Experiment Research Of The New Photosensitizer Photodynamic Therapy In The Treatment Of Cholangiocarcinoma

ObjectiveThis study,at the cellular level and live animals, explore the effect of a new type of chlorophyll photosensitizer, HCE6, in photodynamic therapy for cholangiocarcinoma cell lines QBC939 and photodynamic treatment of tumor-bearing nude mice, and explore the dose-response relationship.MethodsCCK8 test the QBC939 line cholangiocarcinoma cell viability under the treatement of HCE6 photodynamic therapy,at 652nm different laser light intensity (0.6J/cm2,1.8J/cm2,2.7J/cm2,5.4J/cm2)and at different concentrations photosensitizers (0μg/ml,0.3μg/ml,0.5μg/ml,1.0μg/ml,15μg/ml,2.0μg/ml), and find the optimal light intensity and photosensitizer concentration of inhibiting cholangiocarcinoma cell. And compare with the light toxicity and dark ness toxicity in the optimal laser dose.Establish a nude mouse model of QBC939 line cholangiocarcinoma cells,these nude mice were divided into control group and the PDT group, both groups were intravenously injected photosensitizers 0.5mg/kg.After 24h,photodynamic group received laser treatment, while the control group was not given laser treatment, after the laser treatment,at theld、3d、5d、7d、9d、11d、13d、15d,we observed the tumor volume of experimental group (PDT group) and control group and compared the tumor size between experimental group and control group.At the 15d,all the mice were killed, whichever tumors fixed in 10% formalin, embedded in wax dip, sliced, and HE staining.And compared the tumor pathological changes through the light microscopy.ResultsThe optimum light intensity dose is 2.7J/cm2, and the optimal photosensitizer concentration is 1.5μg/ml; in this light dose and photosensitizer concentration, increasing light intensity and photosensitizer concentration, cholangiocarcinoma cell viability decreased gradually,however over the light intensity dose and photosensitizer concentration,cell viability has no statistical differences,compared to the cell viability when the light intensity and photosensitizer concentration dose was optimal (p> 0.05).In nude mouse model experiments, the experimental group and the control group has significant differences in tumor volume size(p<0.05),no matter what time after experimental treatement.The 15th day after photodynamic therapy, the volume between experimental group and the control group has significant differences(p<0.05),the experimental group tumor volume size being 0.5-/+0.010cm3, while the control group 2.25-/+0.015cm3.ConclusionWhether cellular level or vivo experiments, having shown that received the new photosensitizer HCE6 photodynamic therapy, growth rate of QBC939 line cholangiocarcinoma cell was inhibited obviously.