壳聚糖作为基因递送载体的MUC1基因疫苗治疗胰腺癌的实验研究

【摘要】 目的 研究以壳聚糖作为基因递送载体的MUC1基因疫苗诱导小鼠产生特异性体液及细胞免疫应答的作用,为壳聚糖联合MUC1基因疫苗用于胰腺癌的免疫治疗提供初步实验依据。方法 将30只小鼠平均分3组,分别接种壳聚糖-MUC1质粒、MUC1质粒及空质粒,通过ELISA法检测小鼠血清MUC1抗体生成情况,通过LDH释放法测定CTL对Capan-2细胞的杀伤活性。结果 接种壳聚糖-MUC1质粒后,小鼠血清抗MUC1抗体水平及CTL对Capan-2细胞的杀伤率均明显升高,与其它2组相比有显著性差异(P<0.05)。 结论 壳聚糖作为一种基因递送载体的MUC1基因疫苗能够有效诱导小鼠产生抗MUC1特异性抗体及产生杀伤MUC1+细胞的CTL,为壳聚糖联合MUC1基因疫苗用于胰腺癌的免疫治疗提供了初步的实验基础。     

胰腺癌生物治疗的新靶点:MUC1

胰腺癌预后很差，消耗着巨大的社会卫生资源。目前对胰腺癌的治疗仍缺乏有效的治疗手段。65％胰腺癌病人在诊断明确后半年内死亡，约90％的病人一年内死亡。探索新的有效的治疗方法，是目前急需解决的重要课题。生物治疗，尤其是肿瘤疫苗，作为具有临床应用潜力和价值的治疗手段，近年来开始得到了深入的研究。其关键是选择有效的治疗靶点，即寻找合适的肿瘤相关抗原（tumor associate antigen，TAA）。     

免疫增强型胰腺癌MUC1-VNTR-C144 DNA疫苗的构建

目的 构建免疫增强型胰腺癌MUC1-VNTR-C144 DNA疫苗.方法 在PeDNA 3.1-VNTR/Myc-his(+)A质粒靶基因之后插入乙型肝炎病毒核心抗原C144基因,构建MUC1-VNTR-C144 DNA疫苗.45只C57BL/6小鼠随机分3组,VC组接种PeDNA 3.1-VNTR-C144/Myc-his(+)A质粒,V组单纯接种PeDNA 3.1-VNTR/Myc-his(+)A质粒,C组单纯接种pcDNA3.1-C144/Myc-his(+)A质粒.4周后加强免疫1次.结果 VC组小鼠脾细胞毒性T淋巴细胞(CTL)的特异性杀伤率(46.7±8.2)%显著高于V组(34.8±3.1)%和C组(12.9±1.2)%(E/T=80/1,P<0.01).而CTL对未经VNTR合成肽孵育的靶细胞EIA-VNTR杀伤率较低(P<0.01),VU 3C6可抑制CTL对经VNTR合成肽孵育的靶细胞EIA-VNTR+的杀伤活性(P<0.01),表明具有VC组诱生的CTL应答依然保持VNTR特异性.VC组小鼠血清抗VNTR抗体等效浓度(2810.2±308.3)mg/L高于V组(2323.5±238.3)mg/L和C组(2130.9±138.5)mg/L,差异有统计学意义(P<0.01).结论 增强型胰腺癌MUC1-VNTR-C144 DNA疫苗所诱生的VNTR特异的CTL应答和抗体应答显著增强.     

新型胰腺癌MUC1 DNA疫苗的免疫原性研究

目的 研究所构建的新型胰腺癌MUC1 DNA疫苗的免疫原性.方法 采用三种优化策略共构建4个重组质粒,接种免疫C57BL/6J (H 2b)雌性小鼠.每2周加强免疫一次,共免疫3次.每次免疫前及处死小鼠前检测抗体和细胞因子.最后一次免疫结束后2周取脾,体外经VNTR合成肽刺激培养后进行杀伤性T淋巴细胞(cytotoxic T lymphocyte,CTL)杀伤功能分析.结果 pIRES2-EGFP3VNTR组、pIRES2-EGFP-3VNTR-CI-144组、pIRES2-EGFP-3VNTR-mIL-18组、pIRES2-EGFP-3VNTR-CI-144-mIL-18组的CTL特异杀伤率显著高于空载体对照组和生理盐水对照组,三重优化构建的质粒pIRES2-EGFP-3VNTR-mIL-18的CTL特异性杀伤率高于双重优化构建和单重优化构建的质粒.各优化构建质粒组的抗MUC1抗体OD值均显著高于对照组(P＜0.05),但组间差异无统计学意义.各优化构建质粒组的外周血IFN-γ均高于对照组(P＜0.05),且以含有IL-18优化策略的两组为高(P＜0.05).结论 所构建的优化重组质粒免疫小鼠后均可诱发抗原特异的CTL应答和抗体应答.C1-144和IL-18可增强疫苗的免疫原性.     

GM-CSF增强胰腺癌MUC1-VNTR核酸疫苗免疫效果的研究

目的研究 GM-CSF 作为免疫佐剂对胰腺癌 MUC1-VNTR 核酸疫苗免疫效果的增强作用。方法 45只 C57BL/6小鼠随机分3组,在胫前肌注射100μg/100μl质粒 DNA 生理盐水溶液(VG 组接种 pcDNA3.1-VNTR/Myc-his(+)A 质粒联用 rmGM-CSF,V 组单纯接种 pcDNA3.1-VNTR/Myc-his(+)A 质粒,G 组单纯接种 rmGM-CSF)进行免疫接种。4周后加强免疫1次,6周后取血清 ELISA 法测抗体,取脾细胞悬液体外以 VNTR 合成肽特异刺激,6d 后进行杀伤试验。结果VG 组小鼠脾细胞 CTL 的特异性杀伤率显著高于 V 组(P<0.01)和 G 组(P<0.01),表明 GM-CSF增强了疫苗所诱发的 CTL 应答。而 CTL 对未经 VNTR 合成肽孵育的靶细胞 EL4-VNTR~-杀伤较低(P<0.01),VU 3C6可抑制 CTL 对经 VNTR 合成肽孵育的靶细胞 EL4-VNTR~+的杀伤活性(P<0.01),表明具有 VG 组诱生的 CTL 应答依然保持 VNTR 特异性。VG 组小鼠血清抗 VNTR 抗体等效浓度(3119.0±110.6)μg/ml 高于Ⅴ组(2323.5±238.3)μg/ml 和 G 组(2023.9±169.0)μg/ml,差别极为显著(P<0.01)。结论 GM-CSF 可增强自行构建的胰腺癌 MUCl-VNTR 核酸疫苗所诱生的 VNTR 特异的 CTL 应答和抗体应答。     

免疫增强型胰腺癌MUC1-VNTR-C144 DNA疫苗的构建

Objective To construct new MUC1-VNTR-C144 DNA vaccine for pancreatic cancer and study both CTL responses and antibody responses specific to VNTR enhanced by the C144 gene. Methods DNA encoding the 144 amino acids of the N-terminus of HBV core gene was cloned and then inserted into the recombinant plasmid pcDNA3.1-VNTR/Myc-his( + ) A. C57BL/6 mice were randomly immunized intramusscularly into anterior tibialis muscle with 100 μg plasmid DNA encoding VNTR ( V group,n = 15) or VNTR/CI44 (VC group,n = 15). Mice injected with the plasmid pcDNA3.1-C144/ Myc-his( + ) A vector (C group, n = 15 ) were used as controls. Four weeks later, all mice were injected a-gain with the same solution as before. Two weeks after the last immunization, spleen cells were obtained and analyzed for cytotoxic activity 6 days after in vitro stimulation by the synthesized VNTR polypeptide (Calcein AM assay). Blood was collected for the ELISA assay of anti-VNTR antibodies. Results The specific CTL killing rate against VNTR polypeptide induced in the VC group (46.7±8.2)% was signifi-cantly higher than in the V group (34.8±3.1 ) % and the C group ( 12.9±1.2) % ( P < 0.01 ) ,indicating the C144 gene enhances the CTL response induced by the vaccine. However,CTL lysed much more VNTR positive EIA than VNTR negative EIA ( P < 0.01 ), while the anti-VNTR antibody VU 3C6 significantly restrained such activity ( P <0.01 ) ,which indicated the CTL response was specific to VNTR. The equiva-lent concentration of specific antibody against VNTR in the VC group (2810.2±308.3 ) mg/L was signif-icantly higher than the V group (2323.5±238.3) mg/L and the C group (2130.9±138.5) mg/L (P < 0.01 ),which implied that the C144 gene could enhance the antibody response induced by the vaccine. Conclusion The VNTR specific CTL response and antibody response induced in mice immunized with new MUCl-VNTR-C144 DNA vaccine for pancreatic cancer can be enhanced significantly by the C144 gene.     

免疫增强型胰腺癌MUC1-VNTR-C144 DNA疫苗的构建

Objective To construct new MUC1-VNTR-C144 DNA vaccine for pancreatic cancer and study both CTL responses and antibody responses specific to VNTR enhanced by the C144 gene. Methods DNA encoding the 144 amino acids of the N-terminus of HBV core gene was cloned and then inserted into the recombinant plasmid pcDNA3.1-VNTR/Myc-his( + ) A. C57BL/6 mice were randomly immunized intramusscularly into anterior tibialis muscle with 100 μg plasmid DNA encoding VNTR ( V group,n = 15) or VNTR/CI44 (VC group,n = 15). Mice injected with the plasmid pcDNA3.1-C144/ Myc-his( + ) A vector (C group, n = 15 ) were used as controls. Four weeks later, all mice were injected a-gain with the same solution as before. Two weeks after the last immunization, spleen cells were obtained and analyzed for cytotoxic activity 6 days after in vitro stimulation by the synthesized VNTR polypeptide (Calcein AM assay). Blood was collected for the ELISA assay of anti-VNTR antibodies. Results The specific CTL killing rate against VNTR polypeptide induced in the VC group (46.7±8.2)% was signifi-cantly higher than in the V group (34.8±3.1 ) % and the C group ( 12.9±1.2) % ( P < 0.01 ) ,indicating the C144 gene enhances the CTL response induced by the vaccine. However,CTL lysed much more VNTR positive EIA than VNTR negative EIA ( P < 0.01 ), while the anti-VNTR antibody VU 3C6 significantly restrained such activity ( P <0.01 ) ,which indicated the CTL response was specific to VNTR. The equiva-lent concentration of specific antibody against VNTR in the VC group (2810.2±308.3 ) mg/L was signif-icantly higher than the V group (2323.5±238.3) mg/L and the C group (2130.9±138.5) mg/L (P < 0.01 ),which implied that the C144 gene could enhance the antibody response induced by the vaccine. Conclusion The VNTR specific CTL response and antibody response induced in mice immunized with new MUCl-VNTR-C144 DNA vaccine for pancreatic cancer can be enhanced significantly by the C144 gene.     

免疫增强型胰腺癌MUC1-VNTR-C144 DNA疫苗的构建

Objective To construct new MUC1-VNTR-C144 DNA vaccine for pancreatic cancer and study both CTL responses and antibody responses specific to VNTR enhanced by the C144 gene. Methods DNA encoding the 144 amino acids of the N-terminus of HBV core gene was cloned and then inserted into the recombinant plasmid pcDNA3.1-VNTR/Myc-his( + ) A. C57BL/6 mice were randomly immunized intramusscularly into anterior tibialis muscle with 100 μg plasmid DNA encoding VNTR ( V group,n = 15) or VNTR/CI44 (VC group,n = 15). Mice injected with the plasmid pcDNA3.1-C144/ Myc-his( + ) A vector (C group, n = 15 ) were used as controls. Four weeks later, all mice were injected a-gain with the same solution as before. Two weeks after the last immunization, spleen cells were obtained and analyzed for cytotoxic activity 6 days after in vitro stimulation by the synthesized VNTR polypeptide (Calcein AM assay). Blood was collected for the ELISA assay of anti-VNTR antibodies. Results The specific CTL killing rate against VNTR polypeptide induced in the VC group (46.7±8.2)% was signifi-cantly higher than in the V group (34.8±3.1 ) % and the C group ( 12.9±1.2) % ( P < 0.01 ) ,indicating the C144 gene enhances the CTL response induced by the vaccine. However,CTL lysed much more VNTR positive EIA than VNTR negative EIA ( P < 0.01 ), while the anti-VNTR antibody VU 3C6 significantly restrained such activity ( P <0.01 ) ,which indicated the CTL response was specific to VNTR. The equiva-lent concentration of specific antibody against VNTR in the VC group (2810.2±308.3 ) mg/L was signif-icantly higher than the V group (2323.5±238.3) mg/L and the C group (2130.9±138.5) mg/L (P < 0.01 ),which implied that the C144 gene could enhance the antibody response induced by the vaccine. Conclusion The VNTR specific CTL response and antibody response induced in mice immunized with new MUCl-VNTR-C144 DNA vaccine for pancreatic cancer can be enhanced significantly by the C144 gene.     

免疫增强型胰腺癌MUC1-VNTR-C144 DNA疫苗的构建

Objective To construct new MUC1-VNTR-C144 DNA vaccine for pancreatic cancer and study both CTL responses and antibody responses specific to VNTR enhanced by the C144 gene. Methods DNA encoding the 144 amino acids of the N-terminus of HBV core gene was cloned and then inserted into the recombinant plasmid pcDNA3.1-VNTR/Myc-his( + ) A. C57BL/6 mice were randomly immunized intramusscularly into anterior tibialis muscle with 100 μg plasmid DNA encoding VNTR ( V group,n = 15) or VNTR/CI44 (VC group,n = 15). Mice injected with the plasmid pcDNA3.1-C144/ Myc-his( + ) A vector (C group, n = 15 ) were used as controls. Four weeks later, all mice were injected a-gain with the same solution as before. Two weeks after the last immunization, spleen cells were obtained and analyzed for cytotoxic activity 6 days after in vitro stimulation by the synthesized VNTR polypeptide (Calcein AM assay). Blood was collected for the ELISA assay of anti-VNTR antibodies. Results The specific CTL killing rate against VNTR polypeptide induced in the VC group (46.7±8.2)% was signifi-cantly higher than in the V group (34.8±3.1 ) % and the C group ( 12.9±1.2) % ( P < 0.01 ) ,indicating the C144 gene enhances the CTL response induced by the vaccine. However,CTL lysed much more VNTR positive EIA than VNTR negative EIA ( P < 0.01 ), while the anti-VNTR antibody VU 3C6 significantly restrained such activity ( P <0.01 ) ,which indicated the CTL response was specific to VNTR. The equiva-lent concentration of specific antibody against VNTR in the VC group (2810.2±308.3 ) mg/L was signif-icantly higher than the V group (2323.5±238.3) mg/L and the C group (2130.9±138.5) mg/L (P < 0.01 ),which implied that the C144 gene could enhance the antibody response induced by the vaccine. Conclusion The VNTR specific CTL response and antibody response induced in mice immunized with new MUCl-VNTR-C144 DNA vaccine for pancreatic cancer can be enhanced significantly by the C144 gene.     

免疫增强型胰腺癌MUC1-VNTR-C144 DNA疫苗的构建

Objective To construct new MUC1-VNTR-C144 DNA vaccine for pancreatic cancer and study both CTL responses and antibody responses specific to VNTR enhanced by the C144 gene. Methods DNA encoding the 144 amino acids of the N-terminus of HBV core gene was cloned and then inserted into the recombinant plasmid pcDNA3.1-VNTR/Myc-his( + ) A. C57BL/6 mice were randomly immunized intramusscularly into anterior tibialis muscle with 100 μg plasmid DNA encoding VNTR ( V group,n = 15) or VNTR/CI44 (VC group,n = 15). Mice injected with the plasmid pcDNA3.1-C144/ Myc-his( + ) A vector (C group, n = 15 ) were used as controls. Four weeks later, all mice were injected a-gain with the same solution as before. Two weeks after the last immunization, spleen cells were obtained and analyzed for cytotoxic activity 6 days after in vitro stimulation by the synthesized VNTR polypeptide (Calcein AM assay). Blood was collected for the ELISA assay of anti-VNTR antibodies. Results The specific CTL killing rate against VNTR polypeptide induced in the VC group (46.7±8.2)% was signifi-cantly higher than in the V group (34.8±3.1 ) % and the C group ( 12.9±1.2) % ( P < 0.01 ) ,indicating the C144 gene enhances the CTL response induced by the vaccine. However,CTL lysed much more VNTR positive EIA than VNTR negative EIA ( P < 0.01 ), while the anti-VNTR antibody VU 3C6 significantly restrained such activity ( P <0.01 ) ,which indicated the CTL response was specific to VNTR. The equiva-lent concentration of specific antibody against VNTR in the VC group (2810.2±308.3 ) mg/L was signif-icantly higher than the V group (2323.5±238.3) mg/L and the C group (2130.9±138.5) mg/L (P < 0.01 ),which implied that the C144 gene could enhance the antibody response induced by the vaccine. Conclusion The VNTR specific CTL response and antibody response induced in mice immunized with new MUCl-VNTR-C144 DNA vaccine for pancreatic cancer can be enhanced significantly by the C144 gene.     

免疫增强型胰腺癌MUC1-VNTR-C144 DNA疫苗的构建

Objective To construct new MUC1-VNTR-C144 DNA vaccine for pancreatic cancer and study both CTL responses and antibody responses specific to VNTR enhanced by the C144 gene. Methods DNA encoding the 144 amino acids of the N-terminus of HBV core gene was cloned and then inserted into the recombinant plasmid pcDNA3.1-VNTR/Myc-his( + ) A. C57BL/6 mice were randomly immunized intramusscularly into anterior tibialis muscle with 100 μg plasmid DNA encoding VNTR ( V group,n = 15) or VNTR/CI44 (VC group,n = 15). Mice injected with the plasmid pcDNA3.1-C144/ Myc-his( + ) A vector (C group, n = 15 ) were used as controls. Four weeks later, all mice were injected a-gain with the same solution as before. Two weeks after the last immunization, spleen cells were obtained and analyzed for cytotoxic activity 6 days after in vitro stimulation by the synthesized VNTR polypeptide (Calcein AM assay). Blood was collected for the ELISA assay of anti-VNTR antibodies. Results The specific CTL killing rate against VNTR polypeptide induced in the VC group (46.7±8.2)% was signifi-cantly higher than in the V group (34.8±3.1 ) % and the C group ( 12.9±1.2) % ( P < 0.01 ) ,indicating the C144 gene enhances the CTL response induced by the vaccine. However,CTL lysed much more VNTR positive EIA than VNTR negative EIA ( P < 0.01 ), while the anti-VNTR antibody VU 3C6 significantly restrained such activity ( P <0.01 ) ,which indicated the CTL response was specific to VNTR. The equiva-lent concentration of specific antibody against VNTR in the VC group (2810.2±308.3 ) mg/L was signif-icantly higher than the V group (2323.5±238.3) mg/L and the C group (2130.9±138.5) mg/L (P < 0.01 ),which implied that the C144 gene could enhance the antibody response induced by the vaccine. Conclusion The VNTR specific CTL response and antibody response induced in mice immunized with new MUCl-VNTR-C144 DNA vaccine for pancreatic cancer can be enhanced significantly by the C144 gene.     

免疫增强型胰腺癌MUC1-VNTR-C144 DNA疫苗的构建

Objective To construct new MUC1-VNTR-C144 DNA vaccine for pancreatic cancer and study both CTL responses and antibody responses specific to VNTR enhanced by the C144 gene. Methods DNA encoding the 144 amino acids of the N-terminus of HBV core gene was cloned and then inserted into the recombinant plasmid pcDNA3.1-VNTR/Myc-his( + ) A. C57BL/6 mice were randomly immunized intramusscularly into anterior tibialis muscle with 100 μg plasmid DNA encoding VNTR ( V group,n = 15) or VNTR/CI44 (VC group,n = 15). Mice injected with the plasmid pcDNA3.1-C144/ Myc-his( + ) A vector (C group, n = 15 ) were used as controls. Four weeks later, all mice were injected a-gain with the same solution as before. Two weeks after the last immunization, spleen cells were obtained and analyzed for cytotoxic activity 6 days after in vitro stimulation by the synthesized VNTR polypeptide (Calcein AM assay). Blood was collected for the ELISA assay of anti-VNTR antibodies. Results The specific CTL killing rate against VNTR polypeptide induced in the VC group (46.7±8.2)% was signifi-cantly higher than in the V group (34.8±3.1 ) % and the C group ( 12.9±1.2) % ( P < 0.01 ) ,indicating the C144 gene enhances the CTL response induced by the vaccine. However,CTL lysed much more VNTR positive EIA than VNTR negative EIA ( P < 0.01 ), while the anti-VNTR antibody VU 3C6 significantly restrained such activity ( P <0.01 ) ,which indicated the CTL response was specific to VNTR. The equiva-lent concentration of specific antibody against VNTR in the VC group (2810.2±308.3 ) mg/L was signif-icantly higher than the V group (2323.5±238.3) mg/L and the C group (2130.9±138.5) mg/L (P < 0.01 ),which implied that the C144 gene could enhance the antibody response induced by the vaccine. Conclusion The VNTR specific CTL response and antibody response induced in mice immunized with new MUCl-VNTR-C144 DNA vaccine for pancreatic cancer can be enhanced significantly by the C144 gene.     

免疫增强型胰腺癌MUC1-VNTR-C144 DNA疫苗的构建

Objective To construct new MUC1-VNTR-C144 DNA vaccine for pancreatic cancer and study both CTL responses and antibody responses specific to VNTR enhanced by the C144 gene. Methods DNA encoding the 144 amino acids of the N-terminus of HBV core gene was cloned and then inserted into the recombinant plasmid pcDNA3.1-VNTR/Myc-his( + ) A. C57BL/6 mice were randomly immunized intramusscularly into anterior tibialis muscle with 100 μg plasmid DNA encoding VNTR ( V group,n = 15) or VNTR/CI44 (VC group,n = 15). Mice injected with the plasmid pcDNA3.1-C144/ Myc-his( + ) A vector (C group, n = 15 ) were used as controls. Four weeks later, all mice were injected a-gain with the same solution as before. Two weeks after the last immunization, spleen cells were obtained and analyzed for cytotoxic activity 6 days after in vitro stimulation by the synthesized VNTR polypeptide (Calcein AM assay). Blood was collected for the ELISA assay of anti-VNTR antibodies. Results The specific CTL killing rate against VNTR polypeptide induced in the VC group (46.7±8.2)% was signifi-cantly higher than in the V group (34.8±3.1 ) % and the C group ( 12.9±1.2) % ( P < 0.01 ) ,indicating the C144 gene enhances the CTL response induced by the vaccine. However,CTL lysed much more VNTR positive EIA than VNTR negative EIA ( P < 0.01 ), while the anti-VNTR antibody VU 3C6 significantly restrained such activity ( P <0.01 ) ,which indicated the CTL response was specific to VNTR. The equiva-lent concentration of specific antibody against VNTR in the VC group (2810.2±308.3 ) mg/L was signif-icantly higher than the V group (2323.5±238.3) mg/L and the C group (2130.9±138.5) mg/L (P < 0.01 ),which implied that the C144 gene could enhance the antibody response induced by the vaccine. Conclusion The VNTR specific CTL response and antibody response induced in mice immunized with new MUCl-VNTR-C144 DNA vaccine for pancreatic cancer can be enhanced significantly by the C144 gene.     

免疫增强型胰腺癌MUC1-VNTR-C144 DNA疫苗的构建

Objective To construct new MUC1-VNTR-C144 DNA vaccine for pancreatic cancer and study both CTL responses and antibody responses specific to VNTR enhanced by the C144 gene. Methods DNA encoding the 144 amino acids of the N-terminus of HBV core gene was cloned and then inserted into the recombinant plasmid pcDNA3.1-VNTR/Myc-his( + ) A. C57BL/6 mice were randomly immunized intramusscularly into anterior tibialis muscle with 100 μg plasmid DNA encoding VNTR ( V group,n = 15) or VNTR/CI44 (VC group,n = 15). Mice injected with the plasmid pcDNA3.1-C144/ Myc-his( + ) A vector (C group, n = 15 ) were used as controls. Four weeks later, all mice were injected a-gain with the same solution as before. Two weeks after the last immunization, spleen cells were obtained and analyzed for cytotoxic activity 6 days after in vitro stimulation by the synthesized VNTR polypeptide (Calcein AM assay). Blood was collected for the ELISA assay of anti-VNTR antibodies. Results The specific CTL killing rate against VNTR polypeptide induced in the VC group (46.7±8.2)% was signifi-cantly higher than in the V group (34.8±3.1 ) % and the C group ( 12.9±1.2) % ( P < 0.01 ) ,indicating the C144 gene enhances the CTL response induced by the vaccine. However,CTL lysed much more VNTR positive EIA than VNTR negative EIA ( P < 0.01 ), while the anti-VNTR antibody VU 3C6 significantly restrained such activity ( P <0.01 ) ,which indicated the CTL response was specific to VNTR. The equiva-lent concentration of specific antibody against VNTR in the VC group (2810.2±308.3 ) mg/L was signif-icantly higher than the V group (2323.5±238.3) mg/L and the C group (2130.9±138.5) mg/L (P < 0.01 ),which implied that the C144 gene could enhance the antibody response induced by the vaccine. Conclusion The VNTR specific CTL response and antibody response induced in mice immunized with new MUCl-VNTR-C144 DNA vaccine for pancreatic cancer can be enhanced significantly by the C144 gene.     

胰腺癌MUC1-VNTR核酸疫苗的构建和体外转染

目的 探索胰腺癌MUC1-VNTR核酸疫苗的构建，以进一步研究对胰腺癌的治疗作用。方法 首先对VNTR编码基因进行了重组设计，氨基端插入人单核细胞趋化蛋白Ⅰ信号肽基因序列，起始码前插入KOZAK促真核翻译序列。将人工合成的重组人VNTR基因定向克隆入真核表达载体pcDNA3，1／Myc-his（＋）A质粒的MCS中。将通过测序鉴定的含有准确插入序列的pcDNA3．1-VNTR／Myc-his（十）A重组质粒转染C087细胞，进行体外转染实验和Westernblot检测VNTR在细胞内外的表达。结果自转化平板筛选的阳性克隆通过测序鉴定，pcDNA3．1-VNTR／Myc-his（＋）A重组质粒含有完整的阅读框架和目的基因。重组质粒可在C087细胞内外表达VNTR，以胞内为主。所表达的VNTR分子量比人工合成的VNTR多肽大。结论 自行构建的胰腺癌MUC1-VNTR核酸疫苗序列准确，可在哺乳动物细胞中表达VNTR。     

重视胰腺癌新辅助治疗的临床研究

数十年来针对胰腺癌的临床研究有诸多热点课题。如淋巴清扫范围、辅助治疗、联合血管切除、切缘的判断标准等,随着对胰腺癌临床及生物学行为的认识不断深入,手术切除率及手术安全性有了较大提高,但治疗效果并无显著改善,胰腺癌患者总体中位生存期为5～8个月,5年生存率仅为5％左右。近年来,人们对辅助治疗改善患者预后的意义已有基本共识,临床应用日益规范,针对新辅助治疗的临床研究渐为热点。     

新辅助化疗后门静脉系统全置换治疗局部进展期胰腺癌一例报告

患者女性,65岁,局部进展期胰腺癌伴门静脉、肠系膜上静脉和肠系膜上动脉侵犯,经6个周期奥沙利铂＋氟尿嘧啶＋伊立替康＋亚叶酸钙(FOLFIRINOX)新辅助化疗后,肿瘤评估为部分缓解,但仍属于局部进展期胰腺癌。患者行动脉优先入路、全门静脉系统切除、异体血管置换重建的扩大胰十二指肠切除术,同时运用异体血管体外成形技术。患者...     

针对MUC1及其启动子的恶性肿瘤生物靶向治疗(文献综述)

MUC1是属于mucins家族的一种糖蛋白 ,主要存在于某些上皮细胞 ,癌变后 ,其表达可达正常时的 1 0倍以上 ,且糖链变短 ,使核心肽暴露 ,显示出较高的癌特异性和免疫原性。另外 ,MUC1启动子所调控的基因表达具有较高的组织和细胞特异性。目前 ,针对它们已开展了一些恶性肿瘤的生物靶向治疗 ,有的已进入临床试验阶段     

胰腺癌的早期诊断和外科治疗

目的：总结胰腺癌的诊治经验。方法：对250例胰腺癌的诊治进行回顾性分析。结果：（1）胰腺癌首发症状并非无痛性黄疸，而主要为上腹部胀痛不适，纳差，明显乏力和消瘦等。B超、CT和ERCP等影像学检查可发现胰腺癌的间接征象（胆道扩张、胰管狭窄、移位和增粗等）。（2）胰腺残端闭锁式Child吻合加空肠输入袢减压，术后并发症和手术死亡率明显降低。结论：（1）对高危病人提高警惕。首选B超，联合应用CT和ERCP可提高胰腺癌早期诊断。（2）PD改良式Child消化道重建术可降低并发症发生率和手术死亡率。     

固定剂量率输注吉西他滨治疗晚期胰腺癌

1.背景目前胰腺癌发病率呈逐年升高的趋势,早期难以诊断,确诊时约80%以上的患者属于晚期,已无手术切除的机会,化学治疗便成为晚期胰腺癌的主要治疗手段之一[1-2]。1997年发表的一个126例III期胰腺癌患者的随机临床研究[3],首次确定吉西他滨单药化疗优于5-FU单药,从此吉西他滨30