研究重组TFAR19(TF-1 cell apoptsis-related gene 19)蛋白对鼻咽癌细胞株CNE-1端粒酶活性及端粒酶逆转录酶（hTERT）mRNA表达的影响，探讨重组TFAR19蛋白促CNE-1细胞凋亡的作用机制。方法：将不同浓度的高纯度重组TFAR19蛋白分别和人类鼻咽癌细胞株CNE-1共同培养后，通过7-AAD及Annexin V-PE标记进行流式细胞术分析，检测TFAR19蛋白对细胞的促凋亡效应；RT-PCR法比较试验组和对照组CNE-1细胞株端粒酶hTERT mRNA的表达水平；TRAP-银染法检测重组TFAR19蛋白作用下CNE-1细胞株端粒酶活性的改变。结果：(1) 一定浓度的TFAR19蛋白在未加载体的情况下，可促进细胞凋亡，加入 5、10、15、20、30 mg/L的TFAR19蛋白后, CNE-1细胞的凋亡率分别是:15.26％，29.48％，3711％，5420％，72.36％。阳性对照组（凋亡诱导剂stuanrosporin 处理）与阴性对照组（PBS处理）的细胞凋亡率分别是9837％、1340％。(2) 试验组与对照组CNE-1细胞株端粒酶hTERT mRNA表达无显著差异。(3) 与20 mg/L以上浓度的重组TFAR19蛋白共同培养后，CNE-1细胞株端粒酶活性明显降低。结论：重组TFAR19蛋白在较高浓度下可降低鼻咽癌细胞株CNE-1的端粒酶活性，明显促进肿瘤细胞凋亡。

To investigate the effect of rTFAR19 protein on the expression of hTERT mRNA and telomerase activity in human nasopharyngeal carcinoma CNE-1 cells, so as to understand the mechanism by which rTFAR19 protein promotes the apoptosis of CNE-1 cells. Methods: Different concentrations of rTFAR19 protein were co-cultured with human CNE-1 cells. The apoptosis of CNE-1 cells was analyzed by 7-AAD and Annexin V-PE-labeled flow cytometry (FACS) assay. The expression of hTERT mRNA was detected by RT-PCR and was compared between the 2 groups. The changes of telomerase activity after TFAR19 treatment were examined by TRAP-sliver staining. Results: rTFAR19 protein incorporated into human nasopharyngeal carcinoma CNE-1 cells without a carrier. When the concentrations of rTFAR19 protein were 5mg/L,10 mg/L, 15 mg/L，20 mg/L, and 30 mg/L, the apoptosis rates of CNE-1 cells were 15.26％，2948％，37.11％，54.20％，and 72.36％, respecitively. The apoptosis rate of the positive control group (treated with stuanrosporin) and the negative control group (treated with PBS) were 98.37% and 13.40％, respectively. The expression of hTERT mRNA in CNE-1 cells was similar between the experimental group and the control group. The telomerase activity of CNE-1 cells was obviously decreased after cultured with rTFAR19 protein (>20 mg/L). Conclusion: High concentration of rTFAR19 protein can depress the telomerase activity of human nasopharyngeal carcinoma CNE-1 cells and subsequently accelerate the apoptosis of CNE-1 cells .

湖南省自然科学基金(No.05JJ40043)