原发性肝癌患者癌组织NLRP3炎性小体表达状况研究

Expression of NLRP3 inflammasome in cancerous tissues of patients with primary liver cancer

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。

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-αlevels 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-αlevels 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.