Liang Shan, Xiang Tingting, Liu Shiyu, Xiang Wei
College of Modern Agriculture and Bioengineering, Yangtze Normal University, Chongqing 408000, P.R. China.
Institute of Sericulture and Systems Biology, Southwest University, Chongqing 400700, P.R. China.
Exp Ther Med. 2022 Jul 28;24(3):601. doi: 10.3892/etm.2022.11538. eCollection 2022 Sep.
Gastric cancer (GC) is one of the commonest malignant tumors of the digestive system, characterized by high morbidity and mortality rates. It has been reported that NOD like receptor (NLR) family, CARD domain containing 5 (NLRC5) serves an important role in the occurrence and development of GC. Therefore, the current study aimed to investigate the role of NLRC5 in GC. The mRNA and protein expression levels of NLRC5 in GC cell lines were determined by reverse transcription-quantitative PCR and western blot analysis, respectively. Additionally, following NLRC5 knockdown, cell proliferation, invasion and migration were evaluated using Cell Counting Kit 8, colony formation, wound healing and Transwell assays, and western blot analysis. The NLRC and Yin Yang 1 (YY1) expression in the AGS cells with 5-FU resistance were detected by western blotting. The sensitivity of GC cells to 5-fluorouracil (5-FU) was detected by flow cytometry and western blot analysis. Additionally, the binding capacity of YY1 on NLRC5 promoter was predicted using JASPAR database and it was further verified by chromatin immunoprecipitation and luciferase reporter assays. Finally, to elucidate the mechanism underlying the effect of NLRC5 on GC, YY1 was overexpressed and NLRC5 was silenced in GC cell lines. The results showed that NLRC5 was abnormally upregulated in GC cells. In addition, NLRC5 knockdown significantly attenuated the proliferation, invasion and migration abilities of GC cells, while it enhanced the sensitivity of GC cells to 5-FU. The above effects were regulated by the YY1 transcription factor. Overall, the results of the present study indicated that NLRC5 silencing could reduce the malignant growth and enhance the sensitivity of GC cells to 5-FU chemotherapy via inhibiting the carcinogenic effect of YY1.
胃癌(GC)是消化系统最常见的恶性肿瘤之一,其发病率和死亡率都很高。据报道,含CARD结构域的NOD样受体(NLR)家族成员5(NLRC5)在胃癌的发生发展中起重要作用。因此,本研究旨在探讨NLRC5在胃癌中的作用。分别采用逆转录定量PCR和蛋白质免疫印迹法检测NLRC5在胃癌细胞系中的mRNA和蛋白表达水平。此外,在敲低NLRC5后,使用细胞计数试剂盒8、集落形成、伤口愈合和Transwell实验以及蛋白质免疫印迹分析评估细胞增殖、侵袭和迁移能力。通过蛋白质免疫印迹法检测AGS细胞中5-氟尿嘧啶(5-FU)耐药时NLRC和阴阳1(YY1)的表达。通过流式细胞术和蛋白质免疫印迹分析检测胃癌细胞对5-氟尿嘧啶(5-FU)的敏感性。此外,使用JASPAR数据库预测YY1对NLRC5启动子的结合能力,并通过染色质免疫沉淀和荧光素酶报告基因实验进一步验证。最后,为阐明NLRC5对胃癌作用的机制,在胃癌细胞系中过表达YY1并沉默NLRC5。结果显示,NLRC5在胃癌细胞中异常上调。此外,敲低NLRC5可显著减弱胃癌细胞的增殖、侵袭和迁移能力,同时增强胃癌细胞对5-FU的敏感性。上述作用受YY1转录因子调控。总体而言,本研究结果表明,沉默NLRC5可通过抑制YY1的致癌作用降低胃癌细胞的恶性生长并增强其对5-FU化疗的敏感性。