FKN Facilitates HK-2 Cell EMT and Tubulointerstitial Lesions via the Wnt/ β-Catenin Pathway in a Murine Model of Lupus Nephritis
In this study, we therefore examined whether FKN could stimulate the process of EMT, NF-kB, TGFβ, CCL22, F4/80, inflammation, and tubulointerstitial fibrosis in a murine model of LN. We also determined whether FKN was involved in the EMT process of Wnt/β-catenin-expressing HK-2 cells. Mechanistically, we ascertained, for the first time, whether FKN up-regulated EMT-related gene signatures (e.g., vimentin, α-SMA), and hence, renal tubulointerstitial fibrogenesis, and the role of the Wnt/β-catenin signaling pathway in this process. Materials and Methods Cell Culture, Stable Infection, and Grouping HK-2 cells were obtained from the Cell Center of Fudan University (Shanghai, China), and cultured in DMEM/F12 medium (Gibco) with 10% fetal bovine serum (Gibco) at 37°C in a humidified 5% CO2 atmosphere. HK-2 cells were infected with lentiviral vector particle-CX3CL1, lentiviral vector particle-CX3CL1-RNAi, and hU6-MCS-Ubiquitin-EGFP-IRES-negative control according to manufacturer protocol (Shanghai Genechem Co., Ltd.) to effect FKN overexpression or knockdown (KD). After 12 h of infection, cells were cultured in fresh complete medium for 48–72 h, FKN protein expression was examined using western blotting. The cells were divided into nine groups as follows: (1) Control group; (2) FKN-KD group; (3) XAV939 group; (4) FKN-KD + XAV939 group; (5) Ex-FKN group; (6) Ex-FKN + XAV939 group; (7) Ang II group; (8) FKN-KD + Ang II group; and (9) Ex-FKN + Ang II g...
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