Zhang Kai-Liang, Zhou Xuan, Han Lei, Chen Lu-Yue, Chen Ling-Chao, Shi Zhen-Dong, Yang Ming, Ren Yu, Yang Jing-Xuan, Frank Thomas S, Zhang Chuan-Bao, Zhang Jun-Xia, Pu Pei-Yu, Zhang Jian-Ning, Jiang Tao, Wagner Eric J, Li Min, Kang Chun-Sheng
Department of Neurosurgery, Tianjin Medical University General Hospital; Laboratory of Neuro-Oncology, Tianjin Neurological Institute; Key Laboratory of Post-trauma Neuro-repair and Regeneration in Central Nervous System, Ministry of Education; Tianjin Key Laboratory of Injuries, Variations and Regeneration of Nervous System, Tianjin 300052, China.
Mol Cancer. 2014 Mar 20;13:63. doi: 10.1186/1476-4598-13-63.
Epidermal growth factor receptor (EGFR) is amplified in 40% of human glioblastomas. However, most glioblastoma patients respond poorly to anti-EGFR therapy. MicroRNAs can function as either oncogenes or tumor suppressor genes, and have been shown to play an important role in cancer cell proliferation, invasion and apoptosis. Whether microRNAs can impact the therapeutic effects of EGFR inhibitors in glioblastoma is unknown.
miR-566 expression levels were detected in glioma cell lines, using real-time quantitative RT-PCR (qRT-PCR). Luciferase reporter assays and Western blots were used to validate VHL as a direct target gene of miR-566. Cell proliferation, invasion, cell cycle distribution and apoptosis were also examined to confirm whether miR-566 inhibition could sensitize anti-EGFR therapy.
In this study, we demonstrated that miR-566 is up-regulated in human glioma cell lines and inhibition of miR-566 decreased the activity of the EGFR pathway. Lentiviral mediated inhibition of miR-566 in glioblastoma cell lines significantly inhibited cell proliferation and invasion and led to cell cycle arrest in the G0/G1 phase. In addition, we identified von Hippel-Lindau (VHL) as a novel functional target of miR-566. VHL regulates the formation of the β-catenin/hypoxia-inducible factors-1α complex under miR-566 regulation.
miR-566 activated EGFR signaling and its inhibition sensitized glioblastoma cells to anti-EGFR therapy.
在40%的人类胶质母细胞瘤中，表皮生长因子受体（EGFR）会发生扩增。然而，大多数胶质母细胞瘤患者对抗EGFR治疗反应不佳。微小RNA可作为癌基因或抑癌基因发挥作用，并且已被证明在癌细胞增殖、侵袭和凋亡中起重要作用。微小RNA是否会影响EGFR抑制剂对胶质母细胞瘤的治疗效果尚不清楚。
使用实时定量逆转录聚合酶链反应（qRT-PCR）检测胶质瘤细胞系中miR-566的表达水平。荧光素酶报告基因检测和蛋白质免疫印迹法用于验证VHL是miR-566的直接靶基因。还检测了细胞增殖、侵袭、细胞周期分布和凋亡情况，以确认抑制miR-566是否能使抗EGFR治疗更敏感。
在本研究中，我们证明miR-566在人类胶质瘤细胞系中上调，抑制miR-566可降低EGFR通路的活性。慢病毒介导的对胶质母细胞瘤细胞系中miR-566的抑制显著抑制了细胞增殖和侵袭，并导致细胞周期停滞在G0/G1期。此外，我们确定了冯·希佩尔-林道（VHL）是miR-566的一个新的功能靶点。VHL在miR-566的调控下调节β-连环蛋白/缺氧诱导因子-1α复合物的形成。
miR-566激活EGFR信号，抑制它可使胶质母细胞瘤细胞对抗EGFR治疗更敏感。
