Institute for Biotechnology and Bioengineering, Centre of Biological Engineering, Universidade do Minho, Campus de Gualtar, 4710-057 Braga, Portugal.
J Med Microbiol. 2011 Sep;60(Pt 9):1270-1275. doi: 10.1099/jmm.0.031195-0. Epub 2011 May 12.
The aim of this study was to investigate the interaction of Candida tropicalis with three different human cell lines: TCC-SUP (epithelial cells from urinary bladder), HeLa (epithelial cells from cervical carcinoma) and Caco-2 (epithelial cells from colorectal adenocarcinoma). In particular we sought to assess the degree of cell damage and activity reduction induced by C. tropicalis adhesion and the role of secreted aspartyl proteinase (SAP) gene expression in this process. Two C. tropicalis strains were used: the reference strain ATCC 750 and a clinical isolate from urine (U69). The ability of C. tropicalis to adhere to a confluent layer of human cells was determined using an adaptation of the crystal violet staining method; cell damage and cell activity inhibition induced by the adhesion of C. tropicalis were assessed by measuring lactate dehydrogenase and tetrazolium salt (MTS) reduction, respectively. C. tropicalis SAP gene expression was determined by real-time PCR. Both C. tropicalis strains were able to adhere to the different human cells, although in a strain- and cell-line-dependent manner. Concerning the cellular response to C. tropicalis, the highest inhibition of cell activity was obtained for Caco-2, followed by TCC-SUP and HeLa cells. The highest percentage of cell damage (around 14 %) was observed for TCC-SUP cells in contact with the U69 isolate and for Caco-2 in contact with the reference strain. Real-time PCR analysis revealed a wide range of expression profiles of SAP genes for both C. tropicalis strains in contact with the different types of epithelial cells. SAPT3 was the gene expressed at the highest level for both C. tropicalis strains in contact with the three human epithelial cell lines. The results highlight that the response of human cells to C. tropicalis adhesion, as well as production of SAPs, is dependent on both the strain and the epithelial cell line.
本研究旨在探讨热带假丝酵母菌（Candida tropicalis）与三种不同的人类细胞系（TCC-SUP[膀胱上皮细胞]、HeLa[宫颈癌细胞]和 Caco-2[结直肠腺癌细胞]）的相互作用。我们特别评估了 C. tropicalis 黏附引起的细胞损伤和活性降低程度，以及分泌天冬氨酸蛋白酶（SAP）基因表达在这一过程中的作用。使用了两种 C. tropicalis 菌株：参考菌株 ATCC 750 和来自尿液的临床分离株 U69。采用结晶紫染色法的改良方法测定 C. tropicalis 黏附到汇合层人类细胞的能力；通过测量乳酸脱氢酶和四唑盐（MTS）还原，分别评估 C. tropicalis 黏附引起的细胞损伤和细胞活性抑制。通过实时 PCR 确定 C. tropicalis SAP 基因表达。两种 C. tropicalis 菌株均能黏附于不同的人类细胞，但黏附能力因菌株和细胞系而异。就 C. tropicalis 对细胞的反应而言，Caco-2 细胞的活性抑制最高，其次是 TCC-SUP 和 HeLa 细胞。与 U69 分离株接触的 TCC-SUP 细胞和与参考菌株接触的 Caco-2 细胞的细胞损伤百分比最高（约 14%）。实时 PCR 分析显示，两种 C. tropicalis 菌株与不同类型的上皮细胞接触时，SAP 基因的表达谱差异很大。对于两种 C. tropicalis 菌株与三种人类上皮细胞系接触时，SAPT3 是表达水平最高的基因。结果表明，人类细胞对 C. tropicalis 黏附的反应以及 SAP 的产生，既依赖于菌株，也依赖于上皮细胞系。
