The Effects Of Different Molecular Weight HGAG With HPD-PDT On The Proliferation Inhibition And Apoptosis Of Human Lung Adenocarcinoma A549 Cells

Objective Explore the effects of different molecular weight sea cucumber glycosaminoglycan(HGAG)combined with photodynamic therapy(HPD-PDT)on proliferation inhibition and apoptosis of human lung adenocarcinoma A549 cells,so as to provide theoretical support for exploring new clinical treatments for lung cancer.Methods Select human lung adenocarcinoma A549 Cells are the research object,and they are grouped by in vitro culture to an appropriate degree: Control group(cells +culture medium,no other treatment),3W molecular weight sea cucumber glycosaminoglycan group(3W molecular weight sea cucumber glycosaminoglycan concentration was 100μg/ml),5W molecular weight sea cucumber glycosaminoglycan group(5W molecular weight sea cucumber glycosaminoglycan concentration was100μg/ml),photodynamic group(hematoporphyrin derivative concentration was 8μg/ml and irradiated by 630 nm laser),3W molecular weight sea cucumber glycosaminoglycan combined with photodynamic group(100μg/ml of 3W molecular mass sea cucumber glycosaminoglycan/ml combined with 8μg/ml hematoporphyrin derivative and irradiated by 630 nm laser),5W molecular weight sea cucumber glycosaminoglycan combined with photodynamic group(100μg/ml of 5W molecular mass sea cucumber glycosaminoglycan combined with 8μg/ml hematoporphyrin derivative and irradiated by630 nm laser),After the treatment,the cells in each group were continued to be cultured,detected and data collected:(1)Observe the morphological changes of cells in each group under a microscope;(2)Determine the cell survival rate and calculate the interaction coefficient(CDI)by CCK-8 experiment;(3)Stain each group by Hoechst33258 stain,Fluorescence microscopy was used to observe;(4)Eachl group was stained by Annexin V-FITC/PI double staining method,and the apoptosis rate was detected by flow cytometry.Results(1)Microscopically,after HGAG and HPD-PDT treatment,the cell count in the culture medium decreased,the cells were sparse and less dense,and a few floating cells were visible on the liquid surface,among which the two combined groups were the most obvious;(2)The results of the CCK-8 assay showed that both HGAG and HPD-PDT could inhibit the growth of human lung adenocarcinoma A549 cells,and HGAG could promote the growth inhibition of A549 cells by HPD-PDT and improve the sensitivity of HPD-PDT.Compared with the single drug group,the inhibition rate of cell proliferation in each combination group increased(all P < 0.05);(3)Hochest 33258 staining experiment showed that nuclear fragmentation could be observed under microscope in HGAG group,HPD-PDT group and combination group The two combined groups were the most obvious and the least dense;(4)Annexin V-FITC/PI double staining combined with flow cytometry showed that the experimental groups were significantly different from the control group.There were statistically significant differences in the total apoptotic rate among the experimental groups(all P < 0.05).Conclusion Different molecular weights of HGAG have obvious inhibitory effects on the growth and proliferation of human lung adenocarcinoma A549 cells,and the molecular weight of 3W is better,which can enhance the sensitivity of A549 cells to HPD-PDT.