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目的探讨含PTEN基因的重组腺病毒载体(Ad-PTEN)对肺腺癌细胞株A549的体外抑制作用。方法采用细胞内同源重组方法构建PTEN基因重组腺病毒载体,293细胞扩增病毒,纯化后转染至人肺腺癌A549细胞株。观察Ad-PTEN的细胞生长抑制作用、细胞凋亡及对细胞超微结构的影响。结果 RT-PCR鉴定证实重组腺病毒载体Ad-PTEN构建成功。体外转染后能在A549细胞内转录和表达,并诱导A549细胞凋亡,抑制其生长和增殖。结论成功构建了含PTEN基因的重组腺病毒载体Ad-PTEN。Ad-PTEN引起肺腺癌细胞株A549明显的细胞形态学改变,生长增殖抑制,诱导A549细胞凋亡,是PTEN基因抑制肺腺癌细胞增殖的主要作用机制之一。
Objective To investigate the inhibitory effect of PTEN gene-containing recombinant adenovirus vector (Ad-PTEN) on lung adenocarcinoma cell line A549 in vitro. Methods Recombinant adenovirus vector of PTEN gene was constructed by homologous recombination in human cells. 293 cells were expanded and then transfected into human lung adenocarcinoma A549 cell line. The effect of Ad-PTEN on cell growth inhibition, apoptosis and cell ultrastructure was observed. Results RT-PCR analysis confirmed that recombinant adenovirus Ad-PTEN was successfully constructed. After transfected in vitro, it can be transcribed and expressed in A549 cells and induce apoptosis of A549 cells, inhibiting its growth and proliferation. Conclusion The recombinant adenovirus Ad-PTEN containing PTEN gene was successfully constructed. Ad-PTEN induced lung adenocarcinoma cell line A549 obvious morphological changes, inhibition of growth and proliferation, apoptosis induced A549 cells, PTEN gene is one of the main mechanisms of inhibition of lung adenocarcinoma cell proliferation.