食管上皮癌变过程中端粒长度、hTERT含量的定量研究

Telomere length and hTERT expression in carcinogenesis of esophageal epithelium

目的:研究端粒DNA长度、hTERT蛋白定量表达与食管上皮细胞癌变的关系。方法:对84例食管上皮脱落细胞分别应用流式荧光原位杂交检测端粒DNA长度,细胞免疫荧光流式定量法检测hTERT蛋白含量。结果:①正常食管上皮脱落细胞组、重度增生组和癌组增殖指数(PI)分别为11.51±4.08,19.04±5.39和29.46±5.50,癌组细胞增殖活性显著高于重度增生组(P<0.01),重度增生组高于正常组(P<0.01),PI值与细胞学分级呈正相关(r=0.80,P<0.01)。②正常食管上皮脱落细胞组、重度增生组和癌组的端粒长度Q-FISH值分别为50.83±8.86、36.96±8.02和27.81±6.59,癌组端粒长度明显短于重度增生组(P<0.01),重度增生组短于正常组(P<0.01)。端粒长度与细胞学分级呈负相关(r=-0.73,P<0.01)。③正常组、重度增生组和癌组hTERT蛋白荧光指数(FI)分别为0.95±0.14、1.47±0.22和1.75±0.19,癌组hTERT蛋白含量高于重度增生组(P<0.01),重度增生组高于正常组(P<0.01)。hTERT含量与细胞学分级呈正相关(r=0.81,P<0.01)。重度增生组和癌组hTERT蛋白阳性表达率分别为91.43%(32/35)和96.77%(30/31),二者都明显高于正常组(P<0.01)。结论:端粒DNA长度缩短、hTERT蛋白含量升高与食管上皮癌变密切相关。

Objective： To investigate the correlations of telomere length and expression level of human telomerase reverse transcriptase （hTERT） protein to the carcinogenesis of esophageal epithelium. Methods：Telomere length was measured by quantitative flow cytometry based fluorescence in situ hybridization （Q-Flow FISH） and hTERT protein expression was examined by flow cytometry with indirect immunofluorescence staining in 84 cases of desquamated esophageal epithelial cells. Results：①The value of proliferation index （Pl） was 11.51±4.08,19.04±5.39,and 29.46±5.50 in normal group,severe dysplasia group, and carcinoma group, respectively. The Pl value in carcinoma group was higher than that in severe dysplasia group（P〈0.01）. The P1 value in severe dysplasia group was higher than that of normal group（P〈0.01）. The Pl value had a positive correlation with cytological grade（r=0.80,P〈0.01）. ② The value of Q-FISH indicating telomere length was 50.83±8.86, 36.96±8.02, and 27.81± 6. 59 in normal group, severe dysplasia group, and carcinoma group, respectively. The telomere length in carcinoma group was markedly shorter than that in severe dysplasia group（P〈0.01）. The telomere length of severe dysplasia group was shorter than that in normal group（ P〈0.01 ）. The telomere length had a negative correlation with cytological grade （r= -0.73, P〈0.01）. ③Value of fluorescence index （Fl） indicating expression level of hTERT protein was 0. 95±0.14,1. 47±0. 22,and 1. 75±0. 19 in normal group,severe dysplasia group, and carcinoma group, respectively. The expression level of hTERT protein in carcinoma group was higher than that in severe dysplasia group（P〈0.01）. The expression level of hTERT protein in severe dysplasia group was higher than that in normal group（P〈0.01）. The FI value had a positive correlation with the cytological grade （r= 0.81, P〈0.01 ）. hTERT protein was positive in 32 of the 35 dysplasia specimens （91.43 %） and in 30 of the 31 carcinoma specimens（96.77% ）. The positive ratio of hTERT protein in dysplasia and carcinoma group were higher than that in normal group（P〈0.01 ）. Conelusion： The telomere shortening and hTERT protein over-expression had a close correlation with carcinogenesis of esophageal epithelium.