The Study Of Pancreatic Cancer PANC-1 Cells Apoptosis Induced By AgNPs/LRME

Objectives:Nanosized silver particles（Ag NPs）refer to silver monoliths with a particle size of nanometer scale（1-100 nm）.In recent years,the studies of green synthesizing Ag NPs with plant extracts and their anticancer activity have become a hotspot.However,there are no reports about green synthesis of Ag NPs from Lycium ruthenicum Murr extract（LRME）and the effects on pancreatic cancer.Therefore,we first investigated the synthesis of Ag NPs/LRME,in addition,we further focused on the underlying mechanism of Ag NPs/LRME induced pancreatic cancer PANC-1 cells apoptosis.Research methods:First,Lycium ruthenicum Murr extract was mixed with Ag NO₃solution to synthesize Ag NPs/LRME under microwave irradiation.Ag NPs/LRME were characterized by UV-vis spectrophotometer（UV-vis）,electron beam lithography,transmission electron microscope,energy dispersive X-ray spectrometer（EDX）,X-ray diffractometer,dynamic light scattering（DLS）as well as FTIR microspectroscopy.Then the following experiments were performed after Ag NPs/LRME（75μg/m L）treatment of PANC-1 cells for 48 h:CCK-8 assay was used to detect the cell proliferation rate;AO/PI staining assay was used to detect the number of viable cells;Flow cytometry was used to detect mitochondrial membrane potential,reactive oxygen species,cell cycle arrest and cell apoptosis rate;UV spectrophotometry was used to detect MDA,SOD and lactic acid;Colorimetric method was used to detect the level of Caspase-3;ELISA method was used to detect the levels of Bax,Caspase-9,Cyt-c,Bcl-2and PARP;Agarose gel electrophoresis was used to detect DNA fragmentation;FTIR microspectroscopy was used to analyze the changes of intracellular biomacromolecules.Results:Ag NPs/LRME was successfully synthesized.UV spectrum showed the maximum absorption peak at 489 nm;SEM and TEM revealed that Ag NPs/LRME was spherical;EDX and XRD confirmed the presence of silver monoliths in Ag NPs/LRME;DLS showed that the average size of Ag NPs/LRME was 97 nm.Ag NPs/LRME were cytotoxic to PANC-1 cells through cell cycle arrest and Bax/Bcl-2 dysregulation,which in turn caused MMP decline,Cytochrome C release as well as Caspase activation,ultimately leading to PANC-1 cell apoptosis.Moreover,Ag NPs/LRME treatment caused increased intracellular ROS and MDA levels with decreased SOD levels,indicating that oxidative damage occurred in PANC-1 cells to induce apoptosis.The results of FTIR microspectroscopic study further confirmed that Ag NPs/LRME induced the damage to DNA,proteins and lipids in PANC-1 cells.Conclusions:Ag NPs/LRME was successfully synthesized in this study.Moreover,Ag NPs/LRME induced apoptosis through cycle arrest as well as mitochondrial pathway.Biomacromolecular changes during apoptosis were further revealed by FTIR spectroscopy at molecular level,which provided a useful supplement for further studying the mechanism of Ag NPs-induced pancreatic cancer cells apoptosis.