The Evaluation Of Matrine Lectin Modified Nanoparticles To Human Colon Cancer Cells HT29

Objective: The issue selectin preliminary study prepared by modified nanoparticles matrine(WGA-MT-NPS) as the research object, using specific lectin affinity interaction of cancer cells, colon cancer HT29 cells in the role of evaluation, WGA-MT-NPS understand the mechanism of cancer cells, colon lay the foundation for an oral bioadhesive drug delivery system targeting studies.Methods:(1) Preparation and Characterization of WGA-MT-NPS and agglutination active investigation Preparated WGA-MT-NPS, evaluated with particle size, particle size distribution and the potential, and examine the activity of after lectin-modified on matrine nanoparticles.(2) Effect of WGA-MT-NPS HT29 colon cancer cell proliferation Using MTT, OD value of inhibition to study MT-NPS WGA before and after modification HT29 colon cancer cells proliferation.(3) Effect of WGA-MT-NPS apoptosis of HT29 colon cancer cells Using Annexin V-FITC / PI double staining by flow cytometry study MT-NPS influence of the group and the WGA-MT-NPS apoptosis in HT29 cells, and using Flow Jo software for analysis.(4) Effect of WGA-MT-NPS HT29 colon cancer cells cell cycle Using PI staining and using flow cytometry study MT-NPS influence of the group and the WGA-MT-NPS on HT29 cell cycle and use ModFit software for analysis.(5) Different affinity research of WGA-MT-NPS to HT29 colon cancer cells and normal human colonic epithelial cells NCM460 Respectively react with fWGA-MT-NPS in HT29 cells and NCM460 cells using fluorescent dye staining affinity of the drug and in the case of two cell confocal microscope.Results: According to the optimal preparation process, the three batch of MT-NPS particle size were 102.6nm, 104.6nm and 101.6nm, the potential were-17.5mV,-12.2mV and-12.5 mV, the three batch of WGA-MT-NPS particle particle size were 234.7nm, 235.1nm and 231.9nm, the potential were-9.51 mV,-7.85 mV and-7.54 mV; Through investigation we know, the clotting activity did not change after the WGA modifited on MT-NPS. WGA-MT-NPS has a stronger inhibitory effect on the proliferation of HT29 cells, and has an obvious dose effect relationship. The same drug concentration of WGA-MT-NPS relative to MT-NPS, the apoptosis of colorectal cancer cell HT29 was higher. Drugs stop the cell cycle at G0/G1 phase,and then synthesis of hindered cell DNA, and in the G0/G1 phase WGA-MT-NPS cell volume more than MT-NPS. Observed under a confocal microscope, the green fluorescence intensity of colon cancer cell line HT29 in the field of obvious strong in the normal human colonic epithelial cell line NCM460.Conclusion: WGA still keeps its activity of specific sugar residues binding after modified nanoparticles; WGA-MT-NPS has stronger inhibitory and promoting apoptosis effect on the proliferation of HT29 cells than MT-NPS, as well as the cell cycle arrest action. HT29 cells was significantly stronger than normal colon epithelial cells NCM460 on WGA-MT-NPS uptaking. MT-NPS modified with WGA, showed evident targeting and selective cytotoxicity on colon cancer cells...