To investigate the interaction mechanism of casein (CA) and whey protein (WP) with anthocyanins from purple potato flour (PPA), multiple spectroscopic techniques and molecular docking simulation were used. After extraction and purification, two non-acylated anthocyanins and three acylated anthocyanins were identified in purple potato flour using UPLC-Q/TOF-MS. The fluorescence quenching experiment found that PPA could effectively quench the intrinsic fluorescence of CA and WP by static quenching. CA had a stronger binding affinity toward PPA than WP did, with respective K-a values of 4.23 x 10(5) M-1 and 1.66 x 10 5 M-1 at 297 K. Structural analysis indicated that PPA bound CA and WP through hydrogen bonds and van der Waals forces, and the number of bound anthocyanin molecules (n) was approximately equal to 1. The AG values of PPA binding with CA and WP were -30.96 kJ mol(-1) and -29.16 kJ mol(-1), respectively, which suggested that the binding reaction was spontaneous. Moreover, the conformational structures of CA and WP were altered by PPA binding with a decrease in alpha-helix and beta-turn contents and an increase in beta-sheet and irregular coil contents. Molecular docking analyses showed that CA and WP had different binding sites with anthocyanins. This study is helpful to better understand the interaction mechanism of CA and WP with anthocyanins and provides guidance for their applications in the food industry.

Background: Circular RNAs (circRNAs) play crucial roles in the development and progression of human cancers, including non-small cell lung cancer (NSCLC). However, most of these circRNAs, such as hsa_circ_0014235, are not fully identified in functions and mechanisms. Methods: The isolated exosomes from serum specimens were identified using transmission electron microscopy (TEM). The expression of hsa_circ_0014235, miR-520a-5p and cyclin-dependent kinase 4 (CDK4) was detected by real-time quantitative polymerase chain reaction (qPCR). For functional assays, cell proliferation, colony formation ability, migration, invasion, cell apoptosis and cell cycle progression were determined using cell counting kit-8 (CCK-8) assay, colony formation assay, wound healing assay, transwell assay and flow cytometry assay, respectively. The expression of CDK4 and other indicated marker proteins was detected by western blot. The predicted target relationship between miR-520a-5p and hsa_circ_0014235 or cyclin-dependent kinase 4 (CDK4) was verified by dual-luciferase reporter assay or RNA immunoprecipitation (RIP) assay. Results: The expression of hsa_circ_0014235 was notably elevated in NSCLC serum-derived exosomes, tumor tissues and cells. NSCLC serum-derived exosomes promoted NSCLC cell resistance to cisplatin (DDP), cell proliferation, migration and invasion in vitro, as well as tumor growth and DDP resistance in vivo. Hsa_circ_0014235 overexpression enhanced DDP resistance and facilitated cell malignant behaviors. MiR-520a-5p was a target of hsa_circ_0014235, and rescue experiments showed that miR-520a-5p restoration reversed the effects of hsa_circ_0014235 overexpression. Moreover, CDK4 was a target of miR-520a-5p, and rescue experiments showed that CDK4 knockdown reversed the aggressive effects of miR-520a-5p inhibition on NSCLC progression. Conclusions: Exosome-transmitted hsa_circ_0014235 promoted NSCLC malignant development by mediating the miR-520a-5p/CDK4 regulatory axis.