Study on the inhibitory effect of antisense PNA of telomerase for growth of lung cancer cell lines in vitro

Abstract

目的 探讨反义端粒酶肽核酸(PNA)片段对肺癌细胞株端粒酶活性及细胞株生长的抑制作用.方法 将人工合成的端粒酶反义PNA片段,应用脂质体转染法将反义端粒酶序列导入肺腺癌细胞株A549及小细胞癌细胞株NCI-H446中,采用MTT法检测活细胞数,采用RT-PCR-ELLISA法检测端粒酶活性.结果 转染72 h后,A549、NCI-H446细胞株端粒酶活性(A450值)分别由0.582±0.039和0.571±0.043降低至0.294±0.048(P＜0.01)和0.276±0.051(P＜0.01);而两细胞株的活细胞数(A580值)分别由0.485±0.009和0.513±0.015降低至0.191±0.027(P＜0.01)和0.138±0.046(P＜0.01),细胞生长受到显著抑制.结论 反义端粒酶PNA片段具有抑制肺癌细胞株端粒酶活性的作用,并可抑制肺癌细胞的生长.

Objective To determine the inhibitory effect of antisense peptide nucleic acids (PNA) of telomerase on the growth of lung cancer cell lines. Methods The synthesized modified antisense PNAs of telomerase were transfected into the lung cancer cell lines A549 and NCI-H446 respectively by lipofectamine transfection. Telomerase activity was detected by RT-PCR-ELLISA, and the cell counts were determined by MTT. Results Seventy two hours after transfection with antisense PNAs of telomerase, telomerase activity (A450 value) of A549 and NCI-H446 were down regulated from 0.582 ±0.039, 0.571±0.043 to 0.294±0.048 ( P < 0.01), 0.276±0.051 ( P < 0.01) respectively, and alive cell counts (A580 value) of them from 0.485± 0.009 , 0.513±0.015 to 0.191±0.027 ( P < 0.01), 0.138±0.046 ( P < 0.01) respectively. The growth of two lung cancer cell lines were significantly inhibited. Conclusion Antisense PNA of telomerase might inhibit not only the telomerase activity, but also the growth of lung cancer cell lines in vitro .