was performed by flow cytometry. JNK phosphorylation was analyzed by Western blot analysis.Results: After 36 h treatment with 10-6mol·L-1 ATRA, 3H-thymidine incorporation decreased to 40%, under the same condition, Y79 cells were arrested in G0/G1 and Sub-G1 peak appeared. Curcumin, JNK blocker, blocked the growth inhibition by ATRA. JNK was phosphorylated in 10 to 20 min. Conclusion: JNK-phosphorylating mediated ATRA inhibition of apoptosis in Y79 cells. These results suggested that ATRA might have clinical application for treatment of retinoblastoma.
2003, 10(1):51-53.
DOI: 10.3872/j.issn.1007-385X.2003.1.013
Abstract:
Objective：To investigate the effects and mechanism of adenovirus-bcl-xs gene on the inhibition of ascites tumor cells and survival rate of nude mice transplanted intraperitoneally with human ovarian carcinoma. Methods：By using of a reproduced adenovirus-bcl-xs gene infected in NUTU-19 cells, we transfered it intraperitoneally to ascites tumor model of human ovarian carcinoma transplanted in nude mice, detected the ascites formation, the survival time and survival rate of nude mice with the human ascites tumor. The weight and toxic-adverse systemically effects of nude mice were observed and the gene expression was detected by immunocellchemistry. The apoptotic cells were quantitatively determined by terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL).Results：The adenovirus-bcl-xs gene had inhibitory potential on NUTU-19 cells. bcl-xs protein was expressed and apoptotic cells were observed. The survival time of nude mice was longer and the survival rate was higher, and the time of ascites formation was retarted. Conclusions：The results suggested that the transfer of adenovirus-bcl-xs gene to the ascites tumour of nude mice with human ovarian carcinoma could improve the survival rate of nude mice and retard the time of ascites formation. It may be a useful method of gene therapy in the treatment of ovarian carcinoma.
Objective：To investigate the effects and mechanism of adenovirus-bcl-xs gene on the inhibition of ascites tumor cells and survival rate of nude mice transplanted intraperitoneally with human ovarian carcinoma. Methods：By using of a reproduced adenovirus-bcl-xs gene infected in NUTU-19 cells, we transfered it intraperitoneally to ascites tumor model of human ovarian carcinoma transplanted in nude mice, detected the ascites formation, the survival time and survival rate of nude mice with the human ascites tumor. The weight and toxic-adverse systemically effects of nude mice were observed and the gene expression was detected by immunocellchemistry. The apoptotic cells were quantitatively determined by terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL).Results：The adenovirus-bcl-xs gene had inhibitory potential on NUTU-19 cells. bcl-xs protein was expressed and apoptotic cells were observed. The survival time of nude mice was longer and the survival rate was higher, and the time of ascites formation was retarted. Conclusions：The results suggested that the transfer of adenovirus-bcl-xs gene to the ascites tumour of nude mice with human ovarian carcinoma could improve the survival rate of nude mice and retard the time of ascites formation. It may be a useful method of gene therapy in the treatment of ovarian carcinoma.
2003, 10(1):54-57.
DOI: 10.3872/j.issn.1007-385X.2003.1.014
Abstract:
Objective: To construct anti-human prostate specific antigen (PSA) single chain Fv antibody (scFv)/human p53 tetramerization domain fusion gene and express fusion protein in HeLa cells.Methods:The human IgG3 upper hinge/human p53 tetramerization domain fusion gene was obtained by recursive polymerase chain reaction (PCR), and was inserted into pUC19 to construct cloning plasmid pUC19/IgG3/p53. The anti-PSA scFv was then cloned into pUC19/IgG3/p53 to construct anti-PSA scFv /human p53 tetramerization domain fusion gene which was then subcloned into the pSecTag2-B expression plasmid. Then the pSecTag2-B plasmids concluding the fusion gene were transfected HeLa cells. The expression products were analyzed by both SDS-PAGE and Western blot, then were purified with Ni2+-NTA superflow affinity chromatography. The binding affinity for PC-3 cells was measured by flow cytometry.Results:The anti-PSA scFv/human p53 tetramerization domain fusion gene consisted of 891bp encoding 297 amino acid residues, and was the same as that reported before. The expression products of the tetrameric anti-PSA scFv, which relative molecular mass (Mr) was about 35 000, were confirmed by SDS-PAGE and Western blot. After purified with Ni2+-NTA superflow affinity chromatography, the tetrameric anti-PSA scFv showed significantly stronger binding to PC-3 cells than scFv.Conclusion: The tetrameric anti-PSA scFv which could bind to PC-3 cells has been successfully gained for the potential use in clinical studies.
Objective: To construct anti-human prostate specific antigen (PSA) single chain Fv antibody (scFv)/human p53 tetramerization domain fusion gene and express fusion protein in HeLa cells.Methods:The human IgG3 upper hinge/human p53 tetramerization domain fusion gene was obtained by recursive polymerase chain reaction (PCR), and was inserted into pUC19 to construct cloning plasmid pUC19/IgG3/p53. The anti-PSA scFv was then cloned into pUC19/IgG3/p53 to construct anti-PSA scFv /human p53 tetramerization domain fusion gene which was then subcloned into the pSecTag2-B expression plasmid. Then the pSecTag2-B plasmids concluding the fusion gene were transfected HeLa cells. The expression products were analyzed by both SDS-PAGE and Western blot, then were purified with Ni2+-NTA superflow affinity chromatography. The binding affinity for PC-3 cells was measured by flow cytometry.Results:The anti-PSA scFv/human p53 tetramerization domain fusion gene consisted of 891bp encoding 297 amino acid residues, and was the same as that reported before. The expression products of the tetrameric anti-PSA scFv, which relative molecular mass (Mr) was about 35 000, were confirmed by SDS-PAGE and Western blot. After purified with Ni2+-NTA superflow affinity chromatography, the tetrameric anti-PSA scFv showed significantly stronger binding to PC-3 cells than scFv.Conclusion: The tetrameric anti-PSA scFv which could bind to PC-3 cells has been successfully gained for the potential use in clinical studies.
2003, 10(1):58-59.
DOI: 10.3872/j.issn.1007-385X.2003.1.015
Abstract:
Objective: To construct anti human prostate specific antigen (PSA) single chain Fv antibody (scFv)/human p53 tetramerization domain fusion gene and express fusion protein in HeLa cells. Methods: The human IgG3 upper hinge/human p53 tetramerization domain fusion gene was obtained by recursive polymerase chain reaction (PCR), and was inserted into pUC19 to construct cloning plasmid pUC19/IgG3/p53. The anti PSA scFv was then cloned into pUC19/IgG3/p53 to construct anti PSA scFv /human p53 tetramerization domain fusion gene which was then subcloned into the pSecTag2 B expression plasmid. Then the pSecTag2 B plasmids concluding the fusion gene were transfected HeLa cells. The expression products were analyzed by both SDS PAGE and Western blot, then were purified with Ni 2+ NTA superflow affinity chromatography. The binding affinity for PC 3 cells was measured by flow cytometry. Results: The anti PSA scFv/human p53 tetramerization domain fusion gene consisted of 891bp encoding 297 amino acid residues, and was the same as that reported before. The expression products of the tetrameric anti PSA scFv, which relative molecular mass (Mr) was about 35 000, were confirmed by SDS PAGE and Western blot. After purified with Ni 2+ NTA superflow affinity chromatography, the tetrameric anti PSA scFv showed significantly stronger binding to PC 3 cells than scFv.Conclusion: The tetrameric anti PSA scFv which could bind to PC 3 cells has been successfully gained for the potential use in clinical studies.
Objective: To construct anti human prostate specific antigen (PSA) single chain Fv antibody (scFv)/human p53 tetramerization domain fusion gene and express fusion protein in HeLa cells. Methods: The human IgG3 upper hinge/human p53 tetramerization domain fusion gene was obtained by recursive polymerase chain reaction (PCR), and was inserted into pUC19 to construct cloning plasmid pUC19/IgG3/p53. The anti PSA scFv was then cloned into pUC19/IgG3/p53 to construct anti PSA scFv /human p53 tetramerization domain fusion gene which was then subcloned into the pSecTag2 B expression plasmid. Then the pSecTag2 B plasmids concluding the fusion gene were transfected HeLa cells. The expression products were analyzed by both SDS PAGE and Western blot, then were purified with Ni 2+ NTA superflow affinity chromatography. The binding affinity for PC 3 cells was measured by flow cytometry. Results: The anti PSA scFv/human p53 tetramerization domain fusion gene consisted of 891bp encoding 297 amino acid residues, and was the same as that reported before. The expression products of the tetrameric anti PSA scFv, which relative molecular mass (Mr) was about 35 000, were confirmed by SDS PAGE and Western blot. After purified with Ni 2+ NTA superflow affinity chromatography, the tetrameric anti PSA scFv showed significantly stronger binding to PC 3 cells than scFv.Conclusion: The tetrameric anti PSA scFv which could bind to PC 3 cells has been successfully gained for the potential use in clinical studies.
2003, 10(1):63-65.
DOI: 10.3872/j.issn.1007-385X.2003.1.017
Abstract:
抗表皮生长因子受体（EGFR）单克隆抗体cetuximab（C 225）特异性与EGFR高亲和力结合,从而阻断表皮生长因子（EGF）、转化生长因子 α（TGF α）与EGFR结合及其引起的细胞增殖。体内外的临床前实验都显示cetuximab通过调节细胞周期、抑制血管生成和转移及促进凋亡等抑制肿瘤，而且可以增加放化疗疗效。Ⅰ,Ⅱ期临床试验显示，药代动力学呈剂量依赖非线性关系，半衰期长，高效低毒，在人体耐受良好。大量临床试验已证实cetuximab单药及联合化疗或联合放疗均取得令人鼓舞的结果。
抗表皮生长因子受体（EGFR）单克隆抗体cetuximab（C 225）特异性与EGFR高亲和力结合,从而阻断表皮生长因子（EGF）、转化生长因子 α（TGF α）与EGFR结合及其引起的细胞增殖。体内外的临床前实验都显示cetuximab通过调节细胞周期、抑制血管生成和转移及促进凋亡等抑制肿瘤，而且可以增加放化疗疗效。Ⅰ,Ⅱ期临床试验显示，药代动力学呈剂量依赖非线性关系，半衰期长，高效低毒，在人体耐受良好。大量临床试验已证实cetuximab单药及联合化疗或联合放疗均取得令人鼓舞的结果。
2003, 10(1):65-67.
DOI: 10.3872/j.issn.1007-385X.2003.1.018
Abstract:
一种能特异性抑制血管内皮细胞生长的抑制因子—内皮抑素（endostatin），可以抑制内皮细胞的增殖和迁移，在体内具有良好的抗肿瘤效果。内皮抑素的抗肿瘤效果可能与诱导凋亡，以及与原肌球蛋白、黏附受体整合蛋白以及基质金属蛋白酶等重要因子有关。
一种能特异性抑制血管内皮细胞生长的抑制因子—内皮抑素（endostatin），可以抑制内皮细胞的增殖和迁移，在体内具有良好的抗肿瘤效果。内皮抑素的抗肿瘤效果可能与诱导凋亡，以及与原肌球蛋白、黏附受体整合蛋白以及基质金属蛋白酶等重要因子有关。
2003, 10(1):68-70.
DOI: 10.3872/j.issn.1007-385X.2003.1.019
Abstract:
RNA干扰能使特异基因沉默，目前RNA干扰研究成为热点，并成功的在哺乳动物细胞和小鼠体内抑制部分基因的表达，本文介绍了目前RNA干扰作用机理、导入技术和应用的研究进展，可以预见RNA干扰技术在基因功能研究、细胞信号转导以及疾病的基因治疗等方面有着良好的应用前景。
RNA干扰能使特异基因沉默，目前RNA干扰研究成为热点，并成功的在哺乳动物细胞和小鼠体内抑制部分基因的表达，本文介绍了目前RNA干扰作用机理、导入技术和应用的研究进展，可以预见RNA干扰技术在基因功能研究、细胞信号转导以及疾病的基因治疗等方面有着良好的应用前景。
2003, 10(1):71-74.
DOI: 10.3872/j.issn.1007-385X.2003.1.020
Abstract:
人乳头瘤病毒（HPV）是由核酸和衣壳蛋白组成的DNA病毒，基因组编码10个开放阅读框（ORF），分为早期区（E区），晚期区（L区）和上游调节区（URR），早期区包含E1，E2，E4，E5，E6及E7 6个早期基因，编码合成蛋白，调控病毒转录，复制和转化；晚期区L1和L2编码病毒的主要和次要衣壳蛋白。高危型HPV感染与宫颈癌有关。高危型HPV E6/E7被证实为转化基因，在相关组织中构成性表达，具有很强的抗原性，被首选用来制备HPV基因疫苗；HPV晚期基因L1的产物具有诱导产生中和抗体及细胞免疫的表位，也是制备基因疫苗的理想候选基因。针对E6，E7和L1等基因序列构建的基因疫苗可同时激活体液免疫和细胞免疫，对宫颈癌有预防和治疗作用。
人乳头瘤病毒（HPV）是由核酸和衣壳蛋白组成的DNA病毒，基因组编码10个开放阅读框（ORF），分为早期区（E区），晚期区（L区）和上游调节区（URR），早期区包含E1，E2，E4，E5，E6及E7 6个早期基因，编码合成蛋白，调控病毒转录，复制和转化；晚期区L1和L2编码病毒的主要和次要衣壳蛋白。高危型HPV感染与宫颈癌有关。高危型HPV E6/E7被证实为转化基因，在相关组织中构成性表达，具有很强的抗原性，被首选用来制备HPV基因疫苗；HPV晚期基因L1的产物具有诱导产生中和抗体及细胞免疫的表位，也是制备基因疫苗的理想候选基因。针对E6，E7和L1等基因序列构建的基因疫苗可同时激活体液免疫和细胞免疫，对宫颈癌有预防和治疗作用。
2003, 10(1):74-76.
DOI: 10.3872/j.issn.1007-385X.2003.1.021
Abstract:
树突状细胞是目前发现的功能最强的抗原递呈细胞，在免疫应答中发挥重要作用。近来，树突状细胞成为肿瘤和免疫学研究的热点，特别是DC瘤苗在肿瘤免疫治疗中的应用前景诱人。本文综述了DC的免疫学活性，抗原摄取和递呈途径，体外诱导扩增及其瘤苗构建和应用的研究进展。
树突状细胞是目前发现的功能最强的抗原递呈细胞，在免疫应答中发挥重要作用。近来，树突状细胞成为肿瘤和免疫学研究的热点，特别是DC瘤苗在肿瘤免疫治疗中的应用前景诱人。本文综述了DC的免疫学活性，抗原摄取和递呈途径，体外诱导扩增及其瘤苗构建和应用的研究进展。
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
您是第位访问者
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.