热休克转录因子1在人肝细胞癌中的表达及转录活性 滕洪飞.docxVIP

热休克转录因子1在人肝细胞癌中的表达及转录活性 滕洪飞 热休克转录因子1在人肝细胞癌中的表达及转录活性lowbar;滕洪飞 热休克转录因子1在人肝细胞癌中的表达及转录活性 宋振顺李煜环范子扬 HSF1）表达量及观察人肝细胞癌组织及对应癌旁组织中热休克转录因子1（heat shock transcription factor 1， 免疫组织化学法及Western blot 转录活性。方法用免疫组织化学方法及Western blot 方法测定30例人肝细胞癌组织及对应癌旁组织中HSF1的表达量及磷酸 化水平，并比较HSF1的表达量及磷酸化水平在肝细胞癌组织及对应癌旁组织中的差异。结果癌旁组织。结论 HSF1在人肝细胞癌中的表达量及转录活性明显增高。 结果显示：①人肝细胞癌组织中HSF1的表达量明显高于对应癌旁组织；②人肝细胞癌组织中HSF1的磷酸化水平明显高于对应 热休克转录因子1 Expression and Transcriptional Activity of HSF1in Human Hepatocellular Carcinoma． Teng Hongfei ，Song Zhenshun ，Li Yuhuan ，Fan Ziyang．Department of Hepatobiliary and Pancreato －Splenic Surgery ，Xijing Hospital ，The Fourth Military Medical University ，Shaanxi 710032，China Abstract Objective To observe the expression and transcriptional activity of HSF1in human hepatocellular carcinoma．Methods The results of immunohisto-The expression and phosphorylation of HSF1in several cases of human hepatocelluar carcinoma tissues and adjacent tissues was detected by immunohistochemical and Western Blot methods．The differences was analyzed between them．Ｒesults chemical and Western blot suggest．expression of HSF1in human hepatocelluar carcinoma tissues was higher than that in adjacent tissues ，the level of phosphorylation of HSF1in human hepatocelluar carcinoma tissues was higher than that in adjacent tissues too．Conclusion Expression and transcriptional activity of HSF1in human hepatocellular carcinoma tissues were higher than those in adjacent tissues． Key words HSF1；HCC ；Expression ；Phosphorylation 10%甲醛稀释液固定做冷冻切片，组织切片厚度为4 8mm ，5min ，室温下自然晾干。检测HSF1表达量及磷酸化时使用SP 法。在癌及癌旁组织切片上加入1? 100稀释的兔抗人HSF1单克隆抗体于4? 孵育过夜。PBS 洗涤5min ? 3次，加37? 孵育30min 后再加入辣入生物素标记的山羊抗兔二抗， 37? 再次孵育30min ，PBS 洗涤5min ? 3根酶标记链霉卵白素， 次，加入显色剂DAB 显色，自来水冲洗后苏木静衬染。（2）HSF1磷酸化水平：癌及癌旁组织切片上加入1? 100稀释的兔抗人p －HSF1单克隆抗体，其余步骤同前。 3．测定HSF1的表达量：将新鲜的人肝细胞癌组织及对加入ＲIPA裂解液1ml 应癌旁组织各100mg 冰浴中充分匀浆， 4? 条件下12000r /min离心30min ，充分裂解10min ，收集上清液并在相同条件下再次离心后取上清液；BCA 法