Zhou Ziliang, Zhou Lijie, Jiang Fangfang, Zeng Binghui, Wei Changbo, Zhao Wei, Yu Dongsheng
Guanghua School of Stomatology, Guangdong Provincial Key Laboratory of Stomatology, Sun Yat-sen University, Guangzhou, P.R. China.
Oncol Res. 2017 Jan 26;25(2):207-214. doi: 10.3727/096504016X14732772150460.
Previous studies have shown that miR-222 targets the p53 upregulated modulator of apoptosis (PUMA) to regulate cell biological behavior in some human malignancies. We hypothesized that there was a negative regulation, which might induce apoptosis, between miR-222 and PUMA in adenoid cystic carcinoma (ACC). In this study, the expression levels of miR-222 and the PUMA gene after transfection with antisense miR-222 (As-miR-222) were evaluated by RT-PCR and Western blot assays. Cell proliferation and migratory abilities were detected by CCK-8 and Transwell assays. Cell cycle and apoptosis were analyzed by flow cytometry. Our results showed that, when compared with the control and scramble-transfected groups, the expression of miR-222 in the As-miR-222 group was downregulated, while the expression of PUMA at both mRNA and protein levels was upregulated, cell proliferation and migratory abilities were inhibited, and apoptosis was increased. Our results suggested that As-miR-222 transfection could upregulate the expression of PUMA to induce apoptosis in ACC, providing a new concept for the treatment of ACC.
先前的研究表明,在一些人类恶性肿瘤中,miR-222靶向p53上调的凋亡调节因子(PUMA)来调控细胞生物学行为。我们推测在腺样囊性癌(ACC)中,miR-222与PUMA之间存在可能诱导细胞凋亡的负调控关系。在本研究中,通过RT-PCR和蛋白质免疫印迹分析评估了用反义miR-222(As-miR-222)转染后miR-222和PUMA基因的表达水平。通过CCK-8和Transwell分析检测细胞增殖和迁移能力。通过流式细胞术分析细胞周期和细胞凋亡。我们的结果显示,与对照组和乱序转染组相比,As-miR-222组中miR-222的表达下调,而PUMA在mRNA和蛋白质水平的表达均上调,细胞增殖和迁移能力受到抑制,细胞凋亡增加。我们的结果表明,As-miR-222转染可上调PUMA的表达以诱导ACC细胞凋亡,为ACC的治疗提供了新的思路。