CT45A1 siRNA silencing suppresses the proliferation, metastasis and invasion of lung cancer cells by downregulating the ERK/CREB signaling pathway.
CT45A1 siRNA silencing suppresses the proliferation, metastasis and invasion of lung cancer cells by downregulating the ERK/CREB signaling pathway.
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DOI：
10.3892/mmr.2017.7466
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发表时间：
2017-11
影响因子：
3.4
通讯作者：
Jia K
中科院分区：
文献类型：
--
作者：
Tang F;Tang S;Guo X;Yang C;Jia K
本研究旨在探讨癌睾丸抗原家族45成员A1（CT45A1）在肺癌细胞增殖、凋亡、侵袭和转移中的作用及其分子机制。 Western blotting检测发现正常肺细胞中CT45A1的表达远低于肺癌细胞中观察到的表达。使用 Lipofectamine 2000 将 CT45A1 小（或短）干扰 (si)RNA 及其阴性对照转染至 A549 细胞后，通过蛋白质印迹和逆转录聚合酶链反应进一步测定 CT45A1 蛋白和 mRNA 水平。转染CT45A1 siRNA后，肺癌细胞中CT45A1水平显着降低（P<0.01）。然后，分别用甲基噻唑基四唑测定和膜联蛋白 V-FITC/碘化丙啶染色研究细胞活力和细胞凋亡。采用Transwell实验评估A549细胞的迁移和侵袭能力。与阴性对照相比，CT45A1 siRNA处理后的肺癌细胞活力、迁移和侵袭均受到抑制，细胞凋亡增加（均P<0.01）。此外，通过蛋白质印迹法评估B细胞淋巴瘤-2 (Bcl-2)、Bcl-2相关X (Bax)、生存素、基质金属蛋白酶2 (MMP2)、MMP9、细胞外信号调节激酶1/2 (ERK1/2)、磷酸化ERK1/2 (p-ERK1/2)、环AMP反应元件结合蛋白(CREB)和p-CREB的水平。 CT45A1沉默后，Bcl-2、survivin、MMP2、MMP9、p-ERK1/2、p-CREB表达下调，Bax表达上调（均P<0.01）。结论 CT45A1 siRNA沉默通过下调ERK/CREB信号通路抑制肺癌细胞的增殖、转移和侵袭。
The aim of the present study was to investigate the role of the cancer-testis antigen family 45 member A1 (CT45A1) in the proliferation, apoptosis, invasion and metastasis of lung cancer cells, and the associated molecular mechanisms. Western blotting determined that the expression of CT45A1 in normal lung cells was far lower than that observed in lung cancer cells. Following the transfection of CT45A1 small (or short) interfering (si)RNA and its negative control into A549 cells using Lipofectamine 2000, the CT45A1 protein and mRNA levels were determined further by western blotting and reverse transcription-polymerase chain reaction. Following CT45A1 siRNA transfection, the levels of CT45A1 in lung cancer cells were markedly reduced (P<0.01). Then, cell viability and apoptosis were investigated with a methyl thiazolyl tetrazolium assay and Annexin V-FITC/propidium iodide staining, respectively. Transwell assays were employed to evaluate the migration and invasion of A549 cells. When compared with the negative control, the viability, migration and invasion of lung cancer cells treated with CT45A1 siRNA were suppressed and apoptosis was promoted (all P<0.01). In addition, the levels of B-cell lymphoma-2 (Bcl-2), Bcl-2 associated X (Bax), survivin, matrix metalloproteinase 2 (MMP2), MMP9, extracellular signal-regulated kinase 1/2 (ERK1/2), phosphorylated ERK1/2 (p-ERK1/2), cyclic AMP response element binding protein (CREB) and p-CREB were assessed by western blotting. Following CT45A1 silencing, the expressions of Bcl-2, survivin, MMP2, MMP9, p-ERK1/2 and p-CREB were downregulated and the expression of Bax was upregulated (all P<0.01). It was concluded that CT45A1 siRNA silencing suppressed the proliferation, metastasis and invasion of lung cancer cells by downregulating the ERK/CREB signalling pathway.
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影响因子：
11.5
作者：
Takezawa, Ken;Okamoto, Isamu;Nakagawa, Kazuhiko
通讯作者：
Nakagawa, Kazuhiko
影响因子：
3.1
作者：
Zhang, De-Hui;Zhang, Liang-Yu;Zhao, Jin-Zao
通讯作者：
Zhao, Jin-Zao
影响因子：
--
作者：
Peng B;Lei N;Chai Y;Chan EK;Zhang JY
通讯作者：
Zhang JY
影响因子：
4
作者：
Wong, Janica C.;Bathina, Madhavi;Fiscus, Ronald R.
通讯作者：
Fiscus, Ronald R.
DOI：
10.1073/pnas.0502583102
发表时间：
2005-05-31
影响因子：
11.1
作者：
Chen, YT;Scanlan, MJ;Simpson, AJG
通讯作者：
Simpson, AJG