作者：张欢,石文健,刘永亮,李景武,张于,董桂兰,董伟

单位：

1.063000 河北省唐山市人民医院神经外科 2.063000 河北省唐山市人民医院肿瘤外科　 3.063000 河北省唐山市人民医院重症医 学科 4.063000 河北省唐山市人民医院肿瘤内科 通信作者：董伟，E-mail：polly0522@yeah.net

Units：

1.Department of Neurosurgery, Tangshan People's Hospital, Tangshan 063000, China 2.Department of Tumor Surgery, Tangshan People's Hospital, Tangshan 063000, China 3.Department of Intensive Care Medicine, Tangshan People's Hospital, Tangshan 063000, China 4.Department of Oncology, Tangshan People's Hospital, Tangshan 063000, China Corresponding author: DONG Wei, E-mail: polly0522@yeah.net

关键词：

垂体肿瘤; 低氧; 细胞增殖; 细胞侵袭; 上皮-间质转化;

Keywords：

Pituitary neoplasms; Hypoxia; Cell proliferation; Cell invasion; Epithelial-mesenchymal transition

CLC：

DOI：

10.12114/j.issn.1008-5971.2021.00.268

Funds：

河北省自然科学基金精准医学联合基金细化项目（H2020105017）

摘要：

背景 约35%的垂体腺瘤呈侵袭性生长,而上皮-间质转化(EMT)与肿瘤的侵袭和复发密切相关。低氧会在多种肿瘤中诱导EMT,但其在垂体腺瘤中的作用尚不清楚。目的 探究低氧诱导大鼠垂体腺瘤GH3细胞增殖、侵袭和EMT的机制。方法 本实验时间为2020年9月至2021年6月。将GH3细胞分为常氧组〔不转染低氧诱导因子(HIF)-1α-siRNA,氧浓度为19.5%〕、低氧组(不转染HIF-1α-siRNA,氧浓度为2.5%)、常氧+siRNA组(转染HIF-1α-siRNA,氧浓度为19.5%)、低氧+siRNA组(转染HIF-1α-siRNA,氧浓度为2.5%)。比较各组干扰后24、48、72 h细胞增殖能力,细胞侵袭能力,GH3细胞中HIF-1α、N-钙黏蛋白(N-Cad)、波形蛋白(Vim)mRNA及其蛋白表达水平。结果 低氧组干扰后24、48、72 h细胞增殖能力高于常氧组(P <0.05);常氧+siRNA组干扰后72 h细胞增殖能力低于常氧组,干扰后24、48、72 h细胞增殖能力低于低氧组(P <0.05);低氧+siRNA组干扰后24 h细胞增殖能力高于常氧组,干扰后24、48、72 h细胞增殖能力低于低氧组、高于常氧+siRNA组(P<0.05)。常氧组、低氧组、常氧+siRNA组、低氧+siRNA组干扰后48、72 h细胞增殖能力分别高于本组干扰后24 h(P <0.05);常氧组、低氧组、常氧+siRNA组、低氧+siRNA组干扰后72 h细胞增殖能力分别高于本组干扰后48 h(P <0.05)。低氧组细胞侵袭能力高于常氧组(P <0.05);常氧+siRNA组细胞侵袭能力低于常氧组、低氧组(P <0.05);低氧+siRNA组细胞侵袭能力低于低氧组,高于常氧+siRNA组(P <0.05)。低氧组GH3细胞中HIF-1α、N-Cad、Vim mRNA表达水平高于常氧组(P <0.05);常氧+siRNA组GH3细胞中HIF-1α、Vim mRNA表达水平低于常氧组,HIF-1α、N-Cad、Vim mRNA表达水平低于低氧组(P <0.05);低氧+siRNA组GH3细胞中HIF-1α、N-Cad、Vim mRNA表达水平低于低氧组,HIF-1α、Vim mRNA表达水平高于常氧+siRNA组(P <0.05)。低氧组GH3细胞中HIF-1α、N-Cad、Vim蛋白表达水平高于常氧组(P <0.05);常氧+siRNA组GH3细胞中HIF-1α、N-Cad、Vim蛋白表达水平低于常氧组、低氧组(P <0.05);低氧+siRNA组GH3细胞中Vim蛋白表达水平高于常氧组,HIF-1α、N-Cad、Vim蛋白表达水平低于低氧组、高于常氧+siRNA组(P <0.05)。结论 低氧通过上调HIF-1α表达而促进GH3细胞增殖和细胞侵袭,低氧通过上调N-Cad、Vim表达而促进EMT的发生,而干扰HIF-1α表达可部分阻断这些作用。

Abstract：

【Abstract】　Background About 35% of pituitary adenomas grow aggressively, and epithelial-mesenchymal transition(EMT) is closely related to tumor invasion and recurrence. Hypoxia can induce EMT in a variety of tumors, but its role in pituitaryadenomas is unclear. ObjectiveTo explore the mechanism of cell proliferation, cell invasion and EMT of rat pituitary adenomaGH3 cells induced by hypoxia.MethodsThe study period was from September 2020 to June 2021. GH3 cells were divided intonormoxia group [without transfection of hypoxic inducible factor (HIF)-1α-siRNA, oxygen concentration was 19.5%] , hypoxiagroup (without transfection of HIF-1α-siRNA, oxygen concentration was 2.5%) , normoxia+siRNA group (transfected with HIF-1α-siRNA, oxygen concentration was 19.5%) , hypoxia+siRNA group (transfected with HIF-1α-siRNA, oxygen concentrationwas 2.5%) . The cell proliferation ability at 24, 48, and 72 h after the interference, cell invasion ability, HIF-1α, N-cadherin(N-Cad) , vimentin (Vim) mRNA and protein expression levels in GH3 cells were compared in each group.ResultsThecell proliferation ability of the hypoxia group was higher than that of the normoxia group at 24, 48, and 72 h after interference(P < 0.05) . The cell proliferation ability of the normoxia+siRNA group was lower than that of the normoxia group at 72 h after theinterference, and the cell proliferation ability was lower than that of the hypoxia group at 24, 48, and 72 h after the interference(P < 0.05) . The cell proliferation ability of the hypoxia+siRNA group was higher than that of the normoxia group at 24 h afterinterference, and the cell proliferation ability was lower than that of the hypoxia group and higher than that of the normoxia+siRNAgroup at 24, 48, and 72 h after the interference (P < 0.05) . In the normoxia group, hypoxia group, normoxia+siRNA group, andhypoxia+siRNA group, the cell proliferation ability at 48 and 72 h after interference was higher than that at 24 h after interference,respectively (P < 0.05) . In the normoxia group, hypoxia group, normoxia+siRNA group, and hypoxia+siRNA group, the cellproliferation capacity at 72 h after interference was higher than that at 48 h after interference, respectively (P < 0.05) . The cellinvasion ability of hypoxia group was higher than that of normoxia group (P < 0.05) . The cell invasion ability of normoxia+siRNAgroup was lower than that of normoxia group and hypoxia group (P < 0.05) . The cell invasion ability of hypoxia+siRNA groupwas lower than that of hypoxia group and higher than that of normoxia+siRNA group (P < 0.05) . The expression levels of HIF-1α, N-Cad and Vim mRNA in GH3 cells in the hypoxia group were higher than those in the normoxia group (P < 0.05) . Theexpression levels of HIF-1α and Vim mRNA in GH3 cells in the normoxia+siRNA group were lower than those in the normoxiagroup, and the expression levels of HIF-1α, N-Cad and Vim mRNA were lower than those in the hypoxia group (P < 0.05) .The expression levels of HIF-1α, N-Cad and Vim mRNA in GH3 cells in the hypoxia+siRNA group were lower than those inthe hypoxia group, and the expression levels of HIF-1α and Vim mRNA were higher than those in the normoxia+siRNA group(P < 0.05) . The expression levels of HIF-1α, N-Cad and Vim protein in GH3 cells in the hypoxia group were higher thanthose in the normoxia group (P < 0.05) . The expression levels of HIF-1α, N-Cad and Vim protein in GH3 cells in thenormoxia+siRNA group were lower than those in the normoxia group and hypoxia group (P < 0.05) . The expression level ofVim protein in GH3 cells in the hypoxia+siRNA group was higher than that in the normoxia group, and the expression levels ofHIF-1α, N-Cad and Vim protein were lower than that in the hypoxia group and higher than that in the normoxia+siRNA group(P < 0.05) .Conclusion Hypoxia promotes GH3 cell proliferation and cell invasion by up-regulating the expression level ofHIF-1α, hypoxia promotes the occurrence of EMT by up-regulating the expression levels of N-Cad and Vim, while interferencewith the expression of HIF-1α can partially block these effects.

ReferenceList：