A Study Of Vitamin D₃ Enchancing 5-ALA Mediated Photodynamic Therapy On Tumor

Photodynamic therapy(PDT)is a kind of minimally invasive therapy to treat tumor.Nowadays,PDT is applied into skin tumor and skin precancerous lesions,head and neck cancer,encephalic tumor,respiratory system tumor,digestive system tumor,urinary system tumor,gynecological oncology.PDT has many advantages as nice specificity,minimally invasive,less traumatic,protecting normal tissue,and nice repetitions.However it also has some disadvantages as the bad light penetrability,having vestigial when treat the large tumor and relapse;the patience who treat with the PDT need avoid the sunlight a week.5-ALA is the pro drug to PpIX.5-ALA is the second generation photosensitizer.5-ALA compares with other photosensitizers is more sable,better photodynamic effect and less side effects.Vitamin D3 is a kind of vitamin D group,and it is the most activity of vitamin D group.The vitamin D3 distributes wildly in human body,it can adjust the metabolism of calcium and phosphorus,and maintaining bones and teeth development.The studies manifest that vitamin D3 can inhibit tumor growth,promote tumor apoptosis,inducing tumor differentiation,regulation the immune system,and enhancing sensibility to chemotherapy and radiotherapy.Recently study reveal that vitamin D3 can enhance the PDT curative effect in vitro that haematoporphyrin as the photosensitizer.To investigate whether the vitamin D3 enhance PDT curative effect or not.We studied the SK-BR-3 cell lines and MCF-7 cell lines in vitro and in vivo.The two cell lines are come from human breast cancer.We examined whether PDT combined vitamin D3 will enhance PDT curative effect or not.In vitro studies,we used MTT assay measure every group's OD value.We obtained the cell inhabitation rate by computed each group OD value.The results showed vitamin D3 combined PDT cell inhabitation is markedly higher than PDT group(SK-BR-3 cell lines cell inhabitation rate 75.82%�0.74 vs.47.04%�1.86;MCF-7 cell lines cell inhabitation rate 88.61�5.14 vs.76.76�3.18).We obtained cell apoptosis rate by FCM(flow cytometry).The results showed that vitamin D3 combined PDT group cell apoptosis rate is higher than PDT group(SK-BR-3 cell apoptosis rate 38.13%�2.15vs.18.20%�2.07,MCF-7cellapoptosis rate 63.14%�3.55vs.30.53%�6.30);Then,the SK-BR-3 cell lines and the MCF-7 cell lines were be incubation in 5-ALA and 5-ALA + D3 24 hours,and used multifunction mircoplate reader to measure each group cell fluorescence intensity.The results showed 5-ALA+D3 group fluorescence intensity was higher than 5-ALA group(SK-BR-3 cell lines fluorescence intensity 193.80�15.18 vs.114.80�21.31,MCF-7 cell fluorescence intensity 299.10�20.76vs.118.50�16.70).This study imply that vitamin D3 can enhance PDT,and increase cell inhabitation rate and cell apoptosis rate by increase intracellular 5-ALA concentration.In vivo study,subcutaneous into tumor in nude mouse left breast by injected MCF-7 cell line.PDT+D3 group mice were be intrapersonal injection vitamin D3 lug/kg 3 days before PDT treatment.Then,each group get corresponding treatment and carry out observations.After treatment 24 hours random executed a mouse from each group.The lesion tissues were obtained by the mice.These tissues were be fixation,embedding and section.These tissue were stained by H.E and immunohistochemical.The PDT+D3 group lesion has more eschar and less tumor residual tumor than PDT group lesion.HE staining showed most of PDT + D3 group tissue' cell nucleus are pyknosis and disappear,and most of PDT group tissues' cell nucleus pyknosis,several cell nucleus fragmentation.Immunohistochemical staining showed PDT + D3 group BAX protein expression is more than PDT group,PDT + D3 group tissue VEGF-A protein expression is less than PDT group.We observed the mice lifetime,PDT + D3 group mice median lifetime were 24�4.38 day,PDT group mice median lifetime were 18�2.19 day.These results suggest PDT combined vitamin D3 improve PDT curative effect in vivo,increase mice tumor apoptosis,inhibition vessel growth in tumor,and prolong mice lifetime.