| Objective: The antitumor activities and mechanism of compounds perylene bisimidecyclodextrin-carbohydrate conjugates(PBI-2TMCD-Man and PBI-2TMCD-3Man)were investigated through MTT method,flow cytometry and fluorescence imaging.The topic will provide the basis for the research and development of new photosensitizers.Method:1.Antitumor activities of PBI-2TMCD-Man and PBI-2TMCD-3Man in vitroThe inhibitory effects of PBI-2TMCD-Man and PBI-2TMCD-3Man and positive control cisplatin(non-photosensitizer)and TPPS(photosensitizer)on cell proliferation of four cell lines were detected by MTT;The phototoxicity and dark toxicity of PBI-2TMCD-Man and PBI-2TMCD-3Man were evaluated in both light and non-irradiated conditions.2.Confocal imaging of PBI-2TMCD-Man and PBI-2TMCD-3Man on cellsMCF-7 and Hep G2 cell lines were treated with PBI-2TMCD-Man and PBI-2TMCD-3Man separately for 24 h,then were carried out without irradiation and illumination for 12 h.The changes of the cells entering the cell and the morphological changes were observed by laser confocal microscopy.3.Apoptosis induced by PBI-2TMCD-Man and PBI-2TMCD-3ManThe sensitive cell lines(MCF-7,Hep G2)of compounds were selected and treated for 24 h after irradiation for 12 h and 24 h.All cells were stained with Annexin V-FITC / PI and measured by flow cytometry.4.Effects of PBI-2TMCD-Man and PBI-2TMCD-3Man on cell cycleThe sensitive cell lines(MCF-7,Hep G2)of compounds were treated for 24 h,then were carried out without irradiation,illumination for 12 h and 24 h.All cells were stained with PI single staining and measured by flow cytometry.Result:1.The results of MTT assay showed that PBI-2TMCD-Man and PBI-2TMCD-3Manshowed higher phototoxicity than that of the two positive controls,and the dark toxicity was low.Compared with no irradiation,the IC50 of PBI-2TMCD-Man for four tumour cells decreased from 182.54 ?M,315.78 ?M,208.40 ?M,69.33 ?M to 3.96 ?M,16.79 ?M,6.62 ?M,5.14 ?M with illumination for 0.5 h.The IC50 of PBI-2TMCD-3Man for four tumour cells decreased from 174.08 ?M,220.63 ?M,166.48 ?M,41.89 ?M to 5.78 ?M,2.41 ?M,5.35 ?M,1.65 ?M with illumination for 0.5 h.The IC50 of TPPS for four tumour cells decreased from 65.55 ?M,44.00 ?M,155.38 ?M,30.57 ?M to 40.17 ?M,27.81 ?M,23.20 ?M,22.82 ?M with illumination for 0.5 h.The IC50 of cisplatin for four tumour cells were 9.54 ?M,9.66 ?M,12.98 ?M,6.60 ?M.2.After tumor cells were treated by both compounds,apoptosis of morphological changes such as cell size becomes smaller,cells out of the bubble were observed by laser confocal microscopy.Both PBI-2TMCD-Man and PBI-2TMCD-3Man enter the cytoplasm but not enter the nucleus.3.After tumor cells were treated by both compounds,the effects of PBI-2TMCD-Man and PBI-2TMCD-3Man on tumor cells were detected by flow cytometry(Annexin V-FITC /PI).Compared with no irradiation,PBI-2TMCD-Man's apoptotic rate increased from 3.91%to 84.56%.PBI-2TMCD-3Man's apoptotic rate was increased from 11.94% to 80.01%,and their apoptotic rates were significantly higher than cisplatin and TPPS.4.PBI-2TMCD-Man blocked the cell cycle in G1 / G0 phase and PBI-2TMCD-3Man blocked the cell cycle in S period.Conclusion:1.The phototoxicity of PBI-2TMCD-Man and PBI-2TMCD-3Man on four tumour cells were stronger than cisplatin and TPPS.The dark toxicity of them on four tumour cells were lower than cisplatin and TPPS.Their biological activities were stronger than cisplatin and TPPS.2.PBI-2TMCD-Man was the most sensitive to MCF-7 and PBI-2TMCD-3Man had the best inhibitive activity for Hep G2.3.Both PBI-2TMCD-Man and PBI-2TMCD-3Man cause cell volume reduction,nuclearcondensation and other morphological changes.Both PBI-2TMCD-Man and PBI-2TMCD-3Man could enter the cytoplasm and could not enter the nucleus.4.The photodynamic effect of both compounds were enhanced with prolonged illumination time.5.The onset time of PBI-2TMCD-3Man was shorter than that of PBI-2TMCD-Man.6.PBI-2TMCD-Man could block MCF-7 cells in the G1 / G0 phase and PBI-2TMCD-3Man could block Hep G2 cells in the S phase.Both of them induced apoptosis. |
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