SYBR Green染色法检测肺癌及癌旁组织端粒酶活性

目的 研究TRAP－SYBR Green染色法定性检测端粒酶活性的方法及端粒酶活性在肺癌及癌旁组织中的表达，并探讨端粒酶的表达与P53之间可能的关系。方法 采用TRAP－SYBR Green染色法检测36例肺癌、36例癌旁组织、8例肺部良性病变组织和A549肺腺癌细胞株端粒酶活性的表达，并利用免疫组化技术检测肺癌组织P53蛋白表达水平。结果 TRAP－SYBR Green染色法可检测出5个A549肺腺癌细胞所表达端粒酶活性。83．3％（30／36）的肺癌组织，3／8肺部良性病变组织，19．4％（7／36）的癌旁组织表达端粒酶活性。端粒酶活性的表达与肺癌组织的病理类型、肿瘤大小、临床分期无相关性。在30例表达端粒酶活性的肺癌组织中P53呈现过度表达的占80％（24／30），端粒酶活性与P53蛋白表达水平之间具有相关关系。结论 TRAP－SYBR Green染色法是一种较为灵敏、快速、简便的定性检测端粒酶活性的方法；端粒酶活性的定性检测可作为肺部恶性肿瘤诊断的辅助指标之一；端粒酶P53之间的关系有待进一步阐明。

Telomerase activity in lung cancer and adjacent peritumoral tissues determined by TRAP-SYBR green assay

Objective To develop methods for the qualitative analysis of telomerase activity by TRAP-SYBR Green assay and study telomerase activity in lung cancer and adjacent peritumoral tissues, and to explore the possible association with expression of P53. Methods Thirty six lung cancer tissues and 36 adjacent peritumoral tissues, 8 lung benign lesion tissues and A549 lung adenoma cell line, were examined for telomerase activity by TRAP-SYBR Green assay. The 36 lung cancer tissue samples were also examined for P53 protein expression by immunohistochemical staining. Results Telomerase activity in 5 cells of A549 lung cancer cell line was detected by TRAP-SYBR Green assay. Thirty of 36 (83.3%) lung cancer specimens, seven of 36 (19.4%) adjacent peritumoral tissues, three of 8 (37.5%) lung benign lesion tissues were positive for telomerase activity. Telomerase activity of lung cancer did not correlate with pathologic type, tumor size and clinical stage. P53 was over-expressed in 24 of the 30 (80%) lung cancer samples with telomerase activity. A significant correlation was found between telomerase activity and accumulation of P53 proteins (r=0.60, P<0.01).Conclusion TRAP-SYBR Green assay may be a sensitive, fast and convenient method for qualitative analysis of telomerase activity, which is useful in diagnosis lung neoplasms. The relationship between telomerase activity and P53 accumulation needs further study.