Xue Lan, Yang Lu, Fu Xize, Feng Wenli, Yang Jing, Ma Yan, Xi Zhiqin
Department of Dermatology, The Second Hospital of Shanxi Medical University, Taiyuan, Shanxi, 030001, China.
Department of Dermatology, The Second Hospital of Shanxi Medical University, Taiyuan, Shanxi, 030001, China.
Biochem Biophys Res Commun. 2025 Jun 14;777:152147. doi: 10.1016/j.bbrc.2025.152147.
The link between SAP2 and Candida albicans' capacity to adhere to and invade host cells is notable, although the precise molecular mechanisms remain unclear. The purpose of this study was to examine how elevated SAP2 expression influences the invasion and adhesion capabilities of C. albicans in A431 cells using in vitro experiments and transcriptomic sequencing.
The effect of high SAP2 expression on the invasion of A431 cells by C. albicans was evaluated using lactate dehydrogenase (LDH) release assay. Morphological changes in hyphae were observed under a microscope during co-culture with A431 cells. The ability of C. albicans to adhere to A431 cells was assessed using an adhesion assay. Transcriptomic sequencing and qRT-PCR were used to examine changes in gene expression and associated signaling pathways in C. albicans after co-culture with A431 cells.
After 1.5-2 h of co-culture, the number of A431 cell colonies adhered by the SC5314 strain was significantly higher than adhered by the SAP2-overexpressing SC5314 strain (pRB895-SAP2-SC5314). After 8 h, the SC5314 strain induced the formation of dense, elongated hyphae, whereas the pRB895-SAP2-SC5314 strain formed clustered, snowflake-like yeast forms. After 24 h, the LDH release assay revealed a significantly higher death rate of A431 cells in the SC5314 group. Differential expression analysis identified 50 upregulated and 10 downregulated genes in the pRB895-SC5314 strain relative to the SC5314 strain. A total of 1142 genes were upregulated and 1219 genes were downregulated in the pRB895-SAP2-SC5314 strain relative to the SC5314 strain. In addition, 1159 genes were upregulated and 1228 genes were downregulated in the pRB895-SAP2-SC5314 strain relative to the pRB895-SC5314 strain. KEGG enrichment analysis indicated the MAPK signaling pathway associated with these genes, and qRT-PCR showed significant downregulation of genes such as TEC1.
Co-culture with A431 cells significantly inhibited hyphal growth and attenuated the invasion and adhesion abilities of the pRB895-SAP2-SC5314 strain. Several genes related to SAP2 were upregulated or downregulated, suggesting that the role of SAP2 in the pathogenic mechanisms of C. albicans is influenced by complex regulatory factors, with the MAPK signaling pathway serving as a key factor.
尽管确切的分子机制尚不清楚，但SAP2与白色念珠菌黏附并侵袭宿主细胞的能力之间的联系值得关注。本研究的目的是通过体外实验和转录组测序，研究SAP2表达升高如何影响白色念珠菌在A431细胞中的侵袭和黏附能力。
使用乳酸脱氢酶（LDH）释放试验评估高SAP2表达对白色念珠菌侵袭A431细胞的影响。在与A431细胞共培养期间，在显微镜下观察菌丝的形态变化。使用黏附试验评估白色念珠菌黏附A431细胞的能力。转录组测序和qRT-PCR用于检测与A431细胞共培养后白色念珠菌基因表达和相关信号通路的变化。
共培养1.5 - 2小时后，SC5314菌株黏附的A431细胞集落数显著高于过表达SAP2的SC5314菌株（pRB895 - SAP2 - SC5314）。8小时后，SC5314菌株诱导形成致密、细长的菌丝，而pRB895 - SAP2 - SC5314菌株形成簇状、雪花状的酵母形态。24小时后，LDH释放试验显示SC5314组中A431细胞的死亡率显著更高。差异表达分析确定，相对于SC5314菌株，pRB895 - SC5314菌株中有50个基因上调，10个基因下调。相对于SC5314菌株，pRB895 - SAP2 - SC5314菌株中共有1142个基因上调，1219个基因下调。此外，相对于pRB895 - SC5314菌株，pRB895 - SAP2 - SC5314菌株中有1159个基因上调，1228个基因下调。KEGG富集分析表明这些基因与MAPK信号通路相关，qRT-PCR显示TEC1等基因显著下调。
与A431细胞共培养显著抑制了pRB895 - SAP2 - SC5314菌株的菌丝生长，并减弱了其侵袭和黏附能力。几个与SAP2相关的基因上调或下调，表明SAP2在白色念珠菌致病机制中的作用受复杂调控因子影响，其中MAPK信号通路是关键因素。
