Study On The Killing Effect Of 5-ALA Photodynamic Therapy On Ovarian Cancer Transplanted Tumor Cells In Nude Mice

Objective:To explore the killing effect of 5-ALA photodynamic therapy(PDT)on ovarian cancer transplanted tumor cells in nude mice.Methods:The subcutaneous transplantation model of ovarian cancer in nude mice was established and divided into model group,5-ALA group,laser irradiation group and5-ALA-PDT group,with 10 mice in each group.The 5-ALA group and the 5-ALA-PDT group were given intratumoral injection of 40 mg/kg 5-ALA sterile saline solution,respectively.The model group and the laser irradiation group were given intratumoral injection of the same amount of saline solution.After 4 h of light protection,the tumor was locally irradiated with 630 nm laser therapy instrument in laser irradiation group and5-ALA-PDT group for 20 minutes each time.Once every 2 days,a total of 5times.After the treatment,the tumor volume was measured,and the mice in the four groups were killed by dislocation method for dissection,and the tumor was removed for the next experiment.(1)The size of the transplanted tumor was measured and the tumor growth curve was drawn;(2)Tumor inhibition rate was calculated and histogram was drawn for comparison;(3)Tissue sections stained with HE;(4)TUNEL staining was used to detect tumor cell apoptosis.(5)cleaved Caspase-3,Bcl-2 gene,and Bax gene m RNA were measured by q RT-PCR(real-time fluorescence quantitative PCR).(6)Protein expressions of cleaved Caspase-3,Bcl-2,and Bax were detected by Western blot.Results:Compared with the model group and the 5-ALA group,the tumor volume was lower in both the laser irradiation group and the 5-ALA-PDT group after 14 days of intervention,and the tumor volume in the 5-ALA-PDT group was the lowest among the four groups(P<0.001).The tumor inhibition rate of laser irradiation group and5-ALA-PDT group after 7 d and 14 d was higher than that of model group and 5-ALA group(P<0.001),and that of 5-ALA-PDT group was higher than that of laser irradiation group(P<0.05).Cleaved Caspase-3 and Bax m RNA expression and protein expression were higher in the laser irradiation group and the 5-ALA PDT group than in the model group and the 5-ALA group(P<0.001),and the 5-ALA-PDT group was higher than the laser irradiation group(P<0.001).The expression of Bcl-2 m RNA and protein in the laser irradiation group and the 5-ALA-PDT group were lower than those in the model group and the 5-ALA group(P<0.001),and the expression of Bcl-2 protein in the5-ALA-PDT group was lower than that in the laser irradiation group(P<0.001).Conclusion:5-ALA-PDT treatment of ovarian cancer can enhance apoptosis induction of tumor cells by up-regulating cleaved Caspase-3 and Bax expression and inhibiting Bcl-2expression,thus enhancing its killing effect.