肝癌细胞中端粒酶活性的检测

THE DETECTION OF TELOMERASE ACTIVITY IN HEPATOCARCINOMA

目的 本实验为检测肝癌组织中是否有端粒酶活性，端粒酶活性与肝癌临床检测指标ＡＦＰ及高低转移潜能是否相关而设计。 方法 本实验用ＴＲＡＰ法从组织中抽提端粒酶，经反转录并ＰＣＲ 扩增后用地高辛标记的探针作斑点印迹杂交（ 肝癌标本部分） 和ＰＡＧ电泳加银染法显色（ 人肝癌高低转移裸鼠模型） 。 结果 本实验检测了１１ 例肝癌患者有肝癌组织和癌周组织的端粒酶活性，结果肝癌组织中端粒酶活性为９３－８８（单位：ＩＯＤ值） ，癌周组织端粒酶活性为２４－０９，两者的差别有统计学意义（Ｐ＜ ０－００１） 。正常组织（ｎ ＝ ５） 中未检测到端粒酶活性。端粒酶活性与ＡＦＰ浓度的相关系数ｒ＝０ ．７２，与肿瘤大小的相关系数ｒ＝ －０－２９８。人肝癌高转移模型裸鼠ＬＣＩＤ２０ 的端粒酶活性为（７７－６７±６－９３），低转移模型裸鼠ＬＣＩＤ３５ 的为８２－１６ ±１５－９３），两者之间并无显著性差异（ Ｐ＞ ０－０５） 。 结论 肝癌组织的端粒酶活性明显地高于癌周组织，正常组织中未检测到端粒酶活性。端粒酶活性与ＡＦＰ及肿瘤大小无关。

PURPOSE The experimernts are desired to detect the telomerase activity in hepatocarcinoma,and to understand the relationship between telomerase activity and AFP level, metastatic ability.METHODS Telomerase extract was prepared from the tissues by using TRAP methods,followed by reverse transcription,PCR amplification,dot blot with digtoxigenin labeled probe and silver staining.RESULTS The telomerase activity of hepatoma tissues (n=11) from HCC patients was determeined.It was found that strong telomerase activity was detected in hapatoma tissue,and weak activity in tumorsurrounding tissues.The average telomerase activity in hepatoma tissues was 93.88 IOD,and that in tumor surrounding tissues was 24.09 IOD. The telomerase activity in the tumor tissues both from experimental nude mice model of human HCC with high metastatic ability(LCI D20) was 77.67 IOD and that in low metastaticability (LCI D35) was 82.16 IOD.CONCLUSIONS The telomerase activity in tumor tissues was sifnificantly higher than that in the tissues surrounding tumors ( P <0 001).There is no direct relationship between the telomerase activity and AFP level(r=0.72),tumor size(r=-0.298).The significant difference was not found between these two groups with high and low metastatic ability.It may indicate that the telomease activity was not related to the metastatic potential of HCC.