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Volume 7,Issue 4,2000 Table of Contents
2000, 7(4):243-246.
DOI: 10.3872/j.issn.1007-385X.2000.4.002
Abstract:
:探讨来源于HER2 /neu原癌蛋白的多肽诱导特异性细胞免疫应答及其在体内的抗癌效应。方法 :利用合成的2个具有小鼠H 2Kd 分子结合基序的HER2 /neu肽作为肿瘤排斥抗原 ,在体外刺激小鼠脾淋巴细胞以及皮下免疫小鼠 ,观察淋巴细胞的增殖能力、CTL的杀伤活性以及HER2 /neu肽的抑瘤作用。结果 :HER2 /neu肽在体内和体外对BALB/c小鼠淋巴细胞的增殖都有明显的促进作用。HER2 /neu肽免疫BALB/c小鼠后 ,可以从小鼠淋巴细胞中诱导出肽特异性CTL ,这些CTL可以特异地杀伤HER2 /neu阳性SP2 / 0细胞 ,而对HER2 /neu阴性SP2 / 0细胞却无明显的杀伤活性。SP2 / 0HER2细胞在HER2 /neu肽免疫小鼠体内的生长受到抑制。结论 :研究结果表明HER2 /neu肽可以起到一定的抑瘤保护作用 ,提示 ,肿瘤特异性抗原来源的MHC Ⅰ类分子肽表位作为疫苗进行肿瘤免疫治疗是可行的。
:探讨来源于HER2 /neu原癌蛋白的多肽诱导特异性细胞免疫应答及其在体内的抗癌效应。方法 :利用合成的2个具有小鼠H 2Kd 分子结合基序的HER2 /neu肽作为肿瘤排斥抗原 ,在体外刺激小鼠脾淋巴细胞以及皮下免疫小鼠 ,观察淋巴细胞的增殖能力、CTL的杀伤活性以及HER2 /neu肽的抑瘤作用。结果 :HER2 /neu肽在体内和体外对BALB/c小鼠淋巴细胞的增殖都有明显的促进作用。HER2 /neu肽免疫BALB/c小鼠后 ,可以从小鼠淋巴细胞中诱导出肽特异性CTL ,这些CTL可以特异地杀伤HER2 /neu阳性SP2 / 0细胞 ,而对HER2 /neu阴性SP2 / 0细胞却无明显的杀伤活性。SP2 / 0HER2细胞在HER2 /neu肽免疫小鼠体内的生长受到抑制。结论 :研究结果表明HER2 /neu肽可以起到一定的抑瘤保护作用 ,提示 ,肿瘤特异性抗原来源的MHC Ⅰ类分子肽表位作为疫苗进行肿瘤免疫治疗是可行的。
2000, 7(4):247-250.
DOI: 10.3872/j.issn.1007-385X.2000.4.003
Abstract:
Objective: To investigate the effects of DFMO on the growth characteristics,apoptosis and activity of telomerase of K562 cells.Methods:The growth rate,phase distribution of cell cycel and apoptosis of treated cells with DFMO were detected by cell cuout,morphological assay,FCM analysis and DNA electrophoresis,respectively.The telomerase activiety was examined by telomeric repeat amplification protocol(TRAP).Results: In K562 cells treated with DFMO,the growth rate was markedly retarded.The incrases in G 1-phase cells and decrease in s-phase population and induction of apoptosis were observed.The activity of telomerase of treated cells was inhibited.Conclusion: The growth inhibition and apoptosis induction of K562 cells treated by DFMO were associated with suppressed telomerase activity.It is suggested that inactivation of telomerase in cancer cells conld be one of important events in antitumor molecular mechanism of inhibition of polyamine biosynthesis.
Objective: To investigate the effects of DFMO on the growth characteristics,apoptosis and activity of telomerase of K562 cells.Methods:The growth rate,phase distribution of cell cycel and apoptosis of treated cells with DFMO were detected by cell cuout,morphological assay,FCM analysis and DNA electrophoresis,respectively.The telomerase activiety was examined by telomeric repeat amplification protocol(TRAP).Results: In K562 cells treated with DFMO,the growth rate was markedly retarded.The incrases in G 1-phase cells and decrease in s-phase population and induction of apoptosis were observed.The activity of telomerase of treated cells was inhibited.Conclusion: The growth inhibition and apoptosis induction of K562 cells treated by DFMO were associated with suppressed telomerase activity.It is suggested that inactivation of telomerase in cancer cells conld be one of important events in antitumor molecular mechanism of inhibition of polyamine biosynthesis.
2000, 7(4):251-254.
DOI: 10.3872/j.issn.1007-385X.2000.4.004
Abstract:
研究联合应用bcr abl融合基因硫代磷酸反义寡核苷酸 (Aspo)及c myb基因Aspo对K5 6 2细胞作用效果。 方法 :Aspo与K5 6 2细胞共培养后 ,台盼蓝拒染法计死活细胞数 ;甲基纤维素半固体培养法培养CFU K5 6 2 ;流式细胞仪测定细胞P2 10蛋白表达及细胞凋亡率 ;RT PCR半定量检测细胞bcr ablmRNA ;电镜观察凋亡细胞形态学改变。结果 :倒置显微镜下观察到联合应用两种Aspo时 ,浓度各为 5 μmol/L组K5 6 2细胞仍呈克隆状生长 ,作用 2 4h细胞P2 10蛋白表达明显受抑制 ,表达率 <2 % ;作用 12 0h细胞生长抑制率为 6 1.7% ,P2 10恢复为 2 5 .7% ,凋亡率为 2 2 .5 %。两种浓度各为 10 μmol/L组K5 6 2细胞生长疏散 ,作用 12 0h细胞生长抑制率达 92 .2 % ,P2 10仍未恢复 ,凋亡率为 6 4.3% ,电镜下见细胞呈典型凋亡形态学改变。当两种Aspo浓度各为 5 μmol/L及 10 μmol/L作用 48h时 ,K5 6 2细胞bcr ablmRNA较空白组下降 6 9.2 %～85 3%。结论 :联合应用bcr abl基因Aspo及c myb基因Aspo对K5 6 2细胞的抑制作用具有一定的协同性 ,可增强其抗白血病效果。
研究联合应用bcr abl融合基因硫代磷酸反义寡核苷酸 (Aspo)及c myb基因Aspo对K5 6 2细胞作用效果。 方法 :Aspo与K5 6 2细胞共培养后 ,台盼蓝拒染法计死活细胞数 ;甲基纤维素半固体培养法培养CFU K5 6 2 ;流式细胞仪测定细胞P2 10蛋白表达及细胞凋亡率 ;RT PCR半定量检测细胞bcr ablmRNA ;电镜观察凋亡细胞形态学改变。结果 :倒置显微镜下观察到联合应用两种Aspo时 ,浓度各为 5 μmol/L组K5 6 2细胞仍呈克隆状生长 ,作用 2 4h细胞P2 10蛋白表达明显受抑制 ,表达率 <2 % ;作用 12 0h细胞生长抑制率为 6 1.7% ,P2 10恢复为 2 5 .7% ,凋亡率为 2 2 .5 %。两种浓度各为 10 μmol/L组K5 6 2细胞生长疏散 ,作用 12 0h细胞生长抑制率达 92 .2 % ,P2 10仍未恢复 ,凋亡率为 6 4.3% ,电镜下见细胞呈典型凋亡形态学改变。当两种Aspo浓度各为 5 μmol/L及 10 μmol/L作用 48h时 ,K5 6 2细胞bcr ablmRNA较空白组下降 6 9.2 %～85 3%。结论 :联合应用bcr abl基因Aspo及c myb基因Aspo对K5 6 2细胞的抑制作用具有一定的协同性 ,可增强其抗白血病效果。
2000, 7(4):255-260.
DOI: 10.3872/j.issn.1007-385X.2000.4.005
Abstract:
研究各种白血病细胞膜IL 6R的表达、密度及亲和力 ,为深入探讨对IL 6 /IL 6R系统介导IL 6 PE40外毒素融合蛋白靶向杀伤白血病细胞的敏感性提供可靠依据。方法 :采用放射性配基结合实验 (RBA)并结合Scatchard作图法 ,系统分析IL 6R在 8种极具代表性的人类白血病细胞系U937,HL 6 0 ,KG1,TF1,K5 6 2 ,CEM ,HuT2 8和Raji上的表达密度及亲和力 ,同时应用流式细胞术 (FACS)检测IL 6R的α和 β亚单位蛋白在这些白血病细胞上的表达。结果 :RBA表明 ,粒、单、红白血病细胞系HL 6 0 ,U937,KG1和TF1表面存在着高亲和力IL 6R ,平均IL 6R密度 /细胞分别为 2 5 0 2个 ,2 874个 ,2 319个及 932 9个 ,而淋系白血病细胞系CEM ,HuT2 8和Raji以及慢粒K5 6 2细胞系则未测到IL 6R。FACS显示 ,HL 6 0 ,KG1,U937和TF1均高表达IL 6R的α亚单位蛋白 ,而CEM ,HuT2 8及K5 6 2则不表达IL 6Rα亚单位蛋白 ;IL 6Rβ亚单位蛋白在U937,KG1,TF1,HuT2 8及CEM中呈阳性表达 ,而在HL 6 0 ,Raji和K5 6 2细胞中则为阴性。结论 :鉴于粒、单、红白血病细胞异常高表达IL 6R ,而淋系和慢粒白血病呈阴性表达这一事实 ,提示急非淋白血病可能更适合用重组IL 6 PE40外毒素融合蛋白进行IL 6 /IL 6R介导的靶向治疗。
研究各种白血病细胞膜IL 6R的表达、密度及亲和力 ,为深入探讨对IL 6 /IL 6R系统介导IL 6 PE40外毒素融合蛋白靶向杀伤白血病细胞的敏感性提供可靠依据。方法 :采用放射性配基结合实验 (RBA)并结合Scatchard作图法 ,系统分析IL 6R在 8种极具代表性的人类白血病细胞系U937,HL 6 0 ,KG1,TF1,K5 6 2 ,CEM ,HuT2 8和Raji上的表达密度及亲和力 ,同时应用流式细胞术 (FACS)检测IL 6R的α和 β亚单位蛋白在这些白血病细胞上的表达。结果 :RBA表明 ,粒、单、红白血病细胞系HL 6 0 ,U937,KG1和TF1表面存在着高亲和力IL 6R ,平均IL 6R密度 /细胞分别为 2 5 0 2个 ,2 874个 ,2 319个及 932 9个 ,而淋系白血病细胞系CEM ,HuT2 8和Raji以及慢粒K5 6 2细胞系则未测到IL 6R。FACS显示 ,HL 6 0 ,KG1,U937和TF1均高表达IL 6R的α亚单位蛋白 ,而CEM ,HuT2 8及K5 6 2则不表达IL 6Rα亚单位蛋白 ;IL 6Rβ亚单位蛋白在U937,KG1,TF1,HuT2 8及CEM中呈阳性表达 ,而在HL 6 0 ,Raji和K5 6 2细胞中则为阴性。结论 :鉴于粒、单、红白血病细胞异常高表达IL 6R ,而淋系和慢粒白血病呈阴性表达这一事实 ,提示急非淋白血病可能更适合用重组IL 6 PE40外毒素融合蛋白进行IL 6 /IL 6R介导的靶向治疗。
5 Antitumor Responses Induced by Recombinant Vaccinia Viruses Expressingp53 Antigenic Peptide and B7
2000, 7(4):261-264.
DOI: 10.3872/j.issn.1007-385X.2000.4.006
Abstract:
探索点突变 p5 3抗原肽重组痘苗病毒诱导的抗瘤免疫效应 ,以及B7对之的加强作用 ,为重组抗原肽疫苗用于肿瘤免疫治疗提供实验依据。方法 :选择人 135位Cys→Tyr点突变p5 3为肿瘤相关抗原模型 ,观察包含该点突变p5 3抗原肽p5 312 5～ 14 5的重组痘苗病毒rVV p5 3M 单独和联合应用表达B7的重组痘苗病毒rVV B7诱导的CTL及对荷瘤小鼠的免疫保护和治疗效应。结果 :以rVV p5 3M 经静脉免疫BALB/c小鼠能够诱导以CD8+ T细胞为主的特异性CTL。rVV p5 3M 能够保护部分小鼠免遭致死剂量的肿瘤细胞攻击。以rVV p5 3M 治疗荷瘤小鼠 ,可显著延长小鼠平均存活时间。rVV B7与rVV p5 3M 以 1∶1的比例混合接种可加强rVV p5 3M诱导的抗瘤免疫反应。结论 :以痘苗病毒为载体的p5 3抗原肽疫苗可代替人工合成抗原肽 ,有潜在的临床应用价值 ,共刺激分子B7与肿瘤抗原相结合是一种有效的免疫治疗策略。
探索点突变 p5 3抗原肽重组痘苗病毒诱导的抗瘤免疫效应 ,以及B7对之的加强作用 ,为重组抗原肽疫苗用于肿瘤免疫治疗提供实验依据。方法 :选择人 135位Cys→Tyr点突变p5 3为肿瘤相关抗原模型 ,观察包含该点突变p5 3抗原肽p5 312 5～ 14 5的重组痘苗病毒rVV p5 3M 单独和联合应用表达B7的重组痘苗病毒rVV B7诱导的CTL及对荷瘤小鼠的免疫保护和治疗效应。结果 :以rVV p5 3M 经静脉免疫BALB/c小鼠能够诱导以CD8+ T细胞为主的特异性CTL。rVV p5 3M 能够保护部分小鼠免遭致死剂量的肿瘤细胞攻击。以rVV p5 3M 治疗荷瘤小鼠 ,可显著延长小鼠平均存活时间。rVV B7与rVV p5 3M 以 1∶1的比例混合接种可加强rVV p5 3M诱导的抗瘤免疫反应。结论 :以痘苗病毒为载体的p5 3抗原肽疫苗可代替人工合成抗原肽 ,有潜在的临床应用价值 ,共刺激分子B7与肿瘤抗原相结合是一种有效的免疫治疗策略。
2000, 7(4):265-268.
DOI: 10.3872/j.issn.1007-385X.2000.4.007
Abstract:
Objective: To explore whether gancyclovir (GCV) can inhibit the proliferation and induce the erythro differentiation of the K562 human myeloid leukemia cell line. Methods: K562 cells were cultured with GCV for 4 days to detect cellular changes cloning efficiency, benzidine positive rate, flow cytometry analysis, and telomerase activity. Results: When K562 cells grew in the medium containing GCV, the cellular growth and division were gradually suppressed, growth fracture decreased and further differentiation towards the cell producing hemoglobins was found. Conclusion: GCV can inhibit proliferation and induce erythro differentiation of K562 cells.
Objective: To explore whether gancyclovir (GCV) can inhibit the proliferation and induce the erythro differentiation of the K562 human myeloid leukemia cell line. Methods: K562 cells were cultured with GCV for 4 days to detect cellular changes cloning efficiency, benzidine positive rate, flow cytometry analysis, and telomerase activity. Results: When K562 cells grew in the medium containing GCV, the cellular growth and division were gradually suppressed, growth fracture decreased and further differentiation towards the cell producing hemoglobins was found. Conclusion: GCV can inhibit proliferation and induce erythro differentiation of K562 cells.
2000, 7(4):269-272.
DOI: 10.3872/j.issn.1007-385X.2000.4.008
Abstract:
Objective: To evaluate the inhibition of human endostatin on tumor growth and metastasis of lung adenocarcinoma LA795 in mice. Methods: Recombinant human endostatin was purified from pCX expressed endostatin clones. Plasminogen was purified from outdated human plasma by affinity chromatography, and human angiostatin was produced from human plasminogen digested by elastase and purified by affinity chromatography. LA795 cells were inoculated subcutaneously into the dorsa of T739 mice, and the mice were randomized into 3 groups. From the 10th day, the first group was given 20 mg/kg of recombinant human endostatin s.c. qd, the second was treated daily s.c.of 7.5 mg/kg of human angiostatin, and the third group received daily s.c. with equal volumes of PBS for 14 days. Volumes of the subcutaneous tumors, lung weights, the number of lung surface metastases and mice life span were observed. The results were analyzed by q test. Results: The tumor volumes of both the 1st and the 2nd groups increased slowly. From the 8th day after being treated, the tumor volumes were decreasing. However, in the 3rd group, the tumor volumes increased continuously. The lung weight and the number of lung surface metastases of the 1st and 2nd groups were less than that of the 3rd group. The average survival periods of the 1st and 2nd groups were longer than that of the 3rd group. Conclusion: Human endostatin and angiostatin have strong inhibitory effects both on growth of primary tumor and metastasis of lung adenocarcinoma LA795, and prolongs the survival period of the tumor bearing mice.
Objective: To evaluate the inhibition of human endostatin on tumor growth and metastasis of lung adenocarcinoma LA795 in mice. Methods: Recombinant human endostatin was purified from pCX expressed endostatin clones. Plasminogen was purified from outdated human plasma by affinity chromatography, and human angiostatin was produced from human plasminogen digested by elastase and purified by affinity chromatography. LA795 cells were inoculated subcutaneously into the dorsa of T739 mice, and the mice were randomized into 3 groups. From the 10th day, the first group was given 20 mg/kg of recombinant human endostatin s.c. qd, the second was treated daily s.c.of 7.5 mg/kg of human angiostatin, and the third group received daily s.c. with equal volumes of PBS for 14 days. Volumes of the subcutaneous tumors, lung weights, the number of lung surface metastases and mice life span were observed. The results were analyzed by q test. Results: The tumor volumes of both the 1st and the 2nd groups increased slowly. From the 8th day after being treated, the tumor volumes were decreasing. However, in the 3rd group, the tumor volumes increased continuously. The lung weight and the number of lung surface metastases of the 1st and 2nd groups were less than that of the 3rd group. The average survival periods of the 1st and 2nd groups were longer than that of the 3rd group. Conclusion: Human endostatin and angiostatin have strong inhibitory effects both on growth of primary tumor and metastasis of lung adenocarcinoma LA795, and prolongs the survival period of the tumor bearing mice.
Luo Junsheng
,
Gu Lixue
,
Xi Huanjiu
,
Wei Bingjie
,
Liu Xingbo
,
Qiu Jianwu
,
Zhang Pengfei
,
Shan Hongren
2000, 7(4):273-274.
DOI: 10.3872/j.issn.1007-385X.2000.4.009
Abstract:
Objective: To investigate a new method for improving the therapeutic effect on malignant glioma. Methods: A new type of killer cells, named GS-LAK, was induced by means of costimulating the peripheral ginsenoside(GS) and in-terleukin-2 (IL-2). Comparing with control group-LAK cells, cytotoxicity of GS-LAK cells against malignant glioma cells(BT325) was examined with MTT method. Results: It showed that GS-LAK cells exhibited some advantages over LAK cells in proliferation, cytotoxicity, as well as the utilizing of IL-2. Conclusion: The application of GS-LAK cells might open a new prospect to clinical therapeutic approach to malignant glioma.
Objective: To investigate a new method for improving the therapeutic effect on malignant glioma. Methods: A new type of killer cells, named GS-LAK, was induced by means of costimulating the peripheral ginsenoside(GS) and in-terleukin-2 (IL-2). Comparing with control group-LAK cells, cytotoxicity of GS-LAK cells against malignant glioma cells(BT325) was examined with MTT method. Results: It showed that GS-LAK cells exhibited some advantages over LAK cells in proliferation, cytotoxicity, as well as the utilizing of IL-2. Conclusion: The application of GS-LAK cells might open a new prospect to clinical therapeutic approach to malignant glioma.
2000, 7(4):275-278.
DOI: 10.3872/j.issn.1007-385X.2000.4.010
Abstract:
设计切割ki-rasG12V mRNA的特异性ribozyme(Rz2 17) ,明确其对癌基因ki-rasG12VmRNA的细胞内外切割活性 ,为以ki-rasG12VmRNA为特异性靶分子的基因治疗及癌基因ki-ras的功能研究提拱一种新的途径。方法 :依Symons总结的“锤头结构”原理 ,设计一种能特异性切割ki-rasG12VmRNA的ribozyme ,利用DNA重组技术构建ki-rasG12V外显子 1和ri-bozymeRz2l7的体外转录质粒及ribozymeRz2 17的真核表达质粒 ,体外转录获得ribozymeRz2 17及ki-rasG12V外显子 1mRNA ,在含Mg2 + 溶液中ribozymeRz2 17对其靶RNA分子进行切割。以RT -PCR对转染ribozymeRz2 17真核表达质粒的细胞ki-rasG12VmRNA进行半定量分析。结果 :ki-rasG12V外显子 1体外转录mRNA分子 ,能被ribozymeRz2 17定点切割而野生型ki-ras外显子 1体外转录mRNA则不被切割 ;转染ribozymeRz2 17的胰癌细胞ki-rasG12VmRNA含量减少 ,而转染ri bozymeRz2 17的肝癌细胞其内源性ki-rasmRNA含量无明显变化。结论 :ribozymeRz2 17无论在细胞内外均能剪切突变型ki-rasmRNA(G12V)而且其切割作用为突变型ki-rasG12VmRNA特异性的。
设计切割ki-rasG12V mRNA的特异性ribozyme(Rz2 17) ,明确其对癌基因ki-rasG12VmRNA的细胞内外切割活性 ,为以ki-rasG12VmRNA为特异性靶分子的基因治疗及癌基因ki-ras的功能研究提拱一种新的途径。方法 :依Symons总结的“锤头结构”原理 ,设计一种能特异性切割ki-rasG12VmRNA的ribozyme ,利用DNA重组技术构建ki-rasG12V外显子 1和ri-bozymeRz2l7的体外转录质粒及ribozymeRz2 17的真核表达质粒 ,体外转录获得ribozymeRz2 17及ki-rasG12V外显子 1mRNA ,在含Mg2 + 溶液中ribozymeRz2 17对其靶RNA分子进行切割。以RT -PCR对转染ribozymeRz2 17真核表达质粒的细胞ki-rasG12VmRNA进行半定量分析。结果 :ki-rasG12V外显子 1体外转录mRNA分子 ,能被ribozymeRz2 17定点切割而野生型ki-ras外显子 1体外转录mRNA则不被切割 ;转染ribozymeRz2 17的胰癌细胞ki-rasG12VmRNA含量减少 ,而转染ri bozymeRz2 17的肝癌细胞其内源性ki-rasmRNA含量无明显变化。结论 :ribozymeRz2 17无论在细胞内外均能剪切突变型ki-rasmRNA(G12V)而且其切割作用为突变型ki-rasG12VmRNA特异性的。
2000, 7(4):279-281.
DOI: 10.3872/j.issn.1007-385X.2000.4.011
Abstract:
克隆有特异调控活性的人CD34基因 5′端转录调控序列。方法 :根据已知的CD34抗原基因 5′ 端启动子调控序列 ,自行设计 2对引物 ,用巢式 PCR方法从染色体DNA中成功的克隆 6 6 1bp长度的CD34抗原转录调控序列 (CD34TRS) ,CD34TRS片段插入启动子报告质粒 pEGFP 1,观察重组质粒pCD34EGFP在造血系细胞和非造血系细胞中的调控表达作用。结果 :经限制性内切酶酶切鉴定及DNA序列分析证实 ,克隆的CD34启动子序列与文献报道的顺序基本一致 ,能特异调控EGFP基因在造血细胞系细胞K5 6 2中表达。结论 :所克隆的人CD34分化抗原转录调控序列有特异调控真核基因表达作用 ,本研究为构建造血干细胞特异性表达载体奠定了基础。
克隆有特异调控活性的人CD34基因 5′端转录调控序列。方法 :根据已知的CD34抗原基因 5′ 端启动子调控序列 ,自行设计 2对引物 ,用巢式 PCR方法从染色体DNA中成功的克隆 6 6 1bp长度的CD34抗原转录调控序列 (CD34TRS) ,CD34TRS片段插入启动子报告质粒 pEGFP 1,观察重组质粒pCD34EGFP在造血系细胞和非造血系细胞中的调控表达作用。结果 :经限制性内切酶酶切鉴定及DNA序列分析证实 ,克隆的CD34启动子序列与文献报道的顺序基本一致 ,能特异调控EGFP基因在造血细胞系细胞K5 6 2中表达。结论 :所克隆的人CD34分化抗原转录调控序列有特异调控真核基因表达作用 ,本研究为构建造血干细胞特异性表达载体奠定了基础。
2000, 7(4):282-284.
DOI: 10.3872/j.issn.1007-385X.2000.4.012
Abstract:
To construct the recombinant vaccinia virus of mutant N ras/61 gene and enhance the immunogenecity of mutant N ras/61 protein produced by the recombinant vaccinia virus. Methods: N ras/61 gene was inserted into P1108 and transfected into CV 1 cell infected with vaccinia virus by Cell FECTIN. PCR and Western blot were used to identify the recombinants. Results: We get recombinant vaccinia virus rV N ras/61 by PCR and tk - selecting. The expression of N ras gene was detected by Western blot. Conclusion: This study is a test for studing effective vaccine of mutant N ras/61 gene.The efficacy in vivo of the N ras/61 vaccine in immunotherapy is under investgation.
To construct the recombinant vaccinia virus of mutant N ras/61 gene and enhance the immunogenecity of mutant N ras/61 protein produced by the recombinant vaccinia virus. Methods: N ras/61 gene was inserted into P1108 and transfected into CV 1 cell infected with vaccinia virus by Cell FECTIN. PCR and Western blot were used to identify the recombinants. Results: We get recombinant vaccinia virus rV N ras/61 by PCR and tk - selecting. The expression of N ras gene was detected by Western blot. Conclusion: This study is a test for studing effective vaccine of mutant N ras/61 gene.The efficacy in vivo of the N ras/61 vaccine in immunotherapy is under investgation.
2000, 7(4):285-287.
DOI: 10.3872/j.issn.1007-385X.2000.4.013
Abstract:
Objective: To express recombinant human angiostatin for further application in clinic. Methods: The complete encoding cDNA of human angiostatin was isolated from human embryo liver with RT PCR and expressed in secretory Pichia expression system. Recombinant human angiostatin was purified with heparin sepharose chromatography and its activity was determined in chick embryo chorioallantoic membrane (CAM) and wound healing assays. Results: Expressed in large quantity (yield=5 mg/L) and purified with heparin sepharose, recombinant angiostatin was showed to have a molecular weight of 43 kD in SDS PAGE and potently inhibit angiogenesis and wound healing. Conclusion: Recombinant human angiostatin was expressed efficiently in a biologically active form.
Objective: To express recombinant human angiostatin for further application in clinic. Methods: The complete encoding cDNA of human angiostatin was isolated from human embryo liver with RT PCR and expressed in secretory Pichia expression system. Recombinant human angiostatin was purified with heparin sepharose chromatography and its activity was determined in chick embryo chorioallantoic membrane (CAM) and wound healing assays. Results: Expressed in large quantity (yield=5 mg/L) and purified with heparin sepharose, recombinant angiostatin was showed to have a molecular weight of 43 kD in SDS PAGE and potently inhibit angiogenesis and wound healing. Conclusion: Recombinant human angiostatin was expressed efficiently in a biologically active form.
Wu Guichen
,
Luo Rongcheng
,
Han Huanxing
,
You Changxuan
,
Ding Xuemei
,
Li Aimin
,
Wang Chuanbin
,
Zhang Mingjang
2000, 7(4):288-290.
DOI: 10.3872/j.issn.1007-385X.2000.4.014
Abstract:
Objective: To evaluate the targeting activity in the animal model with human hepatoma, the 131 I human anti HBsAg Fab radioimmunoimaging was explored. Methods: Radioimmunoimagings were taken on different intervals after injection of 131 I human anti HBsAg Fab to the nude mice and tissue distribution was measured. The human anti HBsAg Fab was compared with the murine monoclonal antibodies. Results: The experimental group developed tumor positive images after 3 days of radio labeled monoclonal antibodies injection, and the peak accumulation of radio activity on the 5th day. Statistics indicated the tumor/liver ratio of the human anti HBsAg Fab, murine monoclonal antibodies and the control groups were 5.4,4.0 and 0.9 respectively on the 7th day. Conclusions: Our results suggest that the 131 I human anti HBsAg Fab has a considerable targeting activity, and provide an evidence that it can be used as a novel humanized carrer for targeting therapy of hepatoma.
Objective: To evaluate the targeting activity in the animal model with human hepatoma, the 131 I human anti HBsAg Fab radioimmunoimaging was explored. Methods: Radioimmunoimagings were taken on different intervals after injection of 131 I human anti HBsAg Fab to the nude mice and tissue distribution was measured. The human anti HBsAg Fab was compared with the murine monoclonal antibodies. Results: The experimental group developed tumor positive images after 3 days of radio labeled monoclonal antibodies injection, and the peak accumulation of radio activity on the 5th day. Statistics indicated the tumor/liver ratio of the human anti HBsAg Fab, murine monoclonal antibodies and the control groups were 5.4,4.0 and 0.9 respectively on the 7th day. Conclusions: Our results suggest that the 131 I human anti HBsAg Fab has a considerable targeting activity, and provide an evidence that it can be used as a novel humanized carrer for targeting therapy of hepatoma.
2000, 7(4):291-293.
DOI: 10.3872/j.issn.1007-385X.2000.4.015
Abstract:
Objective: To investigate the antitumor activities and apoptosis mechanisms of 3,7-dinitrodibenzobromonium salt (cBr) on human chronic myelogenous leukemia K562 cell line in vitro. Methods: The antitumor activities of cBr were tested by Trypanblau exclusion test ,MTT coloretric assay and clonal forming test,and the mechanisms were studied by morphological analysis(microscopy,fluorescencemicroscopy,electron microscopy)and flow cytometry analysis.Results: cBr could inhibit of the proliferation K562 cells and kill them significantly.The effects varied with different time and doses,and IC 50 of cell colony forming rate was 0.497 mmol/L.Apoptosis rate tested by flow cytometry was 70.34%,and apoptotic characteristics could be found by microscopy and electron microscopy. Conclusions: cBr could inhibit K562 cell proliferation greatly,and could induce K562 cells into apoptosis.
Objective: To investigate the antitumor activities and apoptosis mechanisms of 3,7-dinitrodibenzobromonium salt (cBr) on human chronic myelogenous leukemia K562 cell line in vitro. Methods: The antitumor activities of cBr were tested by Trypanblau exclusion test ,MTT coloretric assay and clonal forming test,and the mechanisms were studied by morphological analysis(microscopy,fluorescencemicroscopy,electron microscopy)and flow cytometry analysis.Results: cBr could inhibit of the proliferation K562 cells and kill them significantly.The effects varied with different time and doses,and IC 50 of cell colony forming rate was 0.497 mmol/L.Apoptosis rate tested by flow cytometry was 70.34%,and apoptotic characteristics could be found by microscopy and electron microscopy. Conclusions: cBr could inhibit K562 cell proliferation greatly,and could induce K562 cells into apoptosis.
2000, 7(4):296-297.
DOI: 10.3872/j.issn.1007-385X.2000.4.017
Abstract:
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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
- 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
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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.