慢病毒介导 shRNA 沉默 QKI －6基因对人肺腺癌 A549细胞生物学行为的影响

摘要

Objective:To investigate the effects of lentivirus mediated shRNA silencing QKI -6 gene on the mR-NA and protein expression,cell cycle,colony formation and migration in human lung adenocarcinoma A549 cell line. Methods:The QKI -6 -shRNA lentivirus expression vector was constructed and transfected into A549 cells.The cells infected with shRNA with scramble sequence was used as negative control group and A549 cells was used as control group.The expression of QKI -6 mRNA and protein was detected by RT -PCR and Western blot respective-ly.The cell cycle was detected by flow cytometry.The colony formation of different cells was detected by soft agar col-ony formation assay and migration was detected by Transwell assay.Results:The expression of QKI -6 mRNA and protein was down -regulated by QKI -6 silencing.The flow cytometry showed that the cell cycle of the cells transfect-ed by QKI -6 -shRNA vector was blocked at G0 /G1 phase and the cell population in S phase was increased com-pared with the negative control group and control group.The colony formation capability of the cells transfected by QKI-6 -shRNA vector increased significantly compared with the negative control group and control group.The transwell assay showed that the cell migration potentials of the cells transfected by QKI -6 -shRNA vector increased signifi-cantly compared with the negative control group and control group.Conclusion:QKI -6 could suppress the prolifera-tion,colony formation and migration of the lung adenocarcinoma cell.The QKI -6 gene is a promising anti -tumor target of lung cancer.%目的：探讨慢病毒介导 shRNA 沉默 QKI －6基因对人肺腺癌 A549细胞 QKI －6基因表达、细胞周期、克隆形成和细胞迁移的影响。方法：构建 QKI －6－shRNA 慢病毒表达载体，并用其转染 A549细胞，阴性对照组转染无意义序列，空白对照组为 A549细胞。运用 RT －PCR 及 Western blot 检测各组细胞 QKI －6 mR-NA 及蛋白的表达，流式细胞仪检测细胞周期，琼脂成瘤实验检测细胞克隆形成能力，Transwell 实验检测细胞迁移能力。结果：实验组 QKI －6 mRNA 及蛋白表达均较阴性对照组及空白对照组降低。实验组较阴性对照组及空白对照组，G0／G1期细胞比例降低（P ＜0．05），S 期细胞比例增高（P ＜0．05）。琼脂成瘤实验显示，实验组与空白对照组及阴性对照组比较，克隆形成率明显提高（P ＜0．05）。细胞迁移实验结果显示，实验组与空白对照组及阴性对照组比较，细胞迁移数量明显增多（P ＜0．05）。结论：QKI －6能够抑制肺腺癌细胞生长、克隆形成及迁移，QKI －6有望成为肺癌治疗的新靶点。