反义端粒酶RNA对肺癌细胞A549抑制作用研究

目的 探讨反义人端粒酶RNA(human telomeric,RNA,hTR)对肺癌细胞A549端粒酶活性和细胞增殖的抑制作用，研究以端粒酶为靶的肺癌基因治疗的可能性。方法 将端粒酶hTR的cDNA反向插入到逆转录病毒载体pLXRN中，转染包装细胞PT67后获得反义hTR重组病毒，感染肺癌细胞A549。处理前后采用TRAP法检测端粒酶活性，光镜观察细胞形态，DNA电泳观察凋亡情况。结果 作用后端粒酶活性和细胞增殖受到明显抑制，并出现明显的细胞凋亡。结论 反义hTR 对肺癌细胞A549端粒酶活性具有明显的抑制作用，使肺癌细胞A549生长抑制，促进细胞凋亡。

THE INHIBITION OF ANTISENSE hTR ON A549 LUNG CANCER CELLS

ObjectiveTo investigate the inhibition of telomerase activity and cell growth in lung cancer cells A549 by antisense hTR (human telomeric RNA) treatment, and explore the possibility of the telomerase as a target for lung cancer gene therapy.MethodsThe antisense hTR fragment was inserted into the pLXRN plasmid, obtain the replication-deficient retroviruses by transfection the packing cells of PT67, then infect the A549 cells by the antisense virus. Test telomerase activity of the treated and untreated cells by using telomeric repeat amplification protocol (TRAP), observe the cells morphological change by microscope, and assay the apoptosis by DNA electrophoresis.ResultsThe cells growth and telomerase activity of A549 were significantly inhibited and apoptosis occurred after antisense virus infection when compared with the control cell lines. ConclusionThe introduction of antisense telomeric RNA can significantly inhibit the activity of telomerase, arrest the cells growth and accelerate apoptosis in A549.