Zhou Kai, Fan Yan-Dong, Duysenbi Serick, Wu Peng-Fei, Feng Zhao-Hai, Qian Zheng, Zhang Ting-Rong
Department of Neurosurgery, The First Affiliated Hospital of Xinjiang Medical University, Urumqi, P.R. China.
Tumour Biol. 2017 Jun;39(6):1010428317704805. doi: 10.1177/1010428317704805.
Human pituitary adenoma is one of the most common intracranial tumors with an incidence as high as 16.7%. Recent evidence has hinted a relationship between growth factors of pituitary or hypothalamic origin and proliferation of human pituitary adenoma cells. This study explores the effects of small interfering RNA-mediated silencing of basic fibroblast growth factor gene on the proliferation, migration, and invasion of human pituitary adenoma cells. Human pituitary adenoma tissues were collected to obtain human pituitary adenoma cells. The basic fibroblast growth factor silencing interference plasmid was constructed, and the human pituitary adenoma cells were transfected and assigned as basic fibroblast growth factor-small interfering RNA, negative control-small interfering RNA, and blank groups. The quantitative real-time polymerase chain reaction and Western blotting were carried out to detect the expression of basic fibroblast growth factor, pituitary tumor transforming gene, vascular endothelial growth factor, cluster of differentiation 147, and matrix metalloproteinase 9. Cell Counting Kit-8 assay was conducted to observe cell proliferation at 0, 24, 48, and 72 h. Flow cytometry was used to determine cell cycle. Transwell and scratch test were applied to detect the invasion and migration of pituitary adenoma cells. Protein kinase C activity was detected. In comparison with the blank group, the basic fibroblast growth factor-small interfering RNA group showed reduced messenger RNA and protein expression of basic fibroblast growth factor, reduced cell viability at 24, 48, and 72 h, increased cells in G0/G1 stage, declined cells in S and G2/M stages, decreased number of cell migration, shortened migrating distance, reduced protein kinase C activity, and decreased expression of pituitary tumor transforming gene, vascular endothelial growth factor, cluster of differentiation 147, and matrix metalloproteinase 9. However, the negative control-small interfering RNA group had no evident differences in basic fibroblast growth factor expression, cell viability, cell cycle, number of cell migration, migrating distance, protein kinase C activity, and expression of pituitary tumor transforming gene, vascular endothelial growth factor, cluster of differentiation 147, and matrix metalloproteinase 9 compared with the blank group. The study provides evidence that small interfering RNA-mediated silencing of basic fibroblast growth factor gene inhibits the proliferation, migration, and invasion of human pituitary adenoma cells.
垂体腺瘤是最常见的颅内肿瘤之一,发病率高达16.7%。近期证据提示,垂体或下丘脑来源的生长因子与垂体腺瘤细胞增殖之间存在关联。本研究探讨小干扰RNA介导的碱性成纤维细胞生长因子基因沉默对垂体腺瘤细胞增殖、迁移及侵袭的影响。收集垂体腺瘤组织以获取垂体腺瘤细胞。构建碱性成纤维细胞生长因子沉默干扰质粒,转染垂体腺瘤细胞并分为碱性成纤维细胞生长因子-小干扰RNA组、阴性对照-小干扰RNA组和空白组。采用定量实时聚合酶链反应和蛋白质印迹法检测碱性成纤维细胞生长因子、垂体肿瘤转化基因、血管内皮生长因子、分化簇147和基质金属蛋白酶9的表达。采用细胞计数试剂盒-8法观察0、24、48和72小时的细胞增殖情况。采用流式细胞术检测细胞周期。采用Transwell和划痕试验检测垂体腺瘤细胞的侵袭和迁移能力。检测蛋白激酶C活性。与空白组相比,碱性成纤维细胞生长因子-小干扰RNA组碱性成纤维细胞生长因子的信使核糖核酸和蛋白表达降低,24、48和72小时细胞活力降低,G0/G1期细胞增多,S期和G2/M期细胞减少,细胞迁移数量减少,迁移距离缩短,蛋白激酶C活性降低,垂体肿瘤转化基因、血管内皮生长因子、分化簇147和基质金属蛋白酶9的表达降低。然而,阴性对照-小干扰RNA组与空白组相比,碱性成纤维细胞生长因子表达、细胞活力、细胞周期、细胞迁移数量、迁移距离、蛋白激酶C活性以及垂体肿瘤转化基因、血管内皮生长因子、分化簇147和基质金属蛋白酶9的表达均无明显差异。该研究证实,小干扰RNA介导的碱性成纤维细胞生长因子基因沉默可抑制垂体腺瘤细胞的增殖、迁移及侵袭。