Institute of Dental Research, Nanjing Medical University, Nanjing, Jiangsu 210029, PR China.
Mol Med Rep. 2012 Feb;5(2):420-6. doi: 10.3892/mmr.2011.668. Epub 2011 Nov 7.
In most types of human cancer, inactivation of pRb/E2F complexes occurs, and released E2Fs initiate transcription of genes required for cell cycle progression. Evidence reveals that phosphorylated pRb deregulates E2F-1, and the levels of E2F-1 expression can accurately predict prognosis of oral squamous cell carcinoma (OSCC). Paradoxically, numerous reports indicate that E2F-1 is also capable of inducing apoptosis under certain cellular circumstances. In the present study, lentivirus-mediated shRNA was used to downregulate endogenous E2F-1 expression in order to study the function of E2F-1 in the pRb/E2F-1 pathway in the OSCC cell line Tca8113, and to investigate the alteration of Tca8113 cells in proliferation and apoptosis. The data from real-time quantitative RT-PCR and Western blot analysis showed that E2F-1-shRNA led to the inhibition of endogenous E2F-1 mRNA and protein expression, and E2F-1 may be associated with proliferation and apoptosis pathways. Growth kinetics data showed that Tca8113-E2F-1-shRNA cells presented more active proliferation properties than Tca8113-NC cells, and flow cytometry data demonstrated that the percentages of cells in the G1 phase, G2 phase and undergoing apoptosis differed between groups. In conclusion, silencing of E2F-1 inhibits proliferation and induces apoptosis. E2F-1 may also be involved in multi-level regulation networks; therefore, its role in OSCC requires further clarification.
在大多数类型的人类癌症中,pRb/E2F 复合物的失活发生,并且释放的 E2F 启动细胞周期进程所需基因的转录。有证据表明,磷酸化的 pRb 使 E2F-1 去调节,E2F-1 的表达水平可以准确预测口腔鳞状细胞癌 (OSCC) 的预后。矛盾的是,许多报道表明,E2F-1 在某些细胞情况下也能够诱导细胞凋亡。在本研究中,使用慢病毒介导的 shRNA 下调内源性 E2F-1 表达,以研究 E2F-1 在 OSCC 细胞系 Tca8113 中的 pRb/E2F-1 通路中的功能,并研究 Tca8113 细胞在增殖和凋亡中的变化。实时定量 RT-PCR 和 Western blot 分析的数据表明,E2F-1-shRNA 导致内源性 E2F-1 mRNA 和蛋白表达的抑制,E2F-1 可能与增殖和凋亡途径有关。生长动力学数据表明,Tca8113-E2F-1-shRNA 细胞比 Tca8113-NC 细胞具有更活跃的增殖特性,而流式细胞术数据表明,各组细胞在 G1 期、G2 期和凋亡期的百分比不同。总之,沉默 E2F-1 抑制增殖并诱导细胞凋亡。E2F-1 可能还参与多层次调节网络;因此,其在 OSCC 中的作用需要进一步阐明。