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Objective To obtain the dendritic cells ( DC)-based vaccine modified by adenovirus containing MUC4 gene , and evaluate the anti-tumor efficacy of DC vaccine to pancreatic tumor cells. Meth-ods The mRNA sequence of tumor associated antigen, MUC4, was obtained from NCBI, and MUC4 se-quence was acquired through the restriction enzyme sites and over lap PCR, then subcloned into adenovirus plasmid to create recombinant adenovirus ( rAd-MUC4) . The DCs were infected by rAd-MUC4 virus and then stimulated the lymph cells from the same donor to induce MUC4 specific cytotoxicity T lympbocytes ( CTL) . The efficacy of CTL was analyzed by LDH releasing assay. Elispot was used to detect the IFN-γ release. Results The recombinant adenovirus containing MUC4 ( sv12) gene was obtained. The MUC4-induced CTL could specifically kill the Capan-1 pancreatic tumor cells [ ( 13. 7±6.0)% , ( 21.4± 4. 7)% , (36.1±9. 5)% at ratios of 10: ,20: ,40: ] , higher than MCF-7 and Bxpc-3 cells respectively, P < 0. 05. The spots number of CTL induced by rAd-MUC4 was ( 139.1±23.3) , more than GFP and PBS control group,P<0.05. Conclusion The Muc4 gene modified DC vaccine could induce the proliferation of CTL, which provided a significant cytotoxicity to HLA-matched MUC4 positive tumor cell lines in vitro. 相似文献
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目的:根据肿瘤干细胞学说理论,探讨胰腺癌耐药的新机制。方法:通过胰腺癌肿瘤干细胞表面标记途径和侧群(side population,SP)细胞途径,分离出人胰腺癌细胞株PANC-1中的SP/NSP(非SP)细胞及CD44+CD24+/CD44-CD24-细胞亚群,用MTT检测上述各亚群细胞在体外对化疗药物耐受的差异,用AnnexinV-PI双染法检测2种肿瘤细胞的抗凋亡能力,并采用实时荧光定量PCR检测两者耐药基因ABCG2、ABCB1和PLK-1表达的差异。结果:人胰腺癌细胞株PANC-1中的SP细胞比例为(7.64±0.96)%,CD44+CD24+细胞比例为(2.60±0.96)%。相对于NSP和CD44-CD24-细胞而言,SP和CD44+CD24+细胞具有更强的化疗耐受能力(P0.01)和抗凋亡能力(P0.01);荧光定量RT-PCR结果均提示,SP和CD44+CD24+细胞高表达耐药基因ABCG2、ABCB1和PLK1。结论:人胰腺癌细胞株PANC-1中SP及CD44+CD24+细胞具有更强的化疗耐受能力,研究胰腺癌肿瘤干细胞可为克服胰腺癌化疗敏感性差的现状提供新的实验基础和理论依据。 相似文献
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目的:探讨Hedgehog信号通路蛋白在人胰腺癌吉西他滨耐药细胞株SW1990中的表达,为克服胰腺癌获得性耐药提供实验基础。方法:采用浓度梯度递增法建立人胰腺癌SW1990耐药株,采用噻唑蓝法测定SW1990亲代与耐药细胞IC50。实时荧光定量PCR检测亲代与耐药细胞mRNA中hedgehog信号通路成员Shh、SMO、Gli-1的表达差异。Western印迹法检测亲代与耐药细胞中上述蛋白质的表达。结果:人胰腺癌耐药株SW1990的IC50从亲代的(3.1±0.2)μmol/L提高到(232.2±12.3)μmol/L。荧光定量PCR结果显示耐药株中Shh、Gli-1的表达提高了(12.07±1.71)倍和(4.15±0.42)倍。亲代SW1990中未检测到SMO表达,而耐药细胞中却可以检测到SMO的表达。Western印迹结果同样显示,人胰腺癌SW1990耐药细胞株中高表达上述蛋白质。结论:人胰腺癌耐药株中高表达部分hedgehog信号通路蛋白。针对hedgehog信号通路的靶向治疗可能为克服胰腺癌耐药提供新的理论基础。 相似文献
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我们通过构建小鼠半乳糖凝集素1( mGalectin-1)基因慢病毒过表达载体并感染原代培养的小鼠胰腺星状细胞( mPSCs),探讨mGalectin-1与mPSCs增殖之间的关系.
一、材料和方法
1.材料:293T细胞株由本实验室保存,采用腹主动脉灌注法分离原代mPSCs并培养鉴定[1],慢病毒表达载体pHAGE及辅助包装元件载体( psPAX2,pMD2.G)由南京医科大学现代病原生物学重点实验室卢春教授提供. 相似文献
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目的 探讨老年急性胰腺炎(AP)患者的临床特征和诊疗效果。方法 回顾性地分析南京医科大学第一附属医院胰腺中心2012年1月至2014年12月期间收治的164例老年AP患者(老年组,年龄≥60岁)临床特征和疗效,并与同期收治的309例非老年AP患者(对照组,年龄<60岁)进行对比分析。结果 老年组AP的主要病因为胆道疾病,其次为高脂血症,老年组胆源性AP发生率明显高于对照组(84.15% vs 59.55%,P<0.001),高脂血症性AP发生率明显低于对照组(9.14% vs 31.07%,P<0.001)。老年组和对照组主要全身并发症均为脏器功能衰竭(20.12% vs 18.77%,P>0.05),但老年组全身感染和持续性全身炎症反应综合征发生率明显高于对照组,两组间差异具有统计学意义(P<0.05),两组间局部并发症发生率无统计学差异(P>0.05)。老年组重症急性胰腺炎(SAP)发生率与对照组相当(18.90% vs 18.77%),但病死率明显高于对照组(7.93% vs 3.56%,P<0.05)。结论 老年AP患者合并基础疾病多,易发生全身并发症,发展为SAP后病死率高,临床应予以早期诊断和有效治疗,可改善老年AP患者的预后。 相似文献
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目的:以围手术期参数为指标,对单中心腹腔镜下胰十二指肠切除术(laparoscopic pancreaticoduodenectomy,LPD)的学习曲线进行回顾性分析。方法:回顾性分析了2015年1月—2019年3月,南京医科大学第一附属医院行LPD的92例患者,对其围手术期参数进行统计分析。并以31、30、31例为界,区分3个学习曲线完成度,并进行分阶段参数的比较。结果:按照学习曲线划分,其中第1阶段患者中位手术时间为8.7(5.0,7.0)h,平均住院时间15.0(1.02,28.0)d,胰瘘发生率51.6%。第2阶段患者中位手术时间为7.5(7.0,10.0)h,平均住院时间12.5(9.0,16.0)d,胰瘘发生率23.3%。第3阶段患者中位手术时间为8.0(8.0,9.0)h,中位住院时间16.0(12.0,23.0)d,胰瘘发生率12.9%。手术时间和住院时间(length of stay,LOS)3个阶段之间差异无统计学意义(P > 0.05);3个阶段的胰瘘率差异有统计学意义(P < 0.05)。结论:LPD技术挑战高,具有很长的学习曲线,在累积92例后仍未达到平台期,但围手术期参数已随病例积累而改善。 相似文献
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Objective To obtain the dendritic cells ( DC)-based vaccine modified by adenovirus containing MUC4 gene , and evaluate the anti-tumor efficacy of DC vaccine to pancreatic tumor cells. Meth-ods The mRNA sequence of tumor associated antigen, MUC4, was obtained from NCBI, and MUC4 se-quence was acquired through the restriction enzyme sites and over lap PCR, then subcloned into adenovirus plasmid to create recombinant adenovirus ( rAd-MUC4) . The DCs were infected by rAd-MUC4 virus and then stimulated the lymph cells from the same donor to induce MUC4 specific cytotoxicity T lympbocytes ( CTL) . The efficacy of CTL was analyzed by LDH releasing assay. Elispot was used to detect the IFN-γ release. Results The recombinant adenovirus containing MUC4 ( sv12) gene was obtained. The MUC4-induced CTL could specifically kill the Capan-1 pancreatic tumor cells [ ( 13. 7±6.0)% , ( 21.4± 4. 7)% , (36.1±9. 5)% at ratios of 10: ,20: ,40: ] , higher than MCF-7 and Bxpc-3 cells respectively, P < 0. 05. The spots number of CTL induced by rAd-MUC4 was ( 139.1±23.3) , more than GFP and PBS control group,P<0.05. Conclusion The Muc4 gene modified DC vaccine could induce the proliferation of CTL, which provided a significant cytotoxicity to HLA-matched MUC4 positive tumor cell lines in vitro. 相似文献
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胰腺癌导致的神经改变包括胰腺支配神经的侵袭性改变和非侵袭性改变,即嗜神经侵袭与神经重塑.嗜神经侵袭是指神经三层包膜(神经外膜、神经束膜、神经内膜)中任意一层被胰腺癌细胞侵犯或癌细胞在神经外周包绕超过1/3周长,是癌细胞与神经纤维的直接接触[1-2].神经重塑是指胰腺组织癌变时其支配神经出现神经肥大、密度增加以及神经炎症... 相似文献