NLRP3炎症小体在食管鳞状细胞癌中的表达及意义

Expression and Significance of NLRP3 Inflammasome in Esophageal Squamous Cell Carcinoma

目的分析NOD样受体热蛋白结构域相关蛋白3(NOD-like receptor pyrin domain containing 3,NLRP3)炎症小体在食管鳞状细胞癌(esophageal squamous cell carcinoma,ESCA)的表达及意义。方法临床研究:选取2021年5月—2023年5月山西省汾阳医院收治的168例ESCA患者为研究对象。患者均经过手术治疗,使用免疫组织化学及蛋白印迹法测定癌旁组织、ESCA组织的NLRP3、生长分化因子11(growth differentiation factor 11,GDF11)蛋白表达。动物及细胞研究:将BALB/c裸鼠按处理方式的不同分为对照组、GDF11过表达慢病毒载体组(GDF11组),记录并比较所有组别裸鼠7、14、21、28 d的体质量及30 d的瘤体积。筛选ESCA细胞(人食管鳞状细胞癌细胞系ES2、人食管鳞状细胞癌细胞系TE-1、人食管鳞状细胞癌细胞系RJEC-2、人食管鳞状细胞癌细胞系KYSE-170)NLRP3较高表达及GDF11较低表达的细胞系为研究对象,采用转染技术将ESCA细胞分为过表达对照组、GDF11过表达组、敲减对照组和GDF11敲减组,比较各组的细胞增殖能力。结果临床研究:168例ESCA组织中NLRP3阳性123例(73.21%),阴性45例(26.79%);GDF11阳性35例(20.83%),阴性133例(79.17%)。癌旁组织中NLRP3阳性26例(15.48%),阴性142例(84.52%);GDF11阳性126例(75.00%),阴性42例(25.00%)。ESCA组织NLRP3蛋白表达水平高于癌旁组织,GDF11蛋白表达水平低于癌旁组织,差异有统计学意义(P<0.001)。NLRP3、GDF11在ESCA组织、癌旁组织表达差异有统计学意义(P<0.05)。ESCA组织内NLRP3、GDF11呈负相关(P<0.001);GDF11阴性表达的ESCA组织的NLRP3为(1.12±0.18),高于GDF11阳性表达ESCA组织(0.80±0.14),差异有统计学意义(P<0.001)。动物及细胞研究:GDF11组裸鼠7、14、21、28 d的体质量高于对照组,差异有统计学意义(P<0.05)。GDF11组裸鼠瘤体积为(73.26±21.25)mm^(3),低于对照组[(137.28±45.28)mm^(3)],差异有统计学意义(P=0.003)。GDF11过表达组2、3、4、5 d的ESCA细胞增殖能力低于过表达对照组,GDF11敲减组2、3、4、5 d的ESCA细胞增殖能力高于敲减对照组,差异有统计学意义(P<0.05)。结论NLRP3炎症小体在食管癌呈高表达,通过GDF11调节NLRP3炎症小体或成为治疗ESCA的潜在治疗策略。

Objective To analyze the expression and significance of NOD-like receptor pyrin domain containing 3(NLRP3)inflammasome in esophageal squamous cell carcinoma(ESCA).Methods Clinical study:a total of 168 ESCA patients admitted to Fenyang Hospital,Shanxi Province from May 2021 to May 2023 were selected as the study objects.After surgical treatment,NLRP3 and growth differentiation factor 11(GDF11)protein expression in paracancer tissue and ESCA tissue were measured by immunohistochemistry and Western Blot.Animal and cellular research:BALB/c nude mice were divided into control group and GDF11 overexpressing lentiviral vector group(GDF11 group)according to different treatment methods.The body weight at 7,14,21 and 28 days and tumor volume at 30 days of all groups were recorded and compared.The cells with high NLRP3 expression and low GDF11 expression in ESCA cells(human esophageal squamous cell carcinoma cell line ES2,human esophageal squamous cell carcinoma cell line TE-1,human esophageal squamous cell carcinoma cell line RJEC-2,human esophageal squamous cell carcinoma cell line KYSE-170)were selected as the study objects.ESCA cells were divided into overexpression control group,GDF11 overexpression group,knockdown control group and GDF11 knockdown group by transfection technique,and the cell proliferation ability of each group was compared.Results Clinical study:Among 168 ESCA tissues,123 cases(73.21%)were positive for NLRP3 and 45 cases(26.79%)were negative.GDF11 was positive in 35 cases(20.83%)and negative in 133 cases(79.17%).NLRP3 was positive in 26 cases(15.48%)and negative in 142 cases(84.52%).GDF11 was positive in 126 cases(75.00%)and negative in 42 cases(25.00%).The expression level of NLRP3 protein in ESCA tissues was higher than that in paracancer tissues,and the expression level of GDF11 protein was lower than that in paracancer tissues,with statistical significance(P<0.001).There were significant differences in the expressions of NLRP3 and GDF11 in ESCA tissues and para-cancerous tissues,with statistical s ignificance(P<0.05).In ESCA,NLRP3 and GDF11 were negatively correlated(P<0.001).The NLRP3 of ESCA tissue with negative expression of GDF11 was(1.12±0.18),which was higher than that of ESCA tissue with positive expression of GDF11(0.80±0.14),with statistical significance(P<0.001).Animal and cellular research:the body weight of nude mice in GDF11 group at 7,14,21 and 28 days was higher than that in control group,and the difference was statistically significant(P<0.05).The tumor volume of nude mice in GDF11 group was(73.26±21.25)mm^(3),which was lower than that in control group(137.28±45.28)mm^(3),with statistical significance(P=0.003).The proliferation capacity of ESCA cells at 2,3,4 and 5 days in the GDF11 overexpression group was lower than that in the overexpression control group,and the proliferation capacity of ESCA cells at 2,3,4 and 5 days in the GDF11 knockdown group was higher than that in the knockdown control group,with statistical significance(P<0.05).Conclusion NLRP3 inflammasome is highly expressed in esophageal cancer,and the regulation of NLRP3 inflammasome by GDF11 may be a potential therapeutic strategy for ESCA.