AIM To investigate the expression of perforin and fas ligand (fas L) of tumor infiltrating lymphocytes (TILs) in human hepatocellular carcinoma (HCC). METHODS By in situ hybridization and immunohistochemistry...AIM To investigate the expression of perforin and fas ligand (fas L) of tumor infiltrating lymphocytes (TILs) in human hepatocellular carcinoma (HCC). METHODS By in situ hybridization and immunohistochemistry, the perforin and fas L gene expression of TILs was studied in 20 HCC cases. RESULTS Positive expression of perforin and fas L genes was detected in 16 HCC cases. One patient had expression of perforin and fas L genes in the majority of TILs and survived 1 5 years after tumor resection without HCC relapse. This seems that the presence of a large number of activated T cells might be beneficial for the antitumor immunity. In other cases, less than 10% of TILs were able to express perforin and fas L genes. CONCLUSION Although there were a number of T cells in HCC, only few of them were immunoactive and able to kill tumor cells. It seems important to promote further proliferation of these activated T cells in vitro or in vivo .展开更多
用植物凝集素激活正常人单核细胞 ,诱导 Fas L m RNA表达 ,提取总 RNA,经逆转录多聚酶链反应( RT- PCR)筛选 Fas L基因片段。定向克隆入质粒 p SPT1 9,多克隆位点获取重组质粒 ,经 DNA测序后体外转录制备 Dig- Fas L c RNA探针。收集经...用植物凝集素激活正常人单核细胞 ,诱导 Fas L m RNA表达 ,提取总 RNA,经逆转录多聚酶链反应( RT- PCR)筛选 Fas L基因片段。定向克隆入质粒 p SPT1 9,多克隆位点获取重组质粒 ,经 DNA测序后体外转录制备 Dig- Fas L c RNA探针。收集经病理证实的肺癌及癌周正常肺组织新鲜标本 ,用液氮速冻 ,4%多聚甲醛固定。在冰冻切片上以 c RNA探针原位杂交进行 Fas L m RNA表达分布的观察。结果 :在鳞癌、腺癌和小细胞肺癌的癌组织及癌旁肺组织中均可检出 Fas L的杂交信号。镜下见鳞癌组织中 Fas展开更多
目的:探索应用细菌双杂交系统在细胞外蛋白质之间相互作用的可行性。方法:应用Stratagene BacterioMatch two hy-brid system,以pTRG构建编码TRAIL细胞外可溶性区域序列融合表达质粒;以pBT构建编码DR4、DR5、OPG等的细胞外区域序列融合...目的:探索应用细菌双杂交系统在细胞外蛋白质之间相互作用的可行性。方法:应用Stratagene BacterioMatch two hy-brid system,以pTRG构建编码TRAIL细胞外可溶性区域序列融合表达质粒;以pBT构建编码DR4、DR5、OPG等的细胞外区域序列融合表达质粒。重组质粒分别相应共转化报告菌株XL1-Blue MRF,报道基因产物以抗羧苄青霉素和β-半乳糖苷酶活性作为转录激活的指标。结果:pTRG-TRAIL与pBT-DR4,pTRG-TRAIL与pBT-DR5及对照组分别共转化后在高浓度羧苄青霉素(500μg/ml以上)筛选下宿主菌呈转录激活状态,pTRG-TRAIL与pBT-OPG组共转化后在低浓度羧苄青霉素(250μg/ml以下)筛选下宿主菌呈转录激活状态,用X-Gal-PETG平皿可以进一步验证阳性克隆。对照组pTRG-TRAIL与pBT-LGF2,pBT-DR4、pBT-DR5或pBT-OPG与pTRG-Gal114组共转化后呈阴性。结论:所用的双杂交序列皆为编码这些蛋白质的细胞外区域,而这些蛋白质本身的相互作用已有其他报道验证,所以细菌双杂交系统可以用于细胞外蛋白质的相互作用研究。展开更多
文摘AIM To investigate the expression of perforin and fas ligand (fas L) of tumor infiltrating lymphocytes (TILs) in human hepatocellular carcinoma (HCC). METHODS By in situ hybridization and immunohistochemistry, the perforin and fas L gene expression of TILs was studied in 20 HCC cases. RESULTS Positive expression of perforin and fas L genes was detected in 16 HCC cases. One patient had expression of perforin and fas L genes in the majority of TILs and survived 1 5 years after tumor resection without HCC relapse. This seems that the presence of a large number of activated T cells might be beneficial for the antitumor immunity. In other cases, less than 10% of TILs were able to express perforin and fas L genes. CONCLUSION Although there were a number of T cells in HCC, only few of them were immunoactive and able to kill tumor cells. It seems important to promote further proliferation of these activated T cells in vitro or in vivo .
文摘用植物凝集素激活正常人单核细胞 ,诱导 Fas L m RNA表达 ,提取总 RNA,经逆转录多聚酶链反应( RT- PCR)筛选 Fas L基因片段。定向克隆入质粒 p SPT1 9,多克隆位点获取重组质粒 ,经 DNA测序后体外转录制备 Dig- Fas L c RNA探针。收集经病理证实的肺癌及癌周正常肺组织新鲜标本 ,用液氮速冻 ,4%多聚甲醛固定。在冰冻切片上以 c RNA探针原位杂交进行 Fas L m RNA表达分布的观察。结果 :在鳞癌、腺癌和小细胞肺癌的癌组织及癌旁肺组织中均可检出 Fas L的杂交信号。镜下见鳞癌组织中 Fas
