原发性肝癌患者癌组织NLRP3炎性小体表达状况研究

doi:10.3969/j.issn.1672-5069.2021.04.023

摘要/Abstract

摘要： 目的研究原发性肝癌患者癌组织核苷酸结合寡聚化结构域富含脓素亮氨酸重复序列结构域3(NLRP3)炎性小体表达情况。方法 2017年1月～2018年1月我院诊治的原发性肝癌(PLC)患者34例和同期因肝胆管结石切除的肝组织标本30份,采用免疫组织化学法检测肝组织NLRP3炎性小体表达,采用RT-PCR法检测肝组织NLRP3 mRNA 水平,采用ELISA法检测血清IL-1β、IL-6和TNF-α水平,采用蛋白印记法检测肝组织NLRP3炎性小体、细胞钙依粘连蛋白、波形蛋白和半胱氨酸蛋白酶-1表达。结果肝癌组织细胞NLRP3炎性小体阳性率为76.5%,显著高于正常对照组的23.3%(P<0.05);肝癌组织NLRP3 mRNA水平为(-2.58±0.35),显著高于正常肝组织【(0.00±0.24),P<0.05】;PLC患者血清IL-1β、IL-6和TNF-α水平分别为(17.25±3.36) pg/ml、(60.32±8.14) pg/ml 和(24.68±3.58) pg/ml,显著高于对照组【(10.35±1.25) pg/ml、(33.21±4.20) pg/ml和(10.36±2.54) pg/ml, P<0.05】;正常对照组肝组织钙依粘连蛋白水平为(1.2±0.15),显著高于肝癌组【(0.41±0.04), P<0.05】,而半胱氨酸蛋白酶-1和波形蛋白水平为(0.21±0.03)和(0.42±0.06),显著低于肝癌组【(1.49±0.12)和(1.51±0.14), P<0.05】。结论原发性肝癌患者癌组织NLRP3炎性小体呈现高表达,可能会导致炎症反应并促进肝癌细胞的迁移。

关键词: 原发性肝癌, 核苷酸结合寡聚化结构域富含脓素亮氨酸重复序列结构域3, 炎性小体, 表达

Abstract: Objective The purpose of this study was to investigate the expression of nucleotide-binding oligomerization domain leucine rich repeats containing pyrin domain 3 (NLRP3) inflammasome in cancerous tissues of patients with primary liver cancer (PLC). Methods The tumorous specimens from 34 patients with PLC and 30 normal liver tissues from patients with cholelithiasis were obtained in our hospital between January 2017 and January 2018, and the expression of NLRP3 inflammasome were detected by immunohistochemistry. The NLRP3 mRNA levels were detected by RT-PCR. Serum IL-1β, IL-6 and TNF-α levels were assayed by ELISA. The NLRP3 inflammasome, E-cadherin, Vimentin and cysteine protease 1 (Caspase-1) expression were detected by Western blot. Results The positive rate of NLRP3 in cancerous tissues was 76.5%, significantly higher than 23.3%(P<0.05) in normal liver tissues; the NLRP3 mRNA level in cancerous tissues was (-2.58±0.35), much higher than [(0.00±0.24), P<0.05] in normal liver tissues; serum IL-1β, IL-6 and TNF-α levels in patients with PLC were (17.25±3.36) pg/ml, (60.32±8.14) pg/ml and (24.68±3.58) pg/ml, significantly higher than [(10.35±1.25) pg/ml, (33.21±4.20) pg/ml and (10.36±2.54) pg/ml, respectively, P<0.05] in patients with cholelithiasis; the expression of E-cadherin in normal liver was (1.2±0.15), significantly higher than [(0.41±0.04), P<0.05], while the hepatic expression of caspase-1 and vimentin were (0.21±0.03) and (0.42±0.06), significantly lower than [(1.49±0.12) and (1.51±0.14), respectively, P<0.05] in normal liver tissues. Conclusion NLRP3 inflammasome is highly expressed in patients with PLC, and the up-regulation might lead to inflammatory reactions and promote migration of cancer cells.

Key words: Hepatoma, Nucleotide-binding oligomerization domain leucine rich repeats containing pyrin domain 3, Inflammasome, Expression

陈维, 魏涛, 杨岚, 伍小敏. 原发性肝癌患者癌组织NLRP3炎性小体表达状况研究[J]. 实用肝脏病杂志, 2021, 24(4): 544-547.

Chen Wei, Wei Tao, Yang Lan, et al. Expression of NLRP3 inflammasome in cancerous tissues of patients with primary liver cancer[J]. Journal of Practical Hepatology, 2021, 24(4): 544-547.

参考文献

[1] Yamagu C, Kuro D, Sugitan I. Transglutaminase 2 is upregulated in primary hepatocellular carcinoma with early recurrence as determined by proteomic profiles. Int J Oncol, 2017, 50(5):1749-1751.
[2] Jia JA, Li H, Wang H, et al. Hepatitis B virus core antigen mutations predict post-operative prognosis of patients with primary hepatocellular carcinoma. J Gen Virol, 2017, 98(6):1399-409.
[3] CenLP, Liang JJ, Chen JH, et al. AAV-mediated transfer of Rhoa shRNA and CNTF promotes retinal ganglion cell survival and axon regeneration. Neuroscience, 2017, 343(2):472-482.
[4] Wang S, Lin Y, Yuan X, et al. REV-ERBα integrates colon clock with experimental colitis through regulation of NF-B/NLRP3 axis. Nat Commun, 2018, 9(1):4246-4250.
[5] Alvarez K, Vasquez G. Damage-associated molecular patterns and their role as initiators of inflammatory and auto-immune signals in systemic lupus erythematosus.Int Rev Immunol, 2017, 36(5):259.
[6] 赵静, 成军. 肝纤维化发生信号通路的研究进展. 中华肝脏病杂志, 2019, 27(6):403-406.
[7] Vogel S, Murthy P, Cui X,et al. TLR4-dependent upregulation of the platelet NLRP3 inflammasome promotes platelet aggregation in a murine model of hindlimb ischemia. Biochem Biophys Res Commun. 2019, 508(2):614-619.
[8] 王学国, 杨彦, 黎东明, 等. 微小RNA-1973对肝细胞癌细胞增殖的影响及其分子机制. 中华实验外科杂志, 2019, 36(10):1755-1757.
[9] 杨秉辉, 任正刚. 原发性肝癌诊断标准. 中华肝脏病杂志, 2000, 8(3):135-135.
[10] SanninoG, Marchetto A, Kirchner T, et al. Epithelial-to-mesenchymal and mesenchymal-to-epithelial transition in mesenchymal tumors: a paradox in sarcomas? Cancer Res,2017, 77(17):5468-5472.
[11] WreeA, Eguchi A, Mcgeough MD, et al. NLRP3 inflammasome activation results in hepatocyte pyroptosis, liver inflammation, and fibrosis in mice. Hepatology, 2014, 59(8):898-910.
[12] Bronsert P, Enderle K, Bader M, et al. Cancer cell invasion and EMT marker expression: a three-dimensional study of the human cancer-host interface. J Pathol, 2015, 234(3):410-422.
[13] Wang L, Zhao Z, Feng W, et al. Long non-coding RNA TUG1 promotes colorectal cancer metastasis viaEMT pathway. Oncotarget, 2016, 7(32):51713-517139.
[14] Wang JH, Lee EJ, Ji M, et al. HDAC inhibitors, trichostatin A and valproic acid, increaseE-cadherin and vimentin expression but inhibit migration and invasion of cholangiocarcinoma cells. Oncol Rep, 2018, 40(1):346-354.
[15] Esa SA, Rawy AM, Elbehissy MM, et al. Study the level of sputum matrix metalloproteinase-9 and tissue inhibitor metaloprotienase-1 in patients with interstitial lung diseases. Egyp J Chest Dis Tuber,2016, 65(1):303-309.
[16] Xu Y,Li H,Chen W,et al. Mycoplasma hyorhinis activates the NLRP3 inflammasome and promotes migration and invasion of gastric cancer cells. PLoS One, 2013, 8(11):e77955.
[17] Ada N, Tam WW, Zhang MW, et al. IL-1β, il-6,TNF- α and CRP in elderly patients with depression or alzheimer's disease: systematic review and meta-analysis. Sci Rep, 2018, 8(1):12050-12052.
[18] ShenC, Lu A, Xie WJ, et al. Molecular mechanism for NLRP6 inflammasome assembly and activation. Proc Natl Acad Sci U S A, 2019, 116(5): 2052-2057.
[19] ZhangNP, Liu XJ, Xie L, et al. Impaired mitophagy triggers NLRP3 inflammasome activation during the progression from nonalcoholic fatty liver to nonalcoholic steatohepatitis. Lab Invest, 2019, 99(7): 749-763.
[20] Li M, Chen Y, Shi J, et al. NLRP6 deficiency aggravates liver injury after allogeneic hematopoietic stem cell transplantation. Int Immunopharmacol, 2019, 74(11):105740-105746.