腺病毒介导的CIAPIN1基因对肺癌细胞生长和细胞周期的影响

Influence of adenovirusomediated CIAPIN1 gene on cell growth and cell cycle in lung cancer

目的以重组腺病毒载体介导细胞因子诱导的凋亡抑制分子1（cytokine induced apoptosis inhibitor 1，CIAPIN1）基因转染人肺腺癌细胞系A549，观察并鉴定该基因在腺癌细胞中的表达及其对细胞生长和细胞周期的影响。方法构建腺病毒载体介导CIAPIN1基因（Ad—CIAPIN1），转染到低水平表达CIAPIN1的人肺腺癌细胞系A549，采用Western blot检测目的蛋白的表达；台盼蓝染色计数活细胞数，绘制细胞生长曲线；流式细胞术（FCM）分析细胞周期及细胞凋亡的变化。结果外源性CIAPIN1基因在A549肺癌细胞获得高水平的表达；CIAPIN1能显著抑制A549的生长（P〈0．01）；流式细胞仪检测显示肺癌细胞发生凋亡，并出现明显的Gt／s期阻滞现象。结论以腺病毒为载体介导CIAPIN1基因在A549细胞内表达，可发挥抑制细胞生长、诱发细胞凋亡及参与细胞周期调控的作用。提示Ad—CIAPIN1基因可能在人肺癌基因治疗中发挥重要作用。

Objective To investigate the CIAPIN1 gene expression in lung adenocarcinoma cell line A549 transduced by adenovirus（Ad） and observe the effects of exogenous CIAPIN1 gene on cell growth and cell cycle. Methods An adenoviral expression vector with fulllength cDNA of CIAPIN1 gene insert was constructed （Ad-CIAPIN1） and transfected into A549 cells. The expression of CIAPIN1 protein was identified by Western blot. Trypan blue staining was used to count the living cells and to draw a cellular growth curve. The changes of cell cycle and apoptosis were analyzed by flow cytometry（FCM）. Results High expression of exogenous CIAPIN1 gene in A549 cells was confirmed by Western blot. The growth of the Ad-CIAPIN1 transfected A549 cells in vitro was significantly inhibited. Marked alterations in the cell cycle with a predominant arrest in the G1/S phase, followed by a large fraction of apoptotic cells were found by FCM. Conclusions Transduction of A549 CIAPIN1-negative cell with Ad-CIAPIN1 inhibited cell growth and altered cell cycle progression. These results demonstrate that CIAPIN1 plays an important role in the development of lung cancer and strengthens the hypothesis of its possible use as a therapeutic target for lung cancer.