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Volume 8,Issue 2,2001 Table of Contents
2001, 8(2):77-79.
DOI: 10.3872/j.issn.1007-385X.2001.2.001
Abstract:
SHI Ming
,
WANG Fu sheng
,
LIU Ming xu
,
ZHANG Bing
,
LEI Zhou yun
,
QIU Zhao hua
,
GAO Lan xing
,
WU Zu ze
2001, 8(2):80-83.
DOI: 10.3872/j.issn.1007-385X.2001.2.002
Abstract:
观察表达人野生型p5 3 ,GM CSF和B7 1基因的重组腺病毒载体 (BB 10 2 )提高肝癌细胞对化疗药物的敏感性。方法 :BEL 74 0 2 ,HLE和HuH 73株肝癌细胞被 5 0MOI的BB 10 2转染后 ,Westernblot检测p5 3基因的表达 ,MTT方法检测肝癌细胞对化疗药物的敏感性。结果 :当MOI为 5 0pfu/细胞时 ,腺病毒载体对 3株肝癌细胞的转染效率均达到 80 %以上。转染BB 10 2后 ,HLE和HuH 7细胞对顺铂的敏感性分别提高 11倍和 2 8倍 ,对丝裂霉素 C的敏感性分别提高 3 .75倍和 2 0倍。而BB 10 2的转染对BEL 74 0 2细胞的化疗敏感性没有影响。结论 :BB 10 2腺病毒载体转染后能显著提高p5 3基因突变的HLE和HuH 7细胞对顺铂和丝裂霉素 C的敏感性 ,因而增强顺铂和丝裂霉素 C对HLE和HuH 7细胞的杀伤作用
观察表达人野生型p5 3 ,GM CSF和B7 1基因的重组腺病毒载体 (BB 10 2 )提高肝癌细胞对化疗药物的敏感性。方法 :BEL 74 0 2 ,HLE和HuH 73株肝癌细胞被 5 0MOI的BB 10 2转染后 ,Westernblot检测p5 3基因的表达 ,MTT方法检测肝癌细胞对化疗药物的敏感性。结果 :当MOI为 5 0pfu/细胞时 ,腺病毒载体对 3株肝癌细胞的转染效率均达到 80 %以上。转染BB 10 2后 ,HLE和HuH 7细胞对顺铂的敏感性分别提高 11倍和 2 8倍 ,对丝裂霉素 C的敏感性分别提高 3 .75倍和 2 0倍。而BB 10 2的转染对BEL 74 0 2细胞的化疗敏感性没有影响。结论 :BB 10 2腺病毒载体转染后能显著提高p5 3基因突变的HLE和HuH 7细胞对顺铂和丝裂霉素 C的敏感性 ,因而增强顺铂和丝裂霉素 C对HLE和HuH 7细胞的杀伤作用
2001, 8(2):84-87.
DOI: 10.3872/j.issn.1007-385X.2001.2.003
Abstract:
利用bcl xS基因降低肿瘤细胞凋亡阈值 ,促进化疗药物顺铂诱导的鼻咽癌细胞凋亡 ,探索提高顺铂疗效的新途径。方法 :利用脂质体LipofectAmine将含有人bcl xS基因的重组真核表达质粒pcDNA3xS导入人鼻咽癌低分化上皮细胞株CNE 2Z ,G4 18筛选培养后 ,经Westernblot检测bcl xS的表达。以不含有bcl xS基因的pcDNA3质粒转染CNE 2Z作为对照细胞(CNE 2Zneo)。顺铂处理 2种细胞后 ,台盼蓝计数法求得各自的IC50 ,流式细胞仪及荧光染色检测凋亡细胞百分比。结果 :Westernblot检测证实获得稳定表达bcl xS的细胞 (CNE 2ZxS) ,与对照细胞相比 ,其对顺铂的IC50 值明显降低 ,经相同剂量顺铂处理后 ,流式细胞仪及荧光染色检测 ,结果表明 ,其凋亡细胞百分比明显高于对照细胞。结论 :bcl xS能显著增加鼻咽癌CNE 2Z细胞对顺铂诱导凋亡的敏感性
利用bcl xS基因降低肿瘤细胞凋亡阈值 ,促进化疗药物顺铂诱导的鼻咽癌细胞凋亡 ,探索提高顺铂疗效的新途径。方法 :利用脂质体LipofectAmine将含有人bcl xS基因的重组真核表达质粒pcDNA3xS导入人鼻咽癌低分化上皮细胞株CNE 2Z ,G4 18筛选培养后 ,经Westernblot检测bcl xS的表达。以不含有bcl xS基因的pcDNA3质粒转染CNE 2Z作为对照细胞(CNE 2Zneo)。顺铂处理 2种细胞后 ,台盼蓝计数法求得各自的IC50 ,流式细胞仪及荧光染色检测凋亡细胞百分比。结果 :Westernblot检测证实获得稳定表达bcl xS的细胞 (CNE 2ZxS) ,与对照细胞相比 ,其对顺铂的IC50 值明显降低 ,经相同剂量顺铂处理后 ,流式细胞仪及荧光染色检测 ,结果表明 ,其凋亡细胞百分比明显高于对照细胞。结论 :bcl xS能显著增加鼻咽癌CNE 2Z细胞对顺铂诱导凋亡的敏感性
2001, 8(2):93-97.
DOI: 10.3872/j.issn.1007-385X.2001.2.005
Abstract:
探讨VP2 2的细胞间传递作用对HSV tk/GCV杀伤肿瘤细胞的促进作用。方法 :将VP2 2与报告基因 绿色荧光蛋白基因 (GFP)或前药酶基因 单纯疱疹病毒胸苷激酶基因 (HSV tk)构建在同一载体上 ,进行DNA序列分析鉴定 ;将此融合基因载体转染 2 93T或COS7细胞 ,免疫荧光分析确定转染情况和细胞间传递作用 ;在前药GCV不同浓度、转染与未转染细胞比例不同时 ,MTT法检测细胞增殖情况 ;利用转染细胞培养液上清培养未转染细胞 ,确定旁观者效应是否由培养液转移。结果 :PCR及序列分析显示融合基因插入载体正确 ;对 2 93T或COS7细胞转染率可达 2 5 %～ 3 0 % ,且证明融合蛋白已被表达并在细胞间传递 ;HSV tk与VP2 2融合后在前药GCV浓度低至 0 .1μg/ml时就可显示出细胞杀伤效应增强 ,并随着表达VP2 2 HSV tk细胞比例的增加 ,细胞杀伤作用增强。结论 :VP2 2介导的自杀基因产物的细胞间传递作用可解决自杀基因的低效细胞毒作用 ,明显增强细胞杀伤效应。
探讨VP2 2的细胞间传递作用对HSV tk/GCV杀伤肿瘤细胞的促进作用。方法 :将VP2 2与报告基因 绿色荧光蛋白基因 (GFP)或前药酶基因 单纯疱疹病毒胸苷激酶基因 (HSV tk)构建在同一载体上 ,进行DNA序列分析鉴定 ;将此融合基因载体转染 2 93T或COS7细胞 ,免疫荧光分析确定转染情况和细胞间传递作用 ;在前药GCV不同浓度、转染与未转染细胞比例不同时 ,MTT法检测细胞增殖情况 ;利用转染细胞培养液上清培养未转染细胞 ,确定旁观者效应是否由培养液转移。结果 :PCR及序列分析显示融合基因插入载体正确 ;对 2 93T或COS7细胞转染率可达 2 5 %～ 3 0 % ,且证明融合蛋白已被表达并在细胞间传递 ;HSV tk与VP2 2融合后在前药GCV浓度低至 0 .1μg/ml时就可显示出细胞杀伤效应增强 ,并随着表达VP2 2 HSV tk细胞比例的增加 ,细胞杀伤作用增强。结论 :VP2 2介导的自杀基因产物的细胞间传递作用可解决自杀基因的低效细胞毒作用 ,明显增强细胞杀伤效应。
2001, 8(2):98-101.
DOI: 10.3872/j.issn.1007-385X.2001.2.006
Abstract:
探讨趋化因子MCP 3能否增强B7基因诱导小鼠抗宫颈癌主动免疫的效果。方法 :实验组 :将pCMV/MCP 3以基因药物的方式肌注 615小鼠 ,同时用B7+U14细胞疫苗在同一部位免疫小鼠 ,再将野生型U14移植入已免疫的小鼠。对照组A :用B7+U14细胞疫苗 +生理盐水同时免疫 615系小鼠 ;对照组B :用pCMV/MCP 3 +野生型U14免疫小鼠 ;其它处理同治疗组。观察 3组小鼠的成瘤情况、生存期。用以上 3种方案分别处理小鼠 ,2周后分别取脾T淋巴细胞用MTT法体外检测其对U14的杀伤率。结果 :经方差分析比较 3组在移植U14后不同时程瘤体平均体积有显著性差异 (F =91.86,P <0 .0 0 1)。3组平均生存期分别为 10 0 ,63 ,3 7d ,经Kaplan Meier曲线比较 ,差异具显著性 (P <0 .0 1)。体外检测经MCP 3和B7+U14疫苗处理的小鼠 ,其T淋巴细胞在不同效靶比对U14的杀伤效率均明显高于经MCP 3和U14疫苗处理者 (F =4 0 62 ,P <0 .0 0 1)。结论 :趋化因子MPC 3能增强B7基因修饰的U14细胞疫苗的抗肿瘤效果 ,其增强作用需疫苗有B7的表达为前提
探讨趋化因子MCP 3能否增强B7基因诱导小鼠抗宫颈癌主动免疫的效果。方法 :实验组 :将pCMV/MCP 3以基因药物的方式肌注 615小鼠 ,同时用B7+U14细胞疫苗在同一部位免疫小鼠 ,再将野生型U14移植入已免疫的小鼠。对照组A :用B7+U14细胞疫苗 +生理盐水同时免疫 615系小鼠 ;对照组B :用pCMV/MCP 3 +野生型U14免疫小鼠 ;其它处理同治疗组。观察 3组小鼠的成瘤情况、生存期。用以上 3种方案分别处理小鼠 ,2周后分别取脾T淋巴细胞用MTT法体外检测其对U14的杀伤率。结果 :经方差分析比较 3组在移植U14后不同时程瘤体平均体积有显著性差异 (F =91.86,P <0 .0 0 1)。3组平均生存期分别为 10 0 ,63 ,3 7d ,经Kaplan Meier曲线比较 ,差异具显著性 (P <0 .0 1)。体外检测经MCP 3和B7+U14疫苗处理的小鼠 ,其T淋巴细胞在不同效靶比对U14的杀伤效率均明显高于经MCP 3和U14疫苗处理者 (F =4 0 62 ,P <0 .0 0 1)。结论 :趋化因子MPC 3能增强B7基因修饰的U14细胞疫苗的抗肿瘤效果 ,其增强作用需疫苗有B7的表达为前提
2001, 8(2):102-105.
DOI: 10.3872/j.issn.1007-385X.2001.2.007
Abstract:
探讨溶血链球菌冻干制剂OK 4 3 2的抗肿瘤机理。方法 :B16黑色素瘤细胞接种于C5 7BL/ 6小鼠皮下 ,3d后腹腔注射 1KU的OK 4 3 2 ,每周 1次 ,连续 3周。观察肿瘤生长体积及荷瘤小鼠的生存期 ,并测定了OK 4 3 2在体内、体外对荷瘤小鼠脾细胞IL 2 ,IL 4 ,IL 6,IL 10 ,IL 12 ,IFN γ分泌的影响。结果 :OK 4 3 2能显著抑制B16黑色素瘤的生长 ,延长荷瘤小鼠的生存期 (P <0 .0 5 ) ;在体外OK 4 3 2可刺激荷瘤小鼠脾细胞IL 6,IL 10 ,IL 12 ,IFN γ的分泌 (P <0 .0 1) ;腹腔注射OK 4 3 2后荷瘤小鼠脾细胞IL 2 ,IL 12 ,IFN γ的分泌显著增加 ,而IL 10显著减少 (P <0 .0 5 ) ,提示OK 4 3 2治疗后荷瘤小鼠脾细胞Th1增加 ,Th2 相对减少。结论 :OK 4 3 2在体内可通过诱导荷瘤小鼠脾细胞IL 12的分泌 ,促进Th0向Th1分化 ,使宿主的免疫功能处于Th1优势状态 ,从而增强宿主的抗肿瘤作用
探讨溶血链球菌冻干制剂OK 4 3 2的抗肿瘤机理。方法 :B16黑色素瘤细胞接种于C5 7BL/ 6小鼠皮下 ,3d后腹腔注射 1KU的OK 4 3 2 ,每周 1次 ,连续 3周。观察肿瘤生长体积及荷瘤小鼠的生存期 ,并测定了OK 4 3 2在体内、体外对荷瘤小鼠脾细胞IL 2 ,IL 4 ,IL 6,IL 10 ,IL 12 ,IFN γ分泌的影响。结果 :OK 4 3 2能显著抑制B16黑色素瘤的生长 ,延长荷瘤小鼠的生存期 (P <0 .0 5 ) ;在体外OK 4 3 2可刺激荷瘤小鼠脾细胞IL 6,IL 10 ,IL 12 ,IFN γ的分泌 (P <0 .0 1) ;腹腔注射OK 4 3 2后荷瘤小鼠脾细胞IL 2 ,IL 12 ,IFN γ的分泌显著增加 ,而IL 10显著减少 (P <0 .0 5 ) ,提示OK 4 3 2治疗后荷瘤小鼠脾细胞Th1增加 ,Th2 相对减少。结论 :OK 4 3 2在体内可通过诱导荷瘤小鼠脾细胞IL 12的分泌 ,促进Th0向Th1分化 ,使宿主的免疫功能处于Th1优势状态 ,从而增强宿主的抗肿瘤作用
2001, 8(2):106-109.
DOI: 10.3872/j.issn.1007-385X.2001.2.008
Abstract:
研究不同浓度As2 O3对食管癌细胞系的作用。方法 :食管恶性转化上皮细胞 (SHEEC1)是我室用人乳头状瘤病毒 (HPV) 18和TPA诱发的细胞系 ,培养在培养瓶和 2 4孔板 ,用浓度 1,3和 5 μmol/L的As2 O3处理 2 ,4 ,8,16,2 4h ,电镜检查细胞形态 ,光镜检查核分裂指数 (MI) ;用放射自显影检查 [3H] TdR掺入细胞 ;流式细胞仪检查增殖指数 (PI)和凋亡指数 (AI)。结果 :低剂量As2 O3( 1.0 μmol/L)组 [3H] TdR掺入细胞 ,MI,PI皆增加 ,提示SHEEC1增殖率提高 ;高剂量组 ( 3和 5 μmol/L)高AI和低PI,给药 2 4h后 ,电镜可见细胞凋亡和坏死。结论 :不同剂量氧化砷的作用不同 ,低剂量可引起食管癌细胞DNA合成增加和促进细胞增殖 ,高剂量则引起细胞凋亡和坏死。
研究不同浓度As2 O3对食管癌细胞系的作用。方法 :食管恶性转化上皮细胞 (SHEEC1)是我室用人乳头状瘤病毒 (HPV) 18和TPA诱发的细胞系 ,培养在培养瓶和 2 4孔板 ,用浓度 1,3和 5 μmol/L的As2 O3处理 2 ,4 ,8,16,2 4h ,电镜检查细胞形态 ,光镜检查核分裂指数 (MI) ;用放射自显影检查 [3H] TdR掺入细胞 ;流式细胞仪检查增殖指数 (PI)和凋亡指数 (AI)。结果 :低剂量As2 O3( 1.0 μmol/L)组 [3H] TdR掺入细胞 ,MI,PI皆增加 ,提示SHEEC1增殖率提高 ;高剂量组 ( 3和 5 μmol/L)高AI和低PI,给药 2 4h后 ,电镜可见细胞凋亡和坏死。结论 :不同剂量氧化砷的作用不同 ,低剂量可引起食管癌细胞DNA合成增加和促进细胞增殖 ,高剂量则引起细胞凋亡和坏死。
2001, 8(2):110-113.
DOI: 10.3872/j.issn.1007-385X.2001.2.009
Abstract:
Objective: To probe into the physical and chemical property, biology activity,acute and chronic toxicity of low molecular weight natural tumor suppressor (LMW NTS)of fetal bovine hepatocytes. Methods: LMW NTS was isolated from fetal bovine hepatocytes and its molecular weight was measured by high pressure liquid chromatography, the ratio of OD 260 /OD 280 was calculated by spectrophotomctry. Anti tumor effects of fetal bovine hepatocytes LMW NTS were investigated by in vitro two layers agar plate cell culture and by tumor bearing mice following the intraperitoneal administration of the fetal bovine hepatocytes LMW NTS. Results: The fetal bovine hepatocytes LMW NTS had approximate molecular weight of about 7 000~8 000 and belonged to low molecular weight ribonucleic acid and peptide. Under the condition of in vitro agar plate culture, the LMW NTS of fetal bovine hepatocytes showed remarkable inhibitory effects on the growth of murine S 180 cells, but weaker effects on the growth of normal murine bone marrow granulocyte macrophage progenitors. Survival period of the S 180 cells bearing mice treated with intraperitoneal injection of the fetal bovine hepatocytes LMW NTS was strikingly prolonged( P <0.001). Toxicity test proved that the fetal bovine hepatocytes LMW NTS had no side effects and was safe for long time application. Conclusion: The LMW NTS of fetal bovine hepatocytes possesses the property of low molecular weight and striking anti tumor activities both in vitro and in vivo.
Objective: To probe into the physical and chemical property, biology activity,acute and chronic toxicity of low molecular weight natural tumor suppressor (LMW NTS)of fetal bovine hepatocytes. Methods: LMW NTS was isolated from fetal bovine hepatocytes and its molecular weight was measured by high pressure liquid chromatography, the ratio of OD 260 /OD 280 was calculated by spectrophotomctry. Anti tumor effects of fetal bovine hepatocytes LMW NTS were investigated by in vitro two layers agar plate cell culture and by tumor bearing mice following the intraperitoneal administration of the fetal bovine hepatocytes LMW NTS. Results: The fetal bovine hepatocytes LMW NTS had approximate molecular weight of about 7 000~8 000 and belonged to low molecular weight ribonucleic acid and peptide. Under the condition of in vitro agar plate culture, the LMW NTS of fetal bovine hepatocytes showed remarkable inhibitory effects on the growth of murine S 180 cells, but weaker effects on the growth of normal murine bone marrow granulocyte macrophage progenitors. Survival period of the S 180 cells bearing mice treated with intraperitoneal injection of the fetal bovine hepatocytes LMW NTS was strikingly prolonged( P <0.001). Toxicity test proved that the fetal bovine hepatocytes LMW NTS had no side effects and was safe for long time application. Conclusion: The LMW NTS of fetal bovine hepatocytes possesses the property of low molecular weight and striking anti tumor activities both in vitro and in vivo.
2001, 8(2):114-117.
DOI: 10.3872/j.issn.1007-385X.2001.2.010
Abstract:
Objective: To investigate the mode of cell death caused by NDV strain Changchun and NDV strain Siping. Methods: Plaque formation, cell suppression test, gel electrophoresis, and TUNEL assay were used after the cells were infected by NDV. Results: The apparently pathological changes were observed in chicken embryo fibroblasts, BHK, Hela, Hep 2, HCT and OS 732 tumor cells, but not in Wish cells. The higher suppressed effect on tumor cells was found in the NDV strain Changchun than that in the NDV strain Siping. There was no dose effect relationship between NDV and tumor cell suppression, only optimum dose NDV could cause maximal tumor cells inhibitory effect. Conclusion: The mode of cell death might be different after infection of NDV. The NDV strain Changchun killed tumor cells mainly through apoptosis, while the NDV strain Siping killed tumor cells mainly through necrosis.
Objective: To investigate the mode of cell death caused by NDV strain Changchun and NDV strain Siping. Methods: Plaque formation, cell suppression test, gel electrophoresis, and TUNEL assay were used after the cells were infected by NDV. Results: The apparently pathological changes were observed in chicken embryo fibroblasts, BHK, Hela, Hep 2, HCT and OS 732 tumor cells, but not in Wish cells. The higher suppressed effect on tumor cells was found in the NDV strain Changchun than that in the NDV strain Siping. There was no dose effect relationship between NDV and tumor cell suppression, only optimum dose NDV could cause maximal tumor cells inhibitory effect. Conclusion: The mode of cell death might be different after infection of NDV. The NDV strain Changchun killed tumor cells mainly through apoptosis, while the NDV strain Siping killed tumor cells mainly through necrosis.
2001, 8(2):118-121.
DOI: 10.3872/j.issn.1007-385X.2001.2.011
Abstract:
Objective: To explore the charateristics of human hepatocarcinoma cells growing in various strains of mice with different immunity and T lymphocyte rejection to the hepatocarcinoma cells. Methods: The human hepatocarcinoma cells were inoculated into various strains of mice with different immunity (including BALB/c, CBA/N, BALB/c nu, SCID and BALB/c PBL SCID,CBA/N PBL SCID) and their growths were surveyed. Then the spleen cells were harvested to examine their direct cytotoxicity to the hepatocarcinoma cells.The percentage of CD4 and CD8 cell of peripheral blood from mice was assyed by FCM. Results: There was no carcinoma growth in BALB/c and BALB/c PBL SCID,but was 100% carcinoma growth in SICD, nude and CBA/N PBL SCID. The spleen cells in experimental groups from BALB/c and CBA/N mice displayed strong cytotoxicity to target cells and those of immunoreconstituted SCID generated a little lysis to target cells, but those of control group and nude, SCID mice generated no lysis to target cells.The percentage of CD4 from BALB/c and CBA/N mice in experimental groups decreased,but the percentage of CD8 changed not much while CD4 /CD8 ratio decreased.Couclusion: Whether the hepatocarcinoma cells grew in mice correlated strongly with T lymphocytes which were xenospecific cytotoxicity and played a main role of immune killing in rejection to xenografted tumor.
Objective: To explore the charateristics of human hepatocarcinoma cells growing in various strains of mice with different immunity and T lymphocyte rejection to the hepatocarcinoma cells. Methods: The human hepatocarcinoma cells were inoculated into various strains of mice with different immunity (including BALB/c, CBA/N, BALB/c nu, SCID and BALB/c PBL SCID,CBA/N PBL SCID) and their growths were surveyed. Then the spleen cells were harvested to examine their direct cytotoxicity to the hepatocarcinoma cells.The percentage of CD4 and CD8 cell of peripheral blood from mice was assyed by FCM. Results: There was no carcinoma growth in BALB/c and BALB/c PBL SCID,but was 100% carcinoma growth in SICD, nude and CBA/N PBL SCID. The spleen cells in experimental groups from BALB/c and CBA/N mice displayed strong cytotoxicity to target cells and those of immunoreconstituted SCID generated a little lysis to target cells, but those of control group and nude, SCID mice generated no lysis to target cells.The percentage of CD4 from BALB/c and CBA/N mice in experimental groups decreased,but the percentage of CD8 changed not much while CD4 /CD8 ratio decreased.Couclusion: Whether the hepatocarcinoma cells grew in mice correlated strongly with T lymphocytes which were xenospecific cytotoxicity and played a main role of immune killing in rejection to xenografted tumor.
2001, 8(2):122-125.
DOI: 10.3872/j.issn.1007-385X.2001.2.012
Abstract:
Objective: To investigate the role and mechanism of histamine in regulation of C57BL/6 mice spleen derived LAK activity in vivo. Methods: The C57BL/6 mice carrying B16 pulmonary metastatic melanoma were treated with LAK/IL 2/histamine therapy or LAK/IL 2 therapy with the aim of evaluating both anti tumor responses in vivo. Results: It was found that the addition of histamine effectively potentiated the anti metastatic effect produced by LAK/IL 2 therapy and induced regression of NK resistant B16 pulmonary metastatic melanoma. Survival period was significantly prolonged in mice receiving LAK/IL 2/histamine as compared with LAK/IL 2 therapy alone. The effect of histamine was completely blocked by H 2 R antagonist ranitidine, and mimicked by dimaprit, a H 2 R agonist. Conclusion:Histamine, via specific activation of H 2 R, may be an important regulator of LAK cells activity; histamine and LAK/IL 2 synergistically induce more potent antitumor efficacy in vivo .
Objective: To investigate the role and mechanism of histamine in regulation of C57BL/6 mice spleen derived LAK activity in vivo. Methods: The C57BL/6 mice carrying B16 pulmonary metastatic melanoma were treated with LAK/IL 2/histamine therapy or LAK/IL 2 therapy with the aim of evaluating both anti tumor responses in vivo. Results: It was found that the addition of histamine effectively potentiated the anti metastatic effect produced by LAK/IL 2 therapy and induced regression of NK resistant B16 pulmonary metastatic melanoma. Survival period was significantly prolonged in mice receiving LAK/IL 2/histamine as compared with LAK/IL 2 therapy alone. The effect of histamine was completely blocked by H 2 R antagonist ranitidine, and mimicked by dimaprit, a H 2 R agonist. Conclusion:Histamine, via specific activation of H 2 R, may be an important regulator of LAK cells activity; histamine and LAK/IL 2 synergistically induce more potent antitumor efficacy in vivo .
2001, 8(2):126-129.
DOI: 10.3872/j.issn.1007-385X.2001.2.013
Abstract:
探讨榄香烯复合瘤苗 (EC TCV)对小鼠肝癌Hca F的免疫治疗效应及治疗小鼠脾细胞IL 10与IL 12分泌的变化。方法 :带Hca F小鼠分别用EC TCV ,CY ,EC TCV +CY治疗或不治疗 (PBS对照 ) ,观察比较肿瘤生长情况并用ELISA法测脾细胞与Hca F细胞混合培养上清中的IL 10与IL 12。结果 :EC TCV免疫治疗或CY化疗后 ,肿瘤潜伏期延长 ,瘤重减轻 ,用EC TCV +CY进行化免治疗的小鼠在第 15天活杀时均无瘤生长 ,与Hca F细胞混合培养的EC TCV治疗小鼠脾细胞培养上清中IL 10浓度明显降低 ( 1.82± 0 .2 9pg/ml,P <0 .0 5 ) ,IL 12浓度明显升高 ( 7.13± 1.89pg/ml,P <0 .0 5 )。结论 :EC TCV对Hca F有免疫治疗效应 ,可与CY并用而加强 ,而EC TCV主动免疫促进脾细胞IL 12分泌、抑制IL 10分泌可能是其抗瘤作用的机制之一。
探讨榄香烯复合瘤苗 (EC TCV)对小鼠肝癌Hca F的免疫治疗效应及治疗小鼠脾细胞IL 10与IL 12分泌的变化。方法 :带Hca F小鼠分别用EC TCV ,CY ,EC TCV +CY治疗或不治疗 (PBS对照 ) ,观察比较肿瘤生长情况并用ELISA法测脾细胞与Hca F细胞混合培养上清中的IL 10与IL 12。结果 :EC TCV免疫治疗或CY化疗后 ,肿瘤潜伏期延长 ,瘤重减轻 ,用EC TCV +CY进行化免治疗的小鼠在第 15天活杀时均无瘤生长 ,与Hca F细胞混合培养的EC TCV治疗小鼠脾细胞培养上清中IL 10浓度明显降低 ( 1.82± 0 .2 9pg/ml,P <0 .0 5 ) ,IL 12浓度明显升高 ( 7.13± 1.89pg/ml,P <0 .0 5 )。结论 :EC TCV对Hca F有免疫治疗效应 ,可与CY并用而加强 ,而EC TCV主动免疫促进脾细胞IL 12分泌、抑制IL 10分泌可能是其抗瘤作用的机制之一。
CHEN Guo you
,
JIANG Ying ming
,
Zhang Yi
,
HUANG Xin
,
LI Yong jian
,
SHI Qun ying
,
WANG Quan xing
,
ZHANG Min hui
,
HE Long
,
CAO Xue tao
2001, 8(2):130-133.
DOI: 10.3872/j.issn.1007-385X.2001.2.014
Abstract:
摸索一种适合于中试生产的点突变重组人β干扰素(17Ser-IFN-β)发酵与纯化工艺。方法研究不同培养基和热诱导时间对17Ser-IFN-β表达的影响;观察pH、蛋白浓度和层析条件对17Ser-IFN-β纯化的影响。结果17Ser-IFN-β工程菌在含甘油、酪蛋白水解物及微量元素的M9培养基中，17Ser-IFN-β表达量明显增加。灰度扫描显示热诱导4h后的表达量最高，目的蛋白可达菌体总蛋白的20%以上。17Ser-IFN-β可以相对特异性地被仲丁醇抽提至有机相中，纯度达80%，经S200凝胶过滤柱层析后纯度达90%，用G25柱转换缓冲液，使缓冲液pH值＞8，同时去除缓冲液中的还原剂DTT，将复性后的样品用C4反相柱层析进行精制，最终获得纯度＞98%和比活性>3×107U/mg的17Ser-IFN-β制品。结论成功地建立了注射用新型重组人β-干扰素的中试生产工艺，为随后进行的临床前研究及临床试验打下基础。
摸索一种适合于中试生产的点突变重组人β干扰素(17Ser-IFN-β)发酵与纯化工艺。方法研究不同培养基和热诱导时间对17Ser-IFN-β表达的影响;观察pH、蛋白浓度和层析条件对17Ser-IFN-β纯化的影响。结果17Ser-IFN-β工程菌在含甘油、酪蛋白水解物及微量元素的M9培养基中，17Ser-IFN-β表达量明显增加。灰度扫描显示热诱导4h后的表达量最高，目的蛋白可达菌体总蛋白的20%以上。17Ser-IFN-β可以相对特异性地被仲丁醇抽提至有机相中，纯度达80%，经S200凝胶过滤柱层析后纯度达90%，用G25柱转换缓冲液，使缓冲液pH值＞8，同时去除缓冲液中的还原剂DTT，将复性后的样品用C4反相柱层析进行精制，最终获得纯度＞98%和比活性>3×107U/mg的17Ser-IFN-β制品。结论成功地建立了注射用新型重组人β-干扰素的中试生产工艺，为随后进行的临床前研究及临床试验打下基础。
SONG Wen gang
,
YU Yi zhi
,
QU Xun
,
LIU Shu xun
,
WANG Wen ya
,
ZHANG Ming hui
,
TANG Ling
,
LI Yong jian
,
CAO Xue tao
2001, 8(2):134-137.
DOI: 10.3872/j.issn.1007-385X.2001.2.015
Abstract:
探讨肿瘤细胞对树突状细胞 (dendriticcells ,DC)分化发育的影响。方法 :在诱导小鼠骨髓DC发育的过程中 ,加入肿瘤细胞培养上清 ,观察DC生成情况 ,并与正常培养条件下的DC比较共刺激分子、Ⅱ类分子、黏附分子的表达 ,刺激同种T细胞增殖 ,以及抗原提呈功能水平。通过ELISA法检测肿瘤细胞分泌的细胞因子。结果 :经小鼠黑色素细胞瘤B16和肺癌细胞 3LL的分泌上清培养后 ,DC表达共刺激分子CD4 0 ,CD86,CD80 ,和黏附分子CD5 4的水平下降 ;分泌Th1细胞因子IL 12的水平也降低 ;并且DC的抗原提呈功能和刺激同种混合淋巴细胞反应的功能下降。上述肿瘤细胞均可分泌抑制性细胞因子IL 10。结论 :在肿瘤组织的微环境中存在免疫抑制性因子 ,对DC分化发育和抗原提呈功能起负性调节作用
探讨肿瘤细胞对树突状细胞 (dendriticcells ,DC)分化发育的影响。方法 :在诱导小鼠骨髓DC发育的过程中 ,加入肿瘤细胞培养上清 ,观察DC生成情况 ,并与正常培养条件下的DC比较共刺激分子、Ⅱ类分子、黏附分子的表达 ,刺激同种T细胞增殖 ,以及抗原提呈功能水平。通过ELISA法检测肿瘤细胞分泌的细胞因子。结果 :经小鼠黑色素细胞瘤B16和肺癌细胞 3LL的分泌上清培养后 ,DC表达共刺激分子CD4 0 ,CD86,CD80 ,和黏附分子CD5 4的水平下降 ;分泌Th1细胞因子IL 12的水平也降低 ;并且DC的抗原提呈功能和刺激同种混合淋巴细胞反应的功能下降。上述肿瘤细胞均可分泌抑制性细胞因子IL 10。结论 :在肿瘤组织的微环境中存在免疫抑制性因子 ,对DC分化发育和抗原提呈功能起负性调节作用
2001, 8(2):138-141.
DOI: 10.3872/j.issn.1007-385X.2001.2.016
Abstract:
构建一个鼠源性的抗内毒素单链噬菌体抗体库，从中筛选出对内毒素具有较高亲和力的单链抗体。方法从小鼠脾细胞中提取总RNA，通过RT-PCR技术扩增出小鼠抗体重链、轻链可变区基因（VH，VL），用Linker将VH，VL交联形成单链抗体可变区片段（ScFv）。经NotⅠ，SfiⅠ双酶切后与经同样双酶切的pCANTAB5E载体相连，转化入大肠杆菌TG1以构建鼠抗内毒素单链噬菌体抗体库。在援救噬菌体抗体库后，用内毒素淘筛特异性的ScFv，富集的噬菌体阳性克隆重新感染TG1。在96孔板分别援救单个含特异性ScFv的TG1菌落，最后随机挑选出190个菌落经ELISA检测抗内毒素ScFv。结果小鼠血清中抗内毒素的效价为1∶12800。提取的总RNA浓度为12.3813μg/ml，纯度较好。扩增出的VH长约340bp，VL约320bp，ScFv约800bp。转化入TG1后有约1.9×107个菌落。淘筛一轮过后即有3×104阳性菌落长出，190个菌落经ELISA检测有2个阳性克隆。结论成功地构建了一个库容量为1.9×107的鼠抗内毒素单链噬菌体抗体库，并从中筛选出了2株抗内毒素ScFv。
构建一个鼠源性的抗内毒素单链噬菌体抗体库，从中筛选出对内毒素具有较高亲和力的单链抗体。方法从小鼠脾细胞中提取总RNA，通过RT-PCR技术扩增出小鼠抗体重链、轻链可变区基因（VH，VL），用Linker将VH，VL交联形成单链抗体可变区片段（ScFv）。经NotⅠ，SfiⅠ双酶切后与经同样双酶切的pCANTAB5E载体相连，转化入大肠杆菌TG1以构建鼠抗内毒素单链噬菌体抗体库。在援救噬菌体抗体库后，用内毒素淘筛特异性的ScFv，富集的噬菌体阳性克隆重新感染TG1。在96孔板分别援救单个含特异性ScFv的TG1菌落，最后随机挑选出190个菌落经ELISA检测抗内毒素ScFv。结果小鼠血清中抗内毒素的效价为1∶12800。提取的总RNA浓度为12.3813μg/ml，纯度较好。扩增出的VH长约340bp，VL约320bp，ScFv约800bp。转化入TG1后有约1.9×107个菌落。淘筛一轮过后即有3×104阳性菌落长出，190个菌落经ELISA检测有2个阳性克隆。结论成功地构建了一个库容量为1.9×107的鼠抗内毒素单链噬菌体抗体库，并从中筛选出了2株抗内毒素ScFv。
2001, 8(2):142-144.
DOI: 10.3872/j.issn.1007-385X.2001.2.017
Abstract:
Objective: To adjust the distance between SD sequence and ATG in the same expressive plasmid pLX1 to enhance expression of heterologous bFGF gene in E. coli. Methods: Adjusting the different distance between SD sequence and ATG by Klenow and Mung Bean Nuclease. SDS PAGE and Western blot showed the expressed protein bFGF in E.coli. bFGF proteins were purified by HPHIC, HPGFC and HAC. Biological activity was examined by MTT. Results: Recombinant plasmids pLX2, pLX3 were obtained and the expressive levels were 8.03%, 9.9% respectively. Also the purified bFGF was obtained by HPHIC, HPGFC, HAC and its ED 50 was 2.29 ng/ml. Conclusion: Increasing the bFGF gene dosage by adjusting the distance between SD sequence and ATG could increase the expression level of a desired protein.
Objective: To adjust the distance between SD sequence and ATG in the same expressive plasmid pLX1 to enhance expression of heterologous bFGF gene in E. coli. Methods: Adjusting the different distance between SD sequence and ATG by Klenow and Mung Bean Nuclease. SDS PAGE and Western blot showed the expressed protein bFGF in E.coli. bFGF proteins were purified by HPHIC, HPGFC and HAC. Biological activity was examined by MTT. Results: Recombinant plasmids pLX2, pLX3 were obtained and the expressive levels were 8.03%, 9.9% respectively. Also the purified bFGF was obtained by HPHIC, HPGFC, HAC and its ED 50 was 2.29 ng/ml. Conclusion: Increasing the bFGF gene dosage by adjusting the distance between SD sequence and ATG could increase the expression level of a desired protein.
2001, 8(2):153-154.
DOI: 10.3872/j.issn.1007-385X.2001.2.022
Abstract:
超抗原是一类只需极低浓度（1－10ng/ml)即可激活大量的淋巴细胞克隆，产生极强的应答效应的抗原性物质。葡萄球菌肠毒素A（staphylococcal enterotoxinA,SEA)是金黄色葡萄球菌产生的葡萄球菌肠毒素家族中一员，是最有效的T细胞活化因子之一。研究发现，SEA可通过刺激T细胞增殖，促进IL－2，TNF－α，IFN－γ等淋巴因子的产生，从而发挥抗肿瘤治疗作用。
超抗原是一类只需极低浓度（1－10ng/ml)即可激活大量的淋巴细胞克隆，产生极强的应答效应的抗原性物质。葡萄球菌肠毒素A（staphylococcal enterotoxinA,SEA)是金黄色葡萄球菌产生的葡萄球菌肠毒素家族中一员，是最有效的T细胞活化因子之一。研究发现，SEA可通过刺激T细胞增殖，促进IL－2，TNF－α，IFN－γ等淋巴因子的产生，从而发挥抗肿瘤治疗作用。
2001, 8(2):155-156.
DOI: 10.3872/j.issn.1007-385X.2001.2.023
Abstract:
自杀基因疗法是近几年来的研究热点 ,最初被应用于脑胶质瘤的临床治疗 ,以后又推广到其他恶性肿瘤的治疗。但是 ,这种疗法存在许多缺陷 ,其临床疗效并不令人满意。近年来有学者尝试利用辐射来实现对自杀基因体内表达的时空调控 ,提高基因靶向转移的效率 ,同时自杀基因疗法还可以提高肿瘤对放射治疗的敏感性。本文将对这方面的最新研究成果作一综述。
自杀基因疗法是近几年来的研究热点 ,最初被应用于脑胶质瘤的临床治疗 ,以后又推广到其他恶性肿瘤的治疗。但是 ,这种疗法存在许多缺陷 ,其临床疗效并不令人满意。近年来有学者尝试利用辐射来实现对自杀基因体内表达的时空调控 ,提高基因靶向转移的效率 ,同时自杀基因疗法还可以提高肿瘤对放射治疗的敏感性。本文将对这方面的最新研究成果作一综述。
2001, 8(2):157-159.
DOI: 10.3872/j.issn.1007-385X.2001.2.024
Abstract:
TRAIL是近年来发现的又一凋亡分子 ,它的只诱导肿瘤细胞凋亡而不对机体正常组织产生毒性效应的特征使其在肿瘤的免疫治疗中具有潜在的应用前景。然而将TRAIL应用于临床还需要对其作用机制、抗瘤效应、以及毒性等方面进行深入探索。本文介绍了TRAIL对肿瘤细胞的凋亡诱导的作用机制 ,和国外对TRAIL的体内外抗瘤实验观察 ,联合化疗等效应以及TRAIL可能存在的潜在毒性 ,为将来TRAIL的临床研究提供一定的线索
TRAIL是近年来发现的又一凋亡分子 ,它的只诱导肿瘤细胞凋亡而不对机体正常组织产生毒性效应的特征使其在肿瘤的免疫治疗中具有潜在的应用前景。然而将TRAIL应用于临床还需要对其作用机制、抗瘤效应、以及毒性等方面进行深入探索。本文介绍了TRAIL对肿瘤细胞的凋亡诱导的作用机制 ,和国外对TRAIL的体内外抗瘤实验观察 ,联合化疗等效应以及TRAIL可能存在的潜在毒性 ,为将来TRAIL的临床研究提供一定的线索
2001, 8(2):160-162.
DOI: 10.3872/j.issn.1007-385X.2001.2.025
Abstract:
本文介绍了靶向腺病毒与传统腺病毒载体相比之优势以及两种不同的靶向腺病毒构建策略，靶向腺病毒需要满足两个条件，一是去除天然嗜性，二是靶向新受体。有两种途径可以到这一目的，“双组分体系”中，腺病毒与双特异分子结合，该双特异分子可同时去除天然嗜性和靶向新受体。“单组分体系”中，病毒颗粒经基因修饰后，不再与天然受体结合，并加入了新配基。双组分体系可灵活地通过某特受体而达到靶向效果，单组分体系则更适于作用为基因治疗药物。
本文介绍了靶向腺病毒与传统腺病毒载体相比之优势以及两种不同的靶向腺病毒构建策略，靶向腺病毒需要满足两个条件，一是去除天然嗜性，二是靶向新受体。有两种途径可以到这一目的，“双组分体系”中，腺病毒与双特异分子结合，该双特异分子可同时去除天然嗜性和靶向新受体。“单组分体系”中，病毒颗粒经基因修饰后，不再与天然受体结合，并加入了新配基。双组分体系可灵活地通过某特受体而达到靶向效果，单组分体系则更适于作用为基因治疗药物。
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- 《中国肿瘤生物治疗杂志》
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- 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.