Department of Neurosurgery, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, P.R. China.
J Cell Biochem. 2018 Apr;119(4):3162-3173. doi: 10.1002/jcb.26469. Epub 2017 Dec 26.
This current study intends to investigate the effect of microRNA-128 (miR-128) on cisplatin (DDP) resistance in glioma SHG-44 cells. SHG-44/DDP cells were transfected with miR-128 antisense oligonucleotide (ASO) and assigned into blank, resistance, NC, anti-miR-128, miR-128 mimic, si-JAG1, and anti-miR-128 + si-JAG1 groups. qRT-PCR and Western blotting were employed for determining expression of miR-128, JAG1, Bax and Bcl-2. MTT assay, Giemsa staining, and flow cytometry were applied to detect DDP resistance, cellular morphology, and cell cycle, respectively. JAG1 is targeted and negatively regulated by miR-128. In in vitro experiments, compared with the blank group, the rest groups exhibited declined miR-28 and Bax expression, lowered cell inhibition rate and apoptosis rate, but elevated JAG1 and Bcl-2 expression with cells arrested in the S phase. Compared with the resistance group, the anti-miR-128 group showed decreasedBax expression along with a lowered cell inhibition rate and apoptosis rate, but increased JAG1 and Bcl-2 expression with reduced cells arrested in the S phase; while the miR-128 mimic group showed an opposite trend; the si-JAG1 group showed decreased Bcl-2 expression and reduced cells in the S phase. In in vivo experiments, compared with the resistance group, the tumor growth rate, tumor volume, and weight as well as JAG1 expression accelerated in the anti-miR-128 group; whereas the miR-128 mimic and si-JAG1 groups exhibited an opposite trend. Our findings demonstrated that miR-128 ASO transfection might down-regulate the expression of miR-128 in SHG-44/DDP and up-regulate the DDP resistance in SHG-44/DDP cells, providing a potential treatment target for glioma.
本研究旨在探讨微小 RNA-128（miR-128）对脑胶质瘤 SHG-44 细胞顺铂（DDP）耐药性的影响。将 miR-128 反义寡核苷酸（ASO）转染 SHG-44/DDP 细胞，分为空白组、耐药组、NC 组、抗 miR-128 组、miR-128 模拟物组、si-JAG1 组和抗 miR-128+si-JAG1 组。采用 qRT-PCR 和 Western blot 检测 miR-128、JAG1、Bax 和 Bcl-2 的表达。MTT 法、吉姆萨染色法和流式细胞术分别用于检测 DDP 耐药性、细胞形态和细胞周期。JAG1 是 miR-128 的靶基因，并受其负调控。在体外实验中，与空白组相比，其余各组 miR-28 和 Bax 表达降低，细胞抑制率和凋亡率降低，而 JAG1 和 Bcl-2 表达升高，S 期细胞阻滞。与耐药组相比，抗 miR-128 组 Bax 表达降低，细胞抑制率和凋亡率降低，而 JAG1 和 Bcl-2 表达升高，S 期细胞减少；miR-128 模拟物组则呈现相反的趋势；si-JAG1 组 Bcl-2 表达降低，S 期细胞减少。在体内实验中，与耐药组相比，抗 miR-128 组肿瘤生长速度、肿瘤体积和重量以及 JAG1 表达加快；而 miR-128 模拟物和 si-JAG1 组则呈现相反的趋势。本研究结果表明，miR-128 ASO 转染可能下调 SHG-44/DDP 中 miR-128 的表达，上调 SHG-44/DDP 细胞的 DDP 耐药性，为脑胶质瘤的治疗提供了一个潜在的靶点。
