敲低NOD2通过调控(p)NF-κB/STAT3改善肝细胞癌细胞炎症反应  

Knockdown of NOD2 improves inflammatory response in hepatocellular carcinoma cells by regulating (p)NF-κB/STAT3

作　　者：赵亮[1] 艾尔哈提·胡赛音[1] 布祖克拉·阿布都艾尼 王锦秋 亚力坤·赛来[1] ZHAO Liang;AIERHATI Husaiyin;BUZUKELA Abduaini;WANG Jinqiu;YALIKUN Sailai(Department of General Medicine,The First Affiliated Hospital of Xinjiang Medical University,Urumqi 830054,China;The Second Department of Intensive Care,The First Affiliated Hospital of Xinjiang Medical University,Urumqi 830054,China)

机构地区：[1]新疆医科大学第一附属医院全科医学科(普通外科),新疆乌鲁木齐830054 [2]新疆医科大学第一附属医院重症二科,新疆乌鲁木齐830054

出　　处：《西部医学》2023年第7期964-969,共6页Medical Journal of West China

基　　金：新疆维吾尔自治区自然科学基金面上项目(2021D01C309)。

摘　　要：目的 探讨核苷酸结合寡聚化结构域蛋白2(NOD2)对肝细胞癌(HCC)细胞炎症反应的影响和机制。方法 培养HCC细胞。Western blot检测NOD2在HCC细胞和正常肝细胞中的表达水平。利用小干扰RNA(siRNA)建立NOD2的敲低RNA(siNOD2组)用来抑制NOD2的表达,阴性对照为siNC组,使用这二者处理HCC细胞。CCK-8法检测细胞增殖率,流式细胞术检测细胞凋亡率。Western blot检测NF-κB P65、STAT3、NOD2的表达,并检测磷酸化的(p-)NF-κB P65以及磷酸化的(p-)STAT3的表达水平。ELISA法测定培养HCC细胞培养物上清液中TNF-α、IL-12、IL-6、IFN-γ质量浓度的变化。在siNOD2处理HCC细胞的基础上,用NF-κB/STAT3的激活剂重组人Lipocalin-2(rhLipocalin-2)蛋白进行处理(siNOD2+rhLipocalin-2组),检测细胞增殖率、凋亡率和炎症因子水平。结果 与正常肝细胞相比,HCC细胞中NOD2的表达水平显著上调(P<0.05)。与siNC组相比,siNOD2组的NOD2、p-NF-κB P65及p-STAT3的表达水平均显著下调、细胞增殖率降低,细胞凋亡率增高,且细胞培养物上清液中TNF-α、IL-12、IL-6、IFN-γ水平均下调(均P<0.05)。而与siNOD2组相比,siNOD2+rhLipocalin-2组的p-NF-κB P65及p-STAT3的表达水平均显著上调,细胞增殖率增高,细胞凋亡率降低,且细胞培养物上清液中TNF-α、IL-12、IL-6、IFN-γ水平均上调(均P<0.05)。结论 敲低NOD2通过调控NF-κB/STAT3通路抑制HCC细胞的炎症反应,该结果为HCC治疗提供了一个新的潜在靶点。Objective To investigate the effect and mechanism of inhibiting the activation of nucleotide binding oligomerization domain protein 2(NOD2)on inflammatory response of hepatocellular carcinoma cells.Methods Hepatocellular carcinoma cells were cultured.Western blot was used to detect NOD2 expression in hepatocellular carcinoma cells and normal hepatocytes.Small interfering RNA(siRNA)was used to establish NOD2 knockdown agents(siNOD2 group)to inhibit NOD2 expression,while the negative control group was siNC group,and hepatocellular carcinoma cells were treated.Cell proliferation rate was measured by CCK-8 method and apoptosis rate was measured by flow cytometry.The expressions of NF-κB P65,STAT3 and NOD2 were detected by Western blot,and the expression levels of phosphorylated(P-)NF-κB P65 and phosphorylated(P-)STAT3 were detected.The concentrations of TNF-α,IL-12,IL-6 and IFN-γin supernatant of hepatocellular carcinoma cell culture were determined by ELISA.On the basis of siNOD2 treatment,hepatocellular carcinoma cells were treated with NF-κB/STAT3 activator recombinant human lipocalin-2 protein(siNOD2+RHlipocalin-2 group).The proliferation rate,apoptosis rate and inflammatory factor levels of hepatocellular carcinoma cells were measured.Results Compared with normal hepatocytes,NOD2 expression level in hepatocellular carcinoma cells was significantly up-regulated(P<0.05).Compared with siNC group,NOD2,P-NF-κB P65 and P-STAT3 expression levels in siNOD2 group were significantly down-regulated(all P<0.05),cell proliferation rate was decreased(P<0.05),and cell apoptosis rate was increased(P<0.05).The levels of TNF-α,IL-12,IL-6 and IFN-γin supernatant of cell culture were down-regulated(all P<0.05).Compared with siNOD2 group,the expression levels of P-NF-κB P65 and P-STAT3 in siNOD2+RHlipocalin-2 group were significantly up-regulated(all P<0.05),the cell proliferation rate was increased(P<0.05),and the cell apoptosis rate was decreased(P<0.05).The levels of TNF-α,IL-12,IL-6 and IFN-γin supernatant of cell cul

关 键 词：肝细胞癌 核因子-κB/信号转导与转录激活因子3 核苷酸结合寡聚化结构域蛋白2 炎症反应 

分 类 号：R735.7[医药卫生—肿瘤] R392.1[医药卫生—临床医学]