摘要目的 探讨聚合酶Ⅰ和转录物释放因子(PTRF)在非小细胞肺癌中的功能.方法 使用干扰RNA质粒构建的方法构建2条短发卡RNA(shRNA)s和1条对照shRNA,构建成功后进行空白组、对照组、2组实验组(A组、B组)的转染,使用反转录-聚合酶链反应(RT-RCR)进行目的PTRF的基因水平检测,同时使用蛋白印迹的方法(Western blot)进行蛋白表达水平的检测.然后对A549细胞株进行划痕实验和细胞侵袭实验(Transwells),比较沉默PTRF后对细胞株迁移和侵袭的能力的影响.结果 癌旁组织的PTRF的mRNA和蛋白表达明显高于癌组织,差异有统计学有意义(P=0.003、0.016).shRNA-A组和shRNA-B组PTRF的mRNA相对表达水平显著低于对照组,差异有统计学意义(P =0.009、0.012).shRNA-A组和shRNA-B组PTRF蛋白的相对表达水平显著低于对照组,差异有统计学意义(P =0.002、0.005).转染A549后,使用划痕实验和Transwell侵袭实验进行比较,shRNA-A组和shRNA-B组的细胞迁移和侵袭能力均出现明显的增强,差异有统计学意义(P =0.018、0.029和P=0.024、0.038).结论 非小细胞肺癌通过抑制PTRF的表达促进迁移侵袭.

abstractsObjective To explore the functions of PTRF in non-small cell lung cancer.Methods Two short hairpin RNA (shRNA) and one control shRNA were constructed.And then blank,control,shRNA-A and shRNA-B were transfected in A549 and mRNA and protein levels were detected using reverse transcriptase-polymerase chain reaction (RT-PCR) and Western blotting.Wound healing and invasion assays were done to detect the capability of migration and invasion in A549 cell lines.Results mRNA and protein levels of PTRF in cancer tissues were significantly lower than those in adjacent cancer tissues(P =0.003,P =0.016).mRNA levels of PTRF in shRNA-A and shRNA-B were lower than those in controls(P =0.009,P =0.012).Protein levels of PTRF in shRNA-A and shRNA-B were lower than those in controls(P =0.002,P =0.005).Higher levels of migration and invasion were observed in A549 cell lines after transfection with shRNA-A and shRNA-B (P =0.018,P =0.029 and P =0.024,P =0.038).Conclusion PTRF was decreased and promoted migration in lung cancer.