| The objective of the study was to compare the killing effects of photodynamic therapy on Hela cells and the Hela/Taxol cells resistant to Taxol,in order to investigate the efficacy of photodynamic therapy in treating cervical cancer and cervical cancer resistant to chemotherapy.1.Establishment of Hela/Taxol Cell Line for Human Cervical Cancer Resistance to Taxol and Its ResistanceMethods:?Hela cells of human cervical cancer,as parentage cells,were selected by progressive method by adding taxol to the culture medium.?The cell morphology and growth trends of Hela and Hela/Taxol cells were observed and growth curves were drawn.? Detection of half inhibition concentration(IC50)of Hela and Hela/Taxol cells by MTT method.?Expression of polydrug-resistant gene P-gp,GST-? and apoptosis-related gene Bcl-2 and Survivin in Hela and Hela/Taxol cells were detected by RT-PCR.?The expression of Bcl-2,P-gp,GST-? and Survivin proteins in Hela and Hela/Taxol cells were detected by immunofluorescence and protein printing.Results:Compared to Hela cells,Hela/Taxol cells have the following characteristics.? Slow growth.? The IC50 value of Hela/Taxol increased significantly under different chemotherapy drugs.TheTh resistance to Taxol is strongest and to 5-FU is lowest.The results showed that Hela/Taxol cells had stronger drug resistance.?The gene and protein of Resistance genes P-gp and GST-? expression increased.?The gene and protein of Bcl-2 and survivin expression increased.2.Study on the killing effect of photodynamic therapy on Hela cells and Hela/Taxol cellsMethods:?Experimental Group:The Hela and Hela/Taxol cells were treated with blank control group,simple photosensitizer group,simple light group,photosensitizer with light group.?MTT Method for Detection of Inhibitory Effect of Photodynamic Therapy on Hela and Hela/Taxol Cell Lines.The optimal combination parameters of ALA concentration and light energy were selected.? The optimal combination parameters of ALA concentration and light energy were selected.?Detection of cell cycle and apoptosis of Hela and Hela and Taxol cells before and after photodynamic action by flow cytometry.?Expression of P-gp,GST-?,Bcl-2 and Jianceguang protein in two cells before and after immunofluorescence therapy.?Naked rat tumefaction experiment.Results:?Hela/Taxol cells are more sensitive to photodynamic therapy than Hela cells.?ALA concentration lmmol/L?Light energy 5J/cm2 is ideal conditions for killing Hela and Hela/Taxol cells in vitro for photodynamic therapy??Photodynamic therapy can significantly induce apoptosis of Hela and Hela/Taxol cells.And inhibit the growth of the expression of protein P-gp,GST-?,Bcl-2 and Survivin.?The apoptosis rate in both Hela-PDT and Hela/Taxol-PDT groups increased significantly.?The experimental results of nude mouse tumorigenesis show that:Hela-PDT and Hela/Taxol-PDT cell-forming tumors grow more slowly than those formed by Hela and Hela/Taxol cells,resulting in small,lightweight tumors.So photodynamic therapy can inhibit the growth of subcutaneous transplanted tumors in nude mice,and it can inhibit the formation of tumor body in Hela/Taxol cells.Conclusion:Photodynamic therapy significantly inhibits proliferation of Hela and Hela/Taxol cells in vitro,promoting apoptosis.Hela/Taxol is more sensitive to photodynamic therapy than Hela cells,and its mechanism may be related to inhibition of protein expression in P-gp,GST-?,Bcl-2 and survivin.Innovation:Hela/Taxol cell lines resistant to paclitaxel have been developed and their resistance has been demonstrated,providing an important research vehicle for future research on resistant tumors.Although Hela/Taxol cells are resistant,when photodynamic therapy is applied to both cells,the inhibition of Hela/Taxol is higher and the killing effect more evident... |
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