Department of Pharmacy, Jiangxi Provincial People's Hospital, Nanchang, Jiangxi 330006, P.R. China.
Department of General Surgery, Jiangxi Provincial People's Hospital, Nanchang, Jiangxi 330006, P.R. China.
Mol Med Rep. 2018 Jun;17(6):8053-8060. doi: 10.3892/mmr.2018.8921. Epub 2018 Apr 23.
The present study aimed to investigate the anticancer effect of sorafenib combined with silencing of activating transcription factor 2 (ATF2) in hepatocellular carcinoma (HCC) cells and to assess the underlying molecular mechanisms. Huh‑7 HCC cell line was selected for the present study. Small interfering RNA (siRNA)‑ATF2 sequence was constructed to silence ATF2 expression. The experiment was divided into 6 groups: i) Control; ii) vector; iii) sorafenib (6.8 µM); iv) vector+sorafenib; v) siRNA‑ATF2; and vi) siRNA‑ATF2+sorafenib groups. Cell proliferation, apoptosis, migration and invasion were detected following treatments with sorafenib and/or ATF2 silencing. Additionally, expression of tumor necrosis factor (TNF)‑α and c‑Jun N‑terminal kinase 3 (JNK3) was detected using reverse transcription‑quantitative polymerase chain reaction and western blotting. The current findings revealed that siRNA‑ATF2 significantly reduced ATF2 expression. Cell proliferation, migration and invasion abilities in the sorafenib and siRNA‑ATF2 groups were significantly reduced compared with the control group (P<0.05). Apoptotic rate in the sorafenib and siRNA‑ATF2 groups was significantly increased compared with the control group (P<0.05). The mRNA and protein expression levels of ATF2 in the sorafenib or siRNA‑ATF2 groups was significantly reduced when compared with control group. The phosphorylation of ATF2 was also reduced following sorafenib treatment or ATF2 silence. Although JNK3 mRNA expression level was not affected, the phosphorylation level of JNK3 was significantly promoted following sorafenib treatment or ATF2 silencing. Additionally, TNF‑α mRNA and protein expression levels were increased following sorafenib treatment or ATF2 silencing. It is of note that siRNA‑ATF2 treatment promoted the anticancer activity of sorafenib in Huh‑7 cells. Additionally, siRNA‑ATF2+sorafenib treatment combined additionally promoted TNF‑α expression and phosphorylation of JNK3. Combined siRNA‑ATF2 and sorafenib treatment had a greater anticancer effect compared with sorafenib or ATF2 silencing alone. The possible mechanism involved in the anticancer effect of sorafenib and ATF2 silencing may be associated with the activation of the TNF‑α/JNK3 signaling pathway.
本研究旨在探讨索拉非尼联合沉默激活转录因子 2（ATF2）对肝癌（HCC）细胞的抗癌作用，并评估其潜在的分子机制。选择 Huh-7 HCC 细胞系进行本研究。构建了小干扰 RNA（siRNA）-ATF2 序列以沉默 ATF2 表达。实验分为 6 组：i）对照组；ii）载体组；iii）索拉非尼（6.8μM）组；iv）载体+索拉非尼组；v）siRNA-ATF2 组；和 vi）siRNA-ATF2+索拉非尼组。用索拉非尼和/或 ATF2 沉默处理后，检测细胞增殖、凋亡、迁移和侵袭。此外，采用逆转录-定量聚合酶链反应和蛋白质印迹法检测肿瘤坏死因子（TNF）-α 和 c-Jun N-末端激酶 3（JNK3）的表达。目前的研究结果表明，siRNA-ATF2 可显著降低 ATF2 表达。与对照组相比，索拉非尼和 siRNA-ATF2 组的细胞增殖、迁移和侵袭能力显著降低（P<0.05）。与对照组相比，索拉非尼和 siRNA-ATF2 组的凋亡率显著增加（P<0.05）。与对照组相比，索拉非尼或 siRNA-ATF2 组的 ATF2 mRNA 和蛋白表达水平显著降低。索拉非尼处理或 ATF2 沉默后，ATF2 磷酸化水平也降低。尽管 JNK3 mRNA 表达水平不受影响，但索拉非尼处理或 ATF2 沉默后 JNK3 的磷酸化水平显著升高。此外，索拉非尼处理或 ATF2 沉默后 TNF-α mRNA 和蛋白表达水平增加。值得注意的是，siRNA-ATF2 处理可增强索拉非尼在 Huh-7 细胞中的抗癌活性。此外，siRNA-ATF2+索拉非尼治疗联合促进 TNF-α 表达和 JNK3 磷酸化。与单独使用索拉非尼或 ATF2 沉默相比，联合使用 siRNA-ATF2 和索拉非尼治疗具有更强的抗癌作用。索拉非尼和 ATF2 沉默的抗癌作用的可能机制与 TNF-α/JNK3 信号通路的激活有关。
