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1 材料和方法1.1 材料手术切除 PHC 38例,男26例,女12例,年龄32岁～71岁,平均47.4岁,术前均未放疗,标本均于离体后数小时内采集,液氮速冻后,转至-80℃冰箱贮存备用,所有病例均经病理证实.1.2 方法端粒酶提取:每例标本取原发性肝癌及癌旁组织各100 mg,置组织研蘑器内研成粉沫,再转至1.5 mL 小管中,立即加入预冷的裂解液200 μL(10mmol/L Tris-Hcl PH7.5);1 mmol/L MgCl_2;1mmol/L UTT,1 mmol/L EGTA,5 mmol/L二硫基乙醇,5 g/L CHAPS,100mL/L 甘油混均后30min 0℃裂解,4℃离心30min,吸取上清进行蛋白质含量测定,置-80℃
1 Materials and methods 1.1 Materials 38 cases of PHC were surgically removed, including 26 males and 12 females, aged 32 to 71 years, with an average of 47.4 years. No radiotherapy was performed before surgery. Specimens were collected within hours of being isolated. After quick-freezing, transfer to -80°C refrigerator for storage. All cases were confirmed by pathology. 1.2 Methods Telomerase extraction: 100 mg of primary hepatocellular carcinoma and paracancerous tissue were collected from each sample. The powder was then transferred to a 1.5 mL vial and 200 μL (10 mmol/L Tris-Hcl pH 7.5) of pre-chilled lysate was immediately added; 1 mmol/L MgCl 2 ; 1 mmol/L UTT, 1 mmol/L EGTA, 5 Dimethyl Ethanol, 5 g/L CHAPS, 100 mL/L glycerol mixed at 30°C 0°C, centrifuged at 4°C for 30min, aspirate supernatant to determine protein content, set -80°C