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目的 :探讨端粒长度、端粒酶活性以及端粒酶亚单位组分 (hTERT、hTR)的表达与食管上皮细胞永生化和恶性转化之间的关系。方法 :对HPV18E6E7基因诱发的人胎儿食管上皮永生化细胞系SHEE和恶性转化细胞系SHEEC ,通过Southernblot检测端粒长度 (TRF) ,TRAP法测定端粒酶活性 ,RT PCR研究端粒酶催化亚单位 (hTERT)和端粒酶RNA成分 (hTR)的表达。结果 :SHEE细胞和SHEEC细胞的端粒平均长度比正常食管上皮细胞的明显缩短 ,但稳定维持在一定长度范围内。SHEE细胞和SHEEC细胞均具有端粒酶活性 ,并均有hTERT和hTR表达。结论 :端粒酶表达活化使端粒维持在一定长度是永生化食管上皮细胞SHEE和恶性转化细胞SHEEC能够稳定分裂增殖的重要因素之一。
Objective: To investigate the relationship between telomere length, telomerase activity, expression of telomerase subunits (hTERT, hTR) and immortalization and malignant transformation of esophageal epithelial cells. METHODS: Human fetal esophageal epithelial immortalized cell line SHEE and malignant transformed cell line SHEEC were induced by HPV18E6E7 gene. Telomere length (TRF) was detected by Southern blot, telomerase activity was measured by TRAP assay, and telomerase catalytic subunit was studied by RT PCR. (hTERT) and telomerase RNA component (hTR) expression. Results: The average telomere length of SHEE cells and SHEEC cells was significantly shorter than that of normal esophageal epithelial cells, but remained stable within a certain length. SHEE cells and SHEEC cells all have telomerase activity, and all have hTERT and hTR expression. Conclusion : Activation of telomerase expression to maintain telomere at a certain length is one of the important factors for the stable division and proliferation of immortalized esophageal epithelial cell SHEE and malignant transformed cell SHEEC.