Liu Xiaowen, Sun Jun, Yuan Ping, Shou Kangquan, Zhou Yuanhong, Gao Wenqi, She Jin, Hu Jun, Yang Jun, Yang Jian
1Hubei Key Laboratory of Tumor Microenvironment and Immunotherapy, Medical College, China Three Gorges University, Yichang, China.
2Institute of Cardiovascular Research & Department of Center Experiment Laboratory, the First College of Clinical Medical Science, China Three Gorges University, Yichang, 44300 China.
Cancer Cell Int. 2019 Jul 29;19:197. doi: 10.1186/s12935-019-0916-9. eCollection 2019.
Mitofusin 2 (Mfn2) is outer membrane protein, as the inhibitor of Ras protein. This study aimed to investigate the effect of Mfn2 on cell proliferation, and cell-cycle in Hela cervical carcinoma cell lines.
After treated with Adv-mfn2 or Adv-control for 48 h and 60 h, the RNA and protein of Mfn2 in Hela cells were detected by qRT-PCR and western blot. The immunofluorescence assay was performed to observe the expression and sub-location of Mfn2 in Hela cells. The flow cytometry was performed to detect the cell cycle of Hela cells, while western blots were performed to observe the Ras-NF-κB signal pathway. Then, the xenografted cervix carcinoma mouse model was used to confirm the effect of Mfn2 in Hela cells in vivo and the expression of Ras-NF-κB signaling pathway in vivo.
In immunofluorescence detection, Mfn2 was located in cytoplasmic, not in the nucleus. In addition, Mfn2 inhibited cell proliferation of Hela cells through reducing PCNA protein expression. Mfn2 induced arrest in G0/G1 phase of the cell cycle in Hela cells. Meanwhile, Mfn2 reduced Cyclin D1 protein expression. Moreover, Mfn2 decreased the Ras signal pathway proteins such as Myc, NF-κB p65, STAT3 in a dose-dependent manner. Then, the in vivo experiment also confirmed that Mfn2 could inhibit the tumor growth, and depress the Cyclin D1, Ras, Myc, NF-κB p65, Erk1/2 and mTOR protein expression.
Mfn2 could significantly inhibit cell proliferation in Hela cells. It might be acted as an potential anti-cancer target through inducing cell cycle arrest in human cervical carcinoma cells.
线粒体融合蛋白2（Mfn2）是一种外膜蛋白，作为Ras蛋白的抑制剂。本研究旨在探讨Mfn2对人宫颈癌Hela细胞系细胞增殖及细胞周期的影响。
用Adv-mfn2或Adv-control处理Hela细胞48小时和60小时后，采用qRT-PCR和蛋白质免疫印迹法检测Hela细胞中Mfn2的RNA和蛋白质。进行免疫荧光分析以观察Mfn2在Hela细胞中的表达及亚定位。采用流式细胞术检测Hela细胞的细胞周期，同时用蛋白质免疫印迹法观察Ras-NF-κB信号通路。然后，利用宫颈癌异种移植小鼠模型在体内证实Mfn2对Hela细胞的作用以及Ras-NF-κB信号通路在体内的表达。
在免疫荧光检测中，Mfn2定位于细胞质，而非细胞核。此外，Mfn2通过降低增殖细胞核抗原（PCNA）蛋白表达抑制Hela细胞的增殖。Mfn2诱导Hela细胞的细胞周期停滞于G0/G1期。同时，Mfn2降低细胞周期蛋白D1（Cyclin D1）的蛋白表达。而且，Mfn2以剂量依赖的方式降低Ras信号通路蛋白如Myc、NF-κB p65、信号转导和转录激活因子3（STAT3）的表达。然后，体内实验也证实Mfn2可抑制肿瘤生长，并降低Cyclin D1、Ras、Myc、NF-κB p65、细胞外信号调节激酶1/2（Erk1/2）和雷帕霉素靶蛋白（mTOR）的蛋白表达。
Mfn2可显著抑制Hela细胞的增殖。它可能通过诱导人宫颈癌细胞的细胞周期停滞而成为一种潜在的抗癌靶点。
