KLF5通过调控NTSR1促进脂肪酸氧化抑制胃癌细胞失巢凋亡的机制研究

Abstract

Neurotensin receptor 1 (NTSR1) is linked with poor prognosis in many tumors, including gastric cancer (GC). However, the exact mechanism by which NTSR1 affects GC anoikis in GC has not yet been clarified. Therefore, this project aimed to figure out the specific mechanism by which NTSR1 participated in GC anoikis. We herein assessed the expression of KLF5 and NTSR1 in GC tissues and the signaling pathways in which NTSR1 participated through the online bioinformatics website. Quantitative reverse transcription polymerase chain reaction (qRT-PCR) was employed to examine the expression of KLF5 and NTSR1. The CCK-8 kit was applied to detect the viability of GC cells under different treatments. Dual luciferase and chromatin immunoprecipitation (ChIP) experiments verified the binding relationship between KLF5 and NTSR1. The rate of fatty acid oxidation (FAO) was analyzed using a cell metabolism meter. The expression of FAO-related proteins was detected using the western blot (WB). The anoikis of cells in each group was detected using flow cytometry, anoikis apoptosis kit, and WB. We observed that knocking down NTSR1 repressed the vitality of GC cells, and elevated the anoikis rate and the expression of cleaved PARP and cleaved caspase 3 in GC cells. Moreover, high expression of NTSR1 up-regulated the expression of CTP1 protein in FAO and increased FAO levels, thereby suppressing the occurrence of anoikis in GC cells. The addition of the inhibitor of FAO (Etomoxir) reversed the above trends. In addition, KLF5 was highly expressed in GC, and KLF5 is an upstream transcription factor of NTSR1. Finally, KLF5 knockdown was verified to restore the repression of the FAO pathway and facilitate anoikis in GC cells by overexpressing NTSR1. In summary, our results suggested that KLF5 affects anoikis in GC cells by targeting NTSR1 to modulate the FAO pathway.