目的 探究过表达低氧诱导因子-1α (hypoxia inducible factor 1α, HIF-1α)对人脑胶质瘤SHG44细胞恶性度的影响及其可能机制。方法 体外培养SHG44细胞,转染过表达HIF-1α入SHG44细胞,评估HIF-1α转染效率;对比分析HIF-1α过表达对SHG44细胞的增殖、干性、迁移和侵袭的影响;ELISA法检测肿瘤相关炎性反应因子(TNF-α,IL-10,IL-17和IL-1β),Western blot检测SHG44细胞焦亡相关蛋白[炎性小体3(inflammasome3,NLRP3)、凋亡相关斑点样蛋白(apoptosis-associated speck-like protein,ASC)、天冬氨酸蛋白水解酶1(cysteinyl aspartate specific proteinase-1,caspase-1)、GSDMD、GSDME和转化生长因子-β1(transforming growth factor-β1,TGF-β1)]的表达,明确HIF-1α对焦亡的关系。结果 HIF-1α过表达后SHG44细胞明显肿胀增多,细胞的增殖、干性、迁移和侵袭能力较对照组明显增强,与对照组相比肿瘤炎性因子(TNFα,IL-10和IL-1β)表达水平均明显升高,而肿瘤炎性反应抑制因子IL-17表达水平明显下降;HIF-1α过表达后焦亡相关蛋白(NLRP3、ASC、caspase-1、GSDMD、GSDME和TGF-β1)表达明显上调(P<0.05)。结论 HIF-1α过表达可能通过促进焦亡提高SHG44细胞的增殖、干性、迁移和侵袭能力。
Abstract
Objective To investigate the effect of hypoxia inducible factor-1 α (HIF-1α) overexpression on the malignancy of human glioma SHG44 cells and its possible mechanism.Methods Glioma cell SHG44 was cultured in vitro and transfected with HIF-1α overexpression into SHG44 cells. The effects of HIF-1α overexpression on the growth and proliferation, cell stem, invasion and metastasis of tumor cells in vitro were analyzed. Finally, the relationship between HIF-1α and pyroptosis were clarified by detecting the expression of HIF-1α, tumor-associated inflammatory factors(TNF- α, IL-10, IL-17 and IL-1 β), pyroptosis markers [inflammatory body 3 (NLRP3), apoptosis associated speck like protein (ASC), cysteinyl aspartate specific proteinase-1 (caspase-1), GSDMD, GSDME and transforming growth factor-β1(TGF-β1nd).Results MTT, cell cloning, cell scratch and Transwell experiments confirmed that the morphological swelling of SHG44 cells increased significantly after the overexpression of HIF-1α, and cell proliferation and invasion ability were significantly enhanced compared with the control group; ELISA showed that, compared with the control group, tumor inflammatory factor (TNF α,IL-10 and IL-1 β)increased significantly, while IL-17 decreased significantly; RT-PCR and Western blot showed that the expression level of HIF-1α in glioma cell line was higher than that in normal brain cells The expression levels of NLRP3, ASC, caspase-1, GSDMD and GSDME inHIF-1αoverexpression group were significantly higher than those in control group (P<0.05).Conclusions The increased expression of HIF-1α in glioma cells SHG44 can increase the proliferation and invasion of glioma cells, which may be achieved by promoting pyroptosis.
关键词
低氧诱导因子-1α /
神经胶质细胞 /
SHG44细胞 /
焦亡 /
转染
Key words
hypoxia inducible factor 1α /
glial cells /
SHG44 cells /
pyroptosis /
transfection
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基金
武警后勤部科研项目(CWJ18L004);武警后勤学院基础研究项目(WHJ201721);福建省自然科学基金(2020J011273)
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