目的 探究环指蛋白187(ring finger protein 187,RNF187)过表达后对人脑胶质瘤细胞U87的影响及其可能机制。方法 体外培养胶质瘤细胞系U87,转染过表达RNF187入U87细胞,对比分析RNF187 过表达对肿瘤细胞生长增殖能力、细胞干性和体外侵袭及转移能力;最后通过检测RNF187及焦亡标志物,即炎性小体3(inflammasome3,NLRP3)、凋亡相关斑点样蛋白(apoptosis-associated speck-like protein,ASC)、GSDMD和GSDME的表达,明确RNF187与焦亡的关系。结果 CCK-8、细胞克隆和Transwell 实验证实,RNF187过表达后U87细胞形态上肿胀肿大,且细胞增殖和侵袭能力较对照组增强,RNF187过表达组细胞增殖率(2.24±0.16)高于其他两组;RT-PCR和Western blot实验表明,RNF187过表达后焦亡相关标志物(NLRP3、ASC、GSDMD、GSDME)表达水平较对照组升高(P＜0.05)。结论 RNF187在胶质瘤细胞表达增多,增加胶质瘤细胞的增殖和侵袭能力,可能是通过促进焦亡实现的。
Abstract
Objective To investigate the effect of RNF187 overexpression on human glioma cell U87 and the possible mechanism. Methods Glioma cell U87 was cultured in vitro and transfected with overexpressed RNF187 into U87 cells. The effects of RNF187 overexpression on the growth, proliferation, cell stemness, invasion and metastasis of tumor cells in vitro were analyzed. Finally, the relationship between RNF187 and pyroptosis was studied by detecting the expressions of RNF187 and markers of pyroptosis[inflammatory body 3 (NLRP3), apoptosis associated speck like protein (ASC), GSDMD and GSDME]. Results CCK-8, cell cloning and Transwell experiments confirmed that the morphological swelling of U87 cells increased significantly after overexpression of RNF187, and the cells' ability of proliferation and invasion was significantly enhanced compared with the control group. RT-PCR and Western blot showed that the expression level of RNF187 in glioma cell line was higher than that in normal brain cells. The expression levels of NLRP3, ASC, caspase-1, GSDMD and GSDME in RNF187 the overexpression group were significantly higher than those of the control group (P＜0.05). Conclusions The higher expression of RNF187 in glioma cells can increase the proliferation and invasion of glioma cells, possibly by promoting pyroptosis.
关键词
神经胶质瘤细胞 /
环指蛋白187 /
焦亡 /
恶性度
Key words
glioma cells /
ring finger protein 187 /
pyroptosis /
malignancy
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基金
福建自然科学基金(2020J011273), 武警后勤学院基础研究项目(WHJ201721)
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