Li LinWei, Xia Jing, Cui RuTing, Kong Bin
Department of Breast Disease Center, The Affiliated Hospital of Qingdao University, No. 59 of Haier Road, Shandong 266000 Qingdao, China.
Department of Breast Surgery, Qingdao Central Hospital, Qingdao, 266000 Shandong China.
Cytotechnology. 2021 Feb;73(1):23-34. doi: 10.1007/s10616-020-00439-y. Epub 2020 Nov 11.
This study aimed to understand the expression of solute carrier family 12 member 8 (SLC12A8) in breast carcinoma and its biological functions, as well as its effect on the Toll-like receptor /NOD-like receptor (TLR/NLR) signaling pathway. The expression of SLC12A8 was analyzed using the public RNA sequencing dataset from TCGA database and the two datasets from Oncomine database. The former dataset was also used to evaluate the prognostic value of SLC12A8 in breast carcinoma. Real-time qPCR and western blot were applied to measure relative expression of SLC12A8. Functionally, the effect of SLC12A8 on the cells proliferation and motion was studied using cell counting kit 8 and Transwell assays respectively. Mechanistic studies were conducted using Gene Set Enrichment Analysis (GSEA) and confirmed by western blot. As a result, SLC12A8 was upregulated in breast carcinoma, and high levels of SLC12A8 led to a poorer prognosis and can be regarded as an independent prognosticator for patients with breast carcinoma. Functional experiments demonstrated that SLC12A8-knockdown suppressed while SLC12A8-overexpression elevated the viability, invasiveness and motility of breast carcinoma cells. Furthermore, GSEA indicated that high SLC12A8 was positively correlated with TLR/NLR signaling pathway. Silencing SLC12A8 significantly reduced the protein expression of TLR/NLR-related markers, whereas overexpression of SLC12A8 caused an elevation on the protein expression of these markers. All these data suggested that SLC12A8 plays a promoting effect on the cells viability, invasiveness and motility in breast carcinoma by activating TLR/NLR signaling pathway.
本研究旨在了解溶质载体家族12成员8(SLC12A8)在乳腺癌中的表达及其生物学功能,以及其对Toll样受体/NOD样受体(TLR/NLR)信号通路的影响。使用来自TCGA数据库的公共RNA测序数据集和来自Oncomine数据库的两个数据集分析SLC12A8的表达。前一个数据集也用于评估SLC12A8在乳腺癌中的预后价值。应用实时定量PCR和蛋白质印迹法检测SLC12A8的相对表达。在功能上,分别使用细胞计数试剂盒8和Transwell实验研究SLC12A8对细胞增殖和运动的影响。使用基因集富集分析(GSEA)进行机制研究,并通过蛋白质印迹法进行验证。结果显示,SLC12A8在乳腺癌中上调,高水平的SLC12A8导致较差的预后,可被视为乳腺癌患者的独立预后指标。功能实验表明,敲低SLC12A8可抑制而过表达SLC12A8可提高乳腺癌细胞的活力、侵袭性和运动性。此外,GSEA表明高SLC12A8与TLR/NLR信号通路呈正相关。沉默SLC12A8显著降低了TLR/NLR相关标志物的蛋白表达,而过表达SLC12A8则导致这些标志物的蛋白表达升高。所有这些数据表明,SLC12A8通过激活TLR/NLR信号通路对乳腺癌细胞的活力、侵袭性和运动性起促进作用。