Experimental Study Of A New Photosensitizer Based On Polyamine Transport System For The Treatment Of Pancreatic Cancer

Objective To prove that pancreatic cancer cell line PANC-1 can transport the fluorescent molecule 4-dimethylamino-1,8-naphthalimide-spermine derivative(N2),synthesized by our group,into cells through polyamine transport system(PTS).Subsequently,a photosensitizer spermine derivative(N3),which can be transported into cells through PTS,was synthesized.DFMO can promote N3 uptake by PANC-1 cells.N3 has low dark toxicity and good phototoxicity to PANC-1 cells at nanomole concentration.This study will provide new methods and ideas for the treatment of pancreatic cancer and other polyaminedependent cancers.Methods(1)The fluorescence intensity was detected by flow cytometry and automatic microplate reader.The transport mode of fluorescent molecule N2 into the cell was analyzed.(2)The photosensitizer spermine derivative(N3)was synthesized.The purity of N3 was measured by HPLC and the structural analysis of N3 was measured by NMR and Mass spectrometry.(3)The ultraviolet absorption spectra,emission and excitation spectra of N3 were detected by UV-vis spectrophotometry and fluorescence spectrophotometry.(4)Inverted fluorescence microscope and automatic microplate reader were used to analyze the N3 uptake of PANC-1 cells and the promoting effect of DFMO on the uptake process.(5)The dark toxicity of N3 and the phototoxicity of N2 and N3 were evaluated by CCK-8.Results(1)Pancreatic cancer cell line PANC-1 transport N2 into cells through polyamine transport system.(2)The photosensitizer spermine derivative(N3)was successfully synthesized and its purity and structure were analyzed.(3)The maximum ultraviolet absorption wavelength of N3 was 540 nm,the maximum excitation wavelength was around 540 nm,and the maximum emission wavelength was about 560 nm.The wavelength range of excitation and emission for follow-up experiments is determined.(4)The uptake of N3 by PANC-1 cells was in a time and concentration-dependent manner,and DFMO could promote N3 uptake by PANC-1 cells.(5)N3 had almost no toxicity to PANC-1 cells at the experimental concentration.When PANC-1 cells were treated with nanomole N2 and N3 for PDT,the cell survival rate was proportional to the concentration of N3,while the fluorescent molecule N2 had no obvious phototoxicity to the cells.Conclusions It is proved that the spermine derivative synthesized by our group can be transported into the cell by PTS which is highly expressed on the cell membrane of pancreatic cancer cell line PANC-1,and DFMO can promote the uptake of spermine derivatives by cells.The photosensitizer spermine derivative(N3)has good phototoxicity on pancreatic cancer by PDT after acting on PANC-1 at nanomole level.The successful implementation of this project will provide new methods and ideas for the treatment of pancreatic cancer and other polyamine-dependent cancers.