SDF-1α/miR-134 轴调控非功能性垂体神经内分泌肿瘤生长并靶向 VEGFA。

SDF-1α/MicroRNA-134 Axis Regulates Nonfunctioning Pituitary Neuroendocrine Tumor Growth Targeting VEGFA.

机构信息

Department of Neurosurgery, The Second Affiliated Hospital, School of Medicine, Zhejiang University, Hangzhou, Zhejiang, China.

Department of Surgical Oncology, The Second Affiliated Hospital, School of Medicine, Zhejiang University, Hangzhou, Zhejiang, China.

出版信息

Front Endocrinol (Lausanne). 2020 Dec 9;11:566761. doi: 10.3389/fendo.2020.566761. eCollection 2020.

DOI:10.3389/fendo.2020.566761

PMID:33362712

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC7756115/

Abstract

BACKGROUND

Nonfunctioning pituitary neuroendocrine tumor (NF-PitNET) is difficult to resect. Except for surgery, there is no effective treatment for NF-PitNET. MicroRNA-134 (miR-134) has been reported to inhibit proliferation and invasion ability of tumor cells. Herein, the mechanism underlying the effect of miR-134 on alleviating NF-PitNET tumor cells growth is explored.

METHODS

Mouse pituitary αT3-1 cells were transfected with miR-134 mimics and inhibitor, followed by treatment with stromal cell-derived factor-1α (SDF-1α) . MiR-134 expression level: we used quantitative real-time PCR (qRT-PCR) to detect the expression of miR-134. Cell behavior level: cell viability and invasion ability were assessed using a cell counting kit-8 (CCK8) assay and Transwell invasion assay respectively. Cytomolecular level: tumor cell proliferation was evaluated by Ki-67 staining; propidium iodide (PI) staining analyzed the effect of miR-134 on cell cycle arrest; western blot analysis and immunofluorescence staining evaluated tumor migration and invasive ability. Additionally, we collected 27 NF-PitNET tumor specimens and related clinical data. The specimens were subjected to qRT-PCR to obtain the relative miR-134 expression level of each specimen; linear regression analysis was used to analyze the miR-134 expression level in tumor specimens and the age of the NF-PitNET population, gender, tumor invasion, prognosis, and other indicators.

RESULTS

experiment, miR-134 was observed to significantly inhibit αT3-1 cells proliferation characterized by inhibited cell viability and expressions of vascular endothelial growth factor A (VEGFA) and cell cycle transition from G1 to S phase ( < 0.01). VEGFA was verified as a target of miR-134. Additionally, miR-134-induced inhibition of αT3-1 cell proliferation and invasion was attenuated by SDF-1α and VEGFA overexpression ( < 0.01). In primary NF-PitNET tumor analysis, miR-134 expression level was negatively correlated with tumor invasion ().

CONCLUSION

The regulation of the SDF-1α/miR-134/VEGFA axis represents a novel mechanism in the pathogenesis of NF-PitNETs and may serve as a potential therapeutic target for the treatment of NF-PitNETs.

摘要

背景

无功能性垂体神经内分泌肿瘤（NF-PitNET）难以切除。除手术外，NF-PitNET 尚无有效治疗方法。MicroRNA-134（miR-134）已被报道可抑制肿瘤细胞的增殖和侵袭能力。在此，我们探讨了 miR-134 缓解 NF-PitNET 肿瘤细胞生长的作用机制。

方法

用 miR-134 模拟物和抑制剂转染鼠垂体αT3-1 细胞，然后用基质细胞衍生因子-1α（SDF-1α）处理。miR-134 表达水平：我们使用实时定量 PCR（qRT-PCR）检测 miR-134 的表达。细胞行为水平：通过细胞计数试剂盒-8（CCK8）检测和 Transwell 侵袭检测分别评估细胞活力和侵袭能力。细胞分子水平：通过 Ki-67 染色评估肿瘤细胞增殖；碘化丙啶（PI）染色分析 miR-134 对细胞周期阻滞的影响；Western blot 分析和免疫荧光染色评估肿瘤迁移和侵袭能力。此外，我们收集了 27 例 NF-PitNET 肿瘤标本和相关临床资料。对标本进行 qRT-PCR 以获得每个标本的相对 miR-134 表达水平；线性回归分析用于分析肿瘤标本中的 miR-134 表达水平与 NF-PitNET 人群的年龄、性别、肿瘤侵袭、预后等指标的关系。

结果

实验中，miR-134 明显抑制 αT3-1 细胞增殖，表现为细胞活力降低，血管内皮生长因子 A（VEGFA）表达降低，细胞从 G1 期向 S 期过渡（<0.01）。VEGFA 被验证为 miR-134 的靶标。此外，SDF-1α 和 VEGFA 过表达可减弱 miR-134 诱导的 αT3-1 细胞增殖和侵袭抑制作用（<0.01）。在原发性 NF-PitNET 肿瘤分析中，miR-134 表达水平与肿瘤侵袭呈负相关（）。