Establishment Of Hela/Taxol Cell Lines Resistant To Cervical Cancer And Study Of Photodynamic Therapy To Kill Hela And Hela/Taxol Cells

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:①Hela cells of human cervical cancer,as parentage cells,were selected by progressive method by adding taxol to the culture medium.②The cell morphology and growth trends of Hela and Hela/Taxol cells were observed and growth curves were drawn.③ Detection of half inhibition concentration(IC50)of Hela and Hela/Taxol cells by MTT method.④Expression of polydrug-resistant gene P-gp,GST-π and apoptosis-related gene Bcl-2 and Survivin in Hela and Hela/Taxol cells were detected by RT-PCR.⑤The expression of Bcl-2,P-gp,GST-π 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.① Slow growth.② 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.③The gene and protein of Resistance genes P-gp and GST-π expression increased.④The 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:①Experimental Group:The Hela and Hela/Taxol cells were treated with blank control group,simple photosensitizer group,simple light group,photosensitizer with light group.②MTT 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.③ The optimal combination parameters of ALA concentration and light energy were selected.④Detection of cell cycle and apoptosis of Hela and Hela and Taxol cells before and after photodynamic action by flow cytometry.⑤Expression of P-gp,GST-π,Bcl-2 and Jianceguang protein in two cells before and after immunofluorescence therapy.⑥Naked rat tumefaction experiment.Results:①Hela/Taxol cells are more sensitive to photodynamic therapy than Hela cells.②ALA concentration lmmol/L、Light energy 5J/cm2 is ideal conditions for killing Hela and Hela/Taxol cells in vitro for photodynamic therapy。③Photodynamic therapy can significantly induce apoptosis of Hela and Hela/Taxol cells.And inhibit the growth of the expression of protein P-gp,GST-π,Bcl-2 and Survivin.④The apoptosis rate in both Hela-PDT and Hela/Taxol-PDT groups increased significantly.⑤The 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-π,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...