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中华老年病研究电子杂志

中华老年病研究电子杂志 ›› 2020, Vol. 07 ›› Issue (04) : 16 -20. doi: 10.3877/cma.j.issn.2095-8757.2020.04.003

所属专题： 文献；

论著

NOD样受体家族蛋白3在中老年子宫内膜样腺癌组织中的表达及意义
周锦红1,†(), 曾淑梅1, 赵灵琴2, 程贤鹦1, 李文君1   
  1. 1. 310013　杭州，浙江医院妇产科
    2. 310022　杭州，浙江省肿瘤医院妇瘤外科
  • 收稿日期:2020-03-24 出版日期:2020-11-28
  • 通信作者: 周锦红

Expression and significance of NOD-like receptor family protein 3 in middle-aged and elderly patients with endometrioid adenocarcinoma

Jinhong Zhou1,†(), Shumei Zeng1, Lingqin Zhao2, Xianying Cheng1, Wenjun Li1   

  1. 1. Department of Obstetrics and Gynecology, Zhejiang Hospital, Hangzhou 310013, China
    2. Women's Oncology Surgery, Zhejiang Cancer Hospital, Hangzhou 310022, China
  • Received:2020-03-24 Published:2020-11-28
  • Corresponding author: Jinhong Zhou
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引用本文：

周锦红, 曾淑梅, 赵灵琴, 程贤鹦, 李文君. NOD样受体家族蛋白3在中老年子宫内膜样腺癌组织中的表达及意义[J]. 中华老年病研究电子杂志, 2020, 07(04): 16-20.

Jinhong Zhou, Shumei Zeng, Lingqin Zhao, Xianying Cheng, Wenjun Li. Expression and significance of NOD-like receptor family protein 3 in middle-aged and elderly patients with endometrioid adenocarcinoma[J]. Chinese Journal of Geriatrics Research(Electronic Edition), 2020, 07(04): 16-20.

目的

探讨NOD样受体家族蛋白3（NOD-like receptor protein, NLRP3）在中老年患者子宫内膜样腺癌组织中的表达及临床意义。

方法

选取2017年1月至2019年12月在浙江医院和浙江省肿瘤医院行手术切除的子宫内膜样腺癌患者35例，以及同期因子宫脱垂行全子宫切除术患者12例。采用实时荧光定量PCR、免疫印迹、免疫组化等方法检测比较不同子宫内膜组织中NLRP3 mRNA和蛋白的表达水平。同时应用子宫内膜样腺癌细胞系ishikawa和HEC-1A行体外细胞试验，观察NLRP3对子宫内膜样腺癌细胞增殖、侵袭及迁移能力的影响。两组间的比较采用t检验。

结果

免疫组化检测：NLRP3在子宫内膜样腺癌组织中的表达水平明显高于正常子宫内膜组织，而且随FIGO分期及病理分级的增高而增高。实时荧光定量PCR检测：子宫内膜样腺癌组织中NLRP3 mRNA和NLRP3蛋白的表达水平均显著高于正常子宫内膜组织（t=3.769、4.316，P<0.05）。蛋白质印迹法：子宫内膜样腺癌组织中NLRP3蛋白表达明显强于正常子宫内膜组织。体外细胞试验：NLRP3 mRNA和NLRP3蛋白在ishikawa细胞和HEC-1A细胞中均有表达，而且两者在HEC-1A细胞中的表达水平均明显高于ishikawa细胞（t=2.373、2.559，P<0.05）；利用siRNA沉默子宫内膜样腺癌ishikawa细胞和HEC-1A细胞的NLRP3后，从第3天开始其增殖能力明显受限，并持续至细胞增殖实验结束的第7天；Transwell实验：沉默NLRP3后ishikawa细胞和HEC-1A细胞的迁移和侵袭数量均明显下降（t=7.343、6.571，3.859、3.289；P<0.05）。

结论

NLRP3的表达与子宫内膜样腺癌细胞的增殖、侵袭和迁移能力密切相关。

关键词: 子宫内膜样腺癌, NOD样受体家族蛋白3, 炎症, 增殖, 迁移, 侵袭
Objective

To investigate the expression and clinical significance of NOD-like receptor protein 3 (NLRP3) in middle-aged and elderly patients with endometrioid adenocarcinoma.

Methods

13 patients with endometrioid adenocarcinoma and underwent surgical resection admitted to Zhejiang Hospital and Zhejiang Cancer Hospital from January 2017 to December 2019 were selected. At the same time, 12 patients with uterine prolapse who underwent total hysterectomy were selected. The expression levels of NLRP3 mRNA and protein were detected by real-time PCR, Western blot and immunohistochemistry. The effects of NLRP3 on proliferation, invasion and migration of endometrial adenocarcinoma cell lines Ishikawa and HEC-1A were observed in vitro. T test was used to compare the measurement data between the groups.

Results

Immunohistochemical analysis showed that the expression level of NLRP3 in endometrioid adenocarcinoma tissues was significantly higher than that in normal endometrium tissues, and increased with the increase of FIGO stage and pathological grade. PCR results showed that the expression levels of NLRP3 mRNA and NLRP3 protein in the endometrioid adenocarcinoma tissue were significantly higher than those in the normal endometrial tissue (t=3.769, 4.316; P < 0.05). Western blot results showed that the expression of NLRP3 protein in endometrioid adenocarcinoma tissues was significantly higher than that in normal endometrium tissues. Cell assay results showed that NLRP3 mRNA and NLRP3 protein were expressed in both Ishikawa cells and HEC-1a cells, and the expression levels of NLRP3 mRNA and NLRP3 protein in HEC-1A cells were significantly higher than those in Ishikawa cells (t =2.373, 2.559; P < 0.05); after silencing NLRP3 of Ishikawa cells and HEC-1A cells with siRNA, the proliferation ability of the cells was significantly limited from the third day, and continued to the seventh day after the end of cell proliferation experiment; transwell assay showed that the number of migration and invasion of Ishikawa cells and HEC-1a cells decreased significantly after NLRP3 silencing (t=7.343, 6.571, 3.859, 3.289; P < 0.05).

Conclusion

The expression of NLRP3 is closely related to the pathology, proliferation, invasion and migration of endometrioid adenocarcinoma.

Key words: Endometrioid adenocarcinoma, NOD-like receptor protein 3, Inflammation, Proliferation, Migration, Invasion
图1 NOD样受体家族蛋白3在正常子宫内膜组织中的免疫组化检测情况（SP法，×400）
图2 NOD样受体家族蛋白3在子宫内膜样腺癌组织中的免疫组化检测情况（SP法，×400）
图3 正常子宫内膜组织及不同分化程度子宫内膜样腺癌组织中NOD样受体家族蛋白3表达水平的比较
图4 正常子宫内膜组织及不同TNM分期子宫内膜样腺癌组织中NOD样受体家族蛋白3表达水平的比较
表1 子宫内膜样腺癌组织和正常子宫内膜组织中NLRP3表达水平的比较（±s）
组别 例数 NLRP3 mRNA NLRP3蛋白
子宫内膜样腺癌组织 13 1.02±0.21 1.32±0.25
正常子宫内膜组织 12 0.18±0.06 0.20±0.09
t值   3.769 4.316
P值   <0.01 <0.01
图5 正常子宫内膜组织和子宫内膜样腺癌组织中的NLRP3蛋白印迹条带图
表2 NLRP3在两种子宫内膜样腺癌细胞系中表达水平的比较（±s）
组别 NLRP3 mRNA NLRP3蛋白
ishikawa细胞 0.65±0.07 0.53±0.06
HEC-1A细胞 0.91±0.15 0.86±0.12
t值 2.373 2.559
P值 <0.05 <0.05
表3 下调NLRP3对ishikawa细胞和HEC-1A细胞迁移和侵袭的影响（个，±s）
组别 细胞迁移 细胞侵袭
ishikawa细胞 HEC-1A细胞 ishikawa细胞 HEC-1A细胞
对照siRNA 100.0±5.6 100.0±4.1 100.0±7.8 100.0±7.1
NLRP3 siRNA 45.0±4.9 56.0±5.7 64.0±5.3 67.0±6.9
t值 7.343 6.571 3.859 3.289
P值 <0.01 <0.01 <0.01 =0.01
图6 下调NOD样受体家族蛋白3对子宫样腺癌ishikawa细胞增殖的影响
图7 下调NOD样受体家族蛋白3对子宫样腺癌HEC-1A细胞增殖的影响
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