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Ets1 as a marker of malignant potential in gastric carcinoma 被引量:8
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作者 YongYu Yi-ChuZhang +4 位作者 Wen-ZhuZhang Li-SongShen PaulHertzog TrevorJWilson Da-KangXu 《World Journal of Gastroenterology》 SCIE CAS CSCD 2003年第10期2154-2159,共6页
AIM: Ets1 proto-oncogene is a transcription factor involved in the activation of several genes of tumor invasion and metastasis. We aimed to determine the relationship between the extent and intensity of Ets1 expressi... AIM: Ets1 proto-oncogene is a transcription factor involved in the activation of several genes of tumor invasion and metastasis. We aimed to determine the relationship between the extent and intensity of Ets1 expression and patients'' clinicopathological factors in gastric carcinoma. METHODS: Immunohistochemical analysis was performed for gastric tumor paraffin-embedded sections, followed by image analysis. RESULTS: Ets1 was not expressed in the normal gastric epithelium and its surrounding cells. The percentage of Ets1 expressing cells detected increased significantly in both epithelial tumor and stromal cells from high T classification, lymph node metastasis positive, clinical advanced-stage groups (P<0.05). The level of Etsl staining in epithelial tumor cells also reflected the degree of cell differentiation. The percentage of epithelial and stromal cells expressing Ets1 was significantly correlated with the presence of lymph node metastasis (P=0.014 and P=0.O01 respectively). Ets1 expression was not observed in tissue samples from patients with benign gastric ulcers.CONCLUSION: Ets1 protein expression in epithelial tumor cells reflects the degree of differentiation, and the percentage of Etsl positive tumor and stromal cells correlates with lymph node metastasis. Thus Etsl is a valuable marker of malignant potential in terms of invasiveness and metastasis of gastric carcinoma. It is also possible that inhibition of Etsl is a potential avenue for therapy in gastric cancer. 展开更多
关键词 胃癌 致癌基因 肿瘤侵袭 肿瘤转移 免疫组织化学 ETS1 基质细胞
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大鼠胎脑神经干细胞HPRT基因的敲除 被引量:1
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作者 李雪玲 扈廷茂 John R Morrison 《Acta Genetica Sinica》 SCIE CAS CSCD 北大核心 2004年第1期51-56,共6页
根据已知大鼠次黄嘌呤鸟嘌呤磷酸核糖转移酶 (HypoxanthineGuaninePhosphoribosylTransferase ,HPRT)基因的外显子序列 ,从大鼠HPRT基因组DNA序列的细菌人工染色体 (BacterialArtificialChromosome ,BAC)中用酶切和PCR方法分别分离得到... 根据已知大鼠次黄嘌呤鸟嘌呤磷酸核糖转移酶 (HypoxanthineGuaninePhosphoribosylTransferase ,HPRT)基因的外显子序列 ,从大鼠HPRT基因组DNA序列的细菌人工染色体 (BacterialArtificialChromosome ,BAC)中用酶切和PCR方法分别分离得到用于构建基因敲除载体的 3 0kb的 5′长臂 (LongArm ,LA)和 1 7kb的 3′短臂 (ShortArm ,SA) ,并分别克隆到pSL1180和pCR2 1中。进一步构建大鼠HPRT基因打靶载体———pKO HPRT ,经酶切鉴定后的大鼠HPRT基因敲除载体用NotⅠ酶切使其线性化 ,经溴乙锭、正丁醇、酚、酚 /氯仿提纯后 ,将终浓度调至 1μg μl。在FuGene 6转染试剂的作用下转染培养 2 4h的第二代大鼠胎脑神经干细胞 (RatFetalNeuralStemCells,rFNSCs)。转染后的细胞用 80 μg mlG4 18和 0 2 μmol L的Ganc全培养液筛选 ,2w后将存活细胞进行悬浮培养 ,使细胞形成球形物 ,挑选单个的球形物进行单克隆增殖 ,其中一部分细胞 (约 2~ 3× 10 3)用裂解液处理 ,取上清用于PCR检测 ,大部分细胞 (5× 10 7)用于DNA和RNA的提取 ,进行Southernbolt和RT PCR检测 ,剩余细胞冷冻保存。最后 1次实验共分离培养了 32个rFNSCs单克隆 ,其中 3个单克隆 (9 3% )经PCR、Southernbolt和RT 展开更多
关键词 大鼠胎脑神经干细胞 基因敲除 次黄嘌呤鸟嘌呤磷酸核糖转移酶基因
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