| NLRP3炎性小体在胃癌细胞中的表达及对其焦亡、增殖、侵袭、迁移的影响 |
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| 引用本文： | 侯滨芬,赵莉,邓敏. NLRP3炎性小体在胃癌细胞中的表达及对其焦亡、增殖、侵袭、迁移的影响[J]. 蚌埠医学院学报, 2022, 47(5): 570-575. DOI: 10.13898/j.cnki.issn.1000-2200.2022.05.003 |
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| 作者姓名： | 侯滨芬 赵莉 邓敏 |
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| 作者单位： | 蚌埠医学院第一附属医院 消化内科，安徽 蚌埠 233004 |
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| 基金项目： | 安徽省科技攻关项目(20194a07020022)；;安徽省自然科学研究项目(2008085MN282)； |
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| 摘 要： | 目的 研究胃癌细胞中NLRP3炎性小体的表达及其对增殖、侵袭和迁移的影响。并初步探讨NLRP3介导caspase-1依赖的焦亡信号通路在胃癌发展中的作用。 方法qRT-PCR和Western blotting法分别检测人胃黏膜上皮细胞(GES-1)和人胃癌细胞(MKN45及MGC803)中NLRP3、caspase-1表达；随后下调NLRP3，检测胃癌细胞NLRP3、caspase-1 mRNA和蛋白的表达；Western blotting检测GSDMD、白细胞介素(IL)-18、IL-1β蛋白表达；ELISA法检测细胞培养液上清液中IL-1β、IL-18含量；乳酸脱氢酶(LDH)实验、流式细胞术检测细胞焦亡；CCK-8法及细胞周期试剂盒检测细胞增殖情况；Transwell和划痕实验检测胃癌细胞侵袭和迁移能力。 结果NLRP3、caspase-1在胃癌细胞中较GES-1表达增高(P < 0.01)。下调NLRP3后，胃癌细胞LDH释放率及焦亡率显著降低(P < 0.01)；GSDMD、IL-1β、IL-18表达降低(P < 0.01)；细胞增殖阻滞在G1期(P < 0.01)；细胞增殖以及侵袭迁移能力显著减弱。 结论NLRP3在胃癌中高表达，其表达影响胃癌细胞的增殖、侵袭和迁移以及其介导的caspase-1依赖焦亡信号通路关键蛋白的表达。
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| 关 键 词： | 胃肿瘤 NLRP3炎性小体 细胞焦亡 caspase-1 |
| 收稿时间： | 2021-10-19 |
Expression of NLRP3 inflammasome in gastric cancer cells and its effect on the pyrophosis,proliferation,invasion and migration |
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| HOU Bin-fen,ZHAO Li,DENG Min. Expression of NLRP3 inflammasome in gastric cancer cells and its effect on the pyrophosis,proliferation,invasion and migration[J]. Journal of Bengbu Medical College, 2022, 47(5): 570-575. DOI: 10.13898/j.cnki.issn.1000-2200.2022.05.003 |
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| Authors: | HOU Bin-fen ZHAO Li DENG Min |
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| Affiliation: | Department of Gastroenterology, The First Affiliated Hospital of Bengbu Medical College, Bengbu Anhui 233030, China |
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| Abstract: | ObjectiveTo investigate the expression of NLRP3 inflammasome in gastric cancer(GC) and its effects on the proliferation, invasion and migration of cells, and the role of NLRP3-mediated caspase-1-dependent pyroptosis signaling pathway in the development of GC. MethodsThe expression levels of NLRP3 and caspase-1 in human gastric epithelial cells(GES-1) and human gastric cancer cells(MKN45 and MGC803) were detected by qRT-PCR and Western blotting.After the expression level of NLRP3 was down-regulated, and the mRNA and protein expression levels of NLRP3 and caspase-1 were detected, the protein expression levels of GSDMD, IL-18 and IL-1β were detected using Western blotting.The content of IL-1β and IL-18 in supernatant were detected by ELISA, the pyroptosis was detected by LDH and flow cytometry, and the cell proliferation was detected by CCK-8 assay and cell cycle kit.Transwell and wound healing were used to detect the invasion and migration of cells. ResultsThe expression levels of NLRP3 and caspase-1 in GC cells were higher than those in GES-1(P < 0.01).After the expression level of NLRP3 was downregulated, the release LDH rate and pyroptosis rate of LDH significantly decreased(P < 0.01), and the expression levels of GSDMD, IL-1β and IL-18 decreased(P < 0.01).Compared with the NC group, the cell proliferation was arrested in G1 phase(P < 0.01).The proliferation, invasion and migration of siNLRP3 cells were significantly reduced. ConclusionsThe expression of NLRP3 in GC cells is high, and its expression affects the proliferation, invasion and migration of GC cells and expression of key proteins in caspase-1-dependent pyroptosis signaling pathway mediated by NLRP3. |
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