Gene silencing of BAG-1 modulates apoptotic genes and sensitizes lung cancer cell lines to cisplatin-induced apoptosis
Gene silencing of BAG-1 modulates apoptotic genes and sensitizes lung cancer cell lines to cisplatin-induced apoptosis
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BAG-1 基因沉默调节凋亡基因并使肺癌细胞系对顺铂诱导的细胞凋亡敏感
DOI：
10.4161/cbt.9.10.11589
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发表时间：
2010-05-15
影响因子：
3.6
通讯作者：
Zhou, Qinghua
中科院分区：
文献类型：
--
作者：
Liu, Hongyu;Liang, Yonggang;Zhou, Qinghua
BAG-1是一种多功能蛋白，可与多种靶分子相互作用，调节细胞凋亡。然而，BAG-1抑制细胞凋亡的确切机制尚不清楚。在本研究中，我们通过比较BAG-1 siRNA沉默后在人肺腺癌细胞A549中的基因表达谱，全面探讨BAG-1介导的基因表达变化。微阵列分析显示589个基因表达改变，包括177个上调基因和412个下调基因。实时荧光定量PCR检测结果显示，基因表达变化与基因芯片检测结果的一致性为86.48%（64/74）。上调的基因包括EGFR、Jun、HSPA1A和casp7；下调的基因包括Lyn、HRAS和TGF α。此外，我们通过western blot证实，BAG-1基因沉默后，EGFR、Jun和HSPA1A的表达显著升高，Lyn和HRAS的表达显著降低。转染BAG-1 siRNA后，肺癌细胞系A549和L9981对顺铂诱导的细胞凋亡的敏感性显著增强。因此，BAG-1基因的沉默导致凋亡相关基因的表达改变，使A549和L9981细胞对顺铂诱导的凋亡敏感。
BAG-1 is a multifunctional protein that interacts with a wide range of target molecules to regulate apoptosis. However, the exact mechanism by which BAG-1 inhibits apoptosis is unknown. In the current study, we comprehensively investigate BAG-1 mediated changes in gene expression by comparing the gene expression profile after BAG-1 siRNA silencing in the human lung adenocarcinoma cell line A549. Microarray assay revealed that expression of 589 genes was altered, including 177 upregulated and 412 downregulated genes. Real-time PCR assay of the changes in gene expression indicated a concordance of 86.48% (64/74) with that observed by microarray. The upregulated genes included EGFR, Jun, HSPA1A and CASP 7; while the downregulated genes included Lyn, HRAS and TGF alpha. Furthermore, we confirmed by western blot that the expression of EGFR, Jun and HSPA1A were significantly increased, while expression of Lyn and HRAS were dramatically decreased after BAG-1 gene silencing. After transfection with BAG-1 siRNA, lung cancer cell lines A549 and L9981 exhibited significantly enhanced sensitivity to cisplatin-induced apoptosis. Thus, silencing of the BAG-1 gene resulted in changes in expression of apoptosis-related genes and sensitized A549 and L9981 cells to cisplatin-induced apoptosis.