Simvastatin-loaded PCL/PEG nanofibrous scaffold: A prospective approach for suppression 5-fluorouracil resistance in MKN-45 gastric cancer cells

Abstract

BackgroundBecause the development of chemoresistance is a key issue in the treatment of gastric cancer (GC), there has been significant interest in combination chemotherapy. In this study, we examined whether simvastatin-loaded Polycaprolactone– Polyethylene Glycol nanofibers (SIM-PCL/PEG NFs) inhibit 5-fluorouracil (5-FU) resistance in MKN-45 gastric cancer cells. Methods5-FU-resistant MKN-45 cells (MKN-45/R) were established by increasing concentrations of SIM. SIM-PCL/PEG NFs were synthesized and characterized through SEM and FTIR testing. SIM-PCL/PEG NFs cytotoxicity on MKN-45 and MKN-45/R cells was determined using the MTT assay. mRNA expression of P53, Bax, Bcl-2, ULK1, Beclin1, LC3, MMP2, and MMP9 genes in both cell groups was measured by RT-PCR. Flow cytometry, DAPI staining, and colony formation assay were used to assess the anti-proliferative activity of SIM-PCL/PEG NFs on cells. Autophagy-related proteins and cell migration were evaluated using western blotting and wound healing assay, respectively. The STRING database was used for bioinformatics analyses. ResultsMTT assay showed that SIM-PCL/PEG NFs had a more cytotoxic effect on MKN-45/R cells. Flow cytometry analysis indicated that SIM-PCL/PEG NFs could cause more cell apoptosis in MKN-45/R cells and also, the mRNA expression levels showed significantly elevated p53 and Bax levels and reduced Bcl-2 in MKN-45/R cells in this group. Analysis of ULK1, Beclin1, and LC3 at the transcription and translation levels indicated SIM-loaded NFs promote autophagy in MKN-45/R cells. Furthermore, compared with the control group colony formation and migration of MKN-45/R cells in the SIM-loaded NFs group were significantly inhibited (all P < 0.05). ConclusionWe have introduced simvastatin-loaded nanofibres as a promising therapeutic compound for the inhibition of 5-FU resistance in gastric cancer cells.