Title: [XAF1 inhibits cell proliferation and induces apoptosis in human lung adenocarcinoma cell line A549 in vitro].
Abstract: XAF1 is a factor necessary to inhibit tumor cell growth. Low XAF1 expression is associated with various tumor cells. The aim of this study is to investigate the effect and the mechanism of adenovirus vector Ad5/F35 mediated X-linked inhibitor of apoptosis protein associated factor-1 (XAF1) on the inhibition of cell proliferation and the induction of apoptosis of human lung adenocarcinoma cell A549.Recombinant virus Ad5/F35-XAF1 and controlled virus Ad5/F35-NULL exhibited different multiplicities of infection (MOI) at the same time. mRNA and protein expressions of XAF1 were determined by reverse transcriptase polymerase chain reaction (RT-PCR) and Western blot, respectively. Cell proliferation was observed by methyl thiazolyl tetrazolium (MTT) assay, and cell apoptosis was analyzed by FACS with Annexin V-FITC/PI double staining. The expressions of apoptosis-associated proteins, such as PARP, Caspase-3, and Caspase-8, were also determined by Western blot.mRNA and protein expressions of XAF1 were significantly increased in human lung adenocarcinoma cell A549 after this cell was transfected with Ad5/F35-XAF1 for 48 h; these expressions were higher than those of the controlled group Ad5/F35-NULL. Cell proliferation was inhibited and apoptosis was induced in a dose-dependent manner in the Ad5/F35-XAF1 group. After Ad5/F35-XAF1 transfection was performed, the cleavage of apoptosis-associated proteins, such as PARP, Caspase-3, and Caspase-8, was activated.Restored XAF1 expression inhibits cell proliferation and induces cell apoptosis in human lung adenocarcinoma cell line A549. Furthermore, XAF1 may activate associated apoptotic signaling pathways in A549 cell line.背景与目的 XAF1是重要的肿瘤细胞生长抑制因子,其低表达与多种肿瘤细胞有关。研究肿瘤抑制基因XAF1对人肺腺癌细胞株A549的作用及机制。方法 利用重组腺病毒Ad5/F35-XAF1和对照腺病毒Ad5/F35-NULL瞬时转染A549细胞,用逆转录聚合酶链式反应(reverse transcriptase polymerase chain reaction, RT-PCR)和Western blot方法检测A549细胞株中XAF1 mRNA和蛋白质的表达;MTT检测细胞增殖率、流式细胞仪检测细胞凋亡率,并用Western blot法检测凋亡相关蛋白的表达。结果 腺病毒介导的XAF1瞬时转染肺腺癌A549细胞后,XAF1 mRNA及蛋白表达水平明显提高,并能明显抑制该细胞增殖和促进细胞凋亡,蛋白质印记法显示凋亡相关蛋白PARP、Caspase-3、Caspase-8的裂解条带。结论 恢复XAF1基因在人肺腺癌A549细胞中表达后,能明显抑制该肿瘤细胞增殖并促进其凋亡,其机制可能与XAF1激活肺癌细胞相关凋亡途径有关。
Publication Year: 2014
Publication Date: 2014-12-01
Language: en
Type: article
Indexed In: ['doaj', 'pubmed']
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Cited By Count: 3
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