Clinical Effect And Experimental Study On Esophageal Carcinoma By Photodynamic Therapy Combined With Chemotherapy

The morbidity of esophageal carcinoma is so prevalent that it is the ninth of the world,and the mainly accidence area includes Asia.While one half of the esophageal carcinoma accurs in our country,and 97.6%pathology of the carcinoma is leprose cell.Different stages takes on various treatments.Surgery therapy is the treatment of choice for the early esophageal carcinoma,however the recrudescent rate is high. Moreover almost one half of the patients are dignosed advanced esophageal carcinoma initially,losing the opportunity of surgery,further more some patients can't endure operation.Chemotherapy is also one of the important theatment for esophageal carcinoma,however multidrug resistance(MDR)and severe adverse reaction limit its effect.The rapid growth of the nowadays photodynamic therapy(PDT) is a new treatment for the esophageal carcinoma patients,which can lightening dysphagia,improving the patients quality life,and prolonging ones' life. PDT also has the opportunity of little indury to physical body,finer selective cure, little adverse reaction,and normal orgen function after PDT.For the low KPS patients, advanced patients or carcinoma in situ,PDT are a good select.Many studies indicate that PDT for esophageal carcinoma takes on fine curative effect.PDT will cure small early esophageal carcinoma,but can hardly heal advanced esophageal carcinoma. PDT combined with chemotherapy would possibly gain a better effect.At present,the photosensitizers for the treatment of tumors mainly include Hematoporphyrin Photosensitizer,Acetyl-amino Propionate,Chlorin Porphyrin Thiophene,Benzo Porphyrin Derivatives,Texaphyrin,etal.Hematoporphyrin Photosensitizer was the first developed and listed HPD drugs for tumor.Among them, Photofrin(porfimer sodium),Canada AXCAN company produced,has been FDA approved for the treatment of various tumors and Barrett's esophagus lesions(BE) with severe dysplasia(HGD) since 1996,and is already registered in more than 10 countries.The drug is a mixture of Oligomeric porphyrin,and achieve a positive effect in the diagnosis and treatment of tumors.The purpose of the study was to investigate the superior to the means of HPD-PDT combined with chemotherapy for esophageal carcinoma,and the mechanism of cell and molecule.In the first part,PDT with chemotherapy on the advanced esophageal carcinoma was retrospective studied,to compare the effect of PDT vs PDT combined with chemotherapy.In the second part,the mechanism of PDT combined with chemotherapy to esophageal carcinoma Eca-109 cell line experimentally was explored,which may be advisory in combined therapy of clinical esophageal carcinoma.Chapter One Retrospective Study of Photodynamic Therapy Combined with Chemotherapy on the Advanced Esophageal CarcinomaObjective To compare the curative effect of photodynamic therapy(PDT) vs PDT combined with chemotherapy on the advanced esophageal cancer,discuss the superiority of PDT combined with chemotherapy for esophageal cancer. Method Retrospective analysis sixty patients of esophageal cancer underwent PDT or PDT combined with chemotherapy from the years of 2002 to 2005(Ⅲstages toⅣstages),including 27 cases received PDT alone,and 33 cases received PDT combined with chemotherapy.PDT used Photofrin 2mg/kg,and after 48 hours the location of lesion were irradiated by 630nm-laser light under the endoscopic guidance. Chemotherapy projects were 5-FU clus DDP for complex treatment patients, administered four cycles after PDT.Result All the 60 patients were followed up for 2 months to 56months. Symptomatic palliation rates of the complex treatment groups and the PDT alone group are separately 93.9%and 85.2%,and effective rates by the endoscopic are separately 90.9%and85.2%,there are no statistically significant difference;and survival rates of two years are separately 42%and 26%,and medium survival time is longer(stagesⅢ,33m vs 17m;stagesⅣ8m vs3m),the sequent effect of Bgroup is better than A by Cox Regression.Conclusion PDT combined with chemotherapy for the advanced esophageal cancer is superior to PDT alone,with longer survival time of two years.Chapter Two Experimental Study on Esophageal carcinoma Eca-109 Cell Line by Photodynamic Therapy Combined with ChemotherapyObjective To investigate the effects of Photodynamic Therapy(PDT) combining chemotherapy drugs(5-Fu and DDP) on esophageal carcinoma Eca-109 cells line. Methods Esophageal neoplasm cells line Eca-109 were divided six groups of A,B, C,D,E and F.A group:control group,B group:chemotherapy,C group:high HPD dose PDT+chemotherapy,D group:low HPD dose PDT+chemotherapy,E group: high HPD dose PDT,F group:low HPD dose PDT.Esophageal carcinoma cells including chemotherapy group cells were incubated in vitro with chemotherapy drugs(5-Fu50ug/ml and DDP 1.5ug/ml) for 12 hours,and then they were incubated in vitro with HPD in high(1.5ug/mL) or low(0.375 ug/mL) concentrations for 4 hours. PDT was then performed on these treated cells with 630nm laser,optical energy density of 30J/cm~2.Before and after the exposure,the cells were observed and taken photos under the inverted microscope and fluorescence microscope.The cells continued to cultivate after 24h.Proliferation inhibition rate of the cells were detected by MTT method,apoptosis rate by flow cytometry,and apoptosis-related protein Bcl-2 expression by Western blot.Results1.The observation of morphological changeUnder Inverted microscope,all of the experimental group cells with photosensitizer lh after PDT:the experimental group cells had different levels of morphological changes,cell rounded,volume shrinking,but the structural integrity of cell membranes.Adherent cells shrinkage,rounded,shedding.After 4h these changes became more obvious.There were more abnormal cells in high HPD dose PDT combined chemotherapy group.And in the control group,cells were adhering in good condition,polygonal,larger,cytoplasm fullness,bordering clearly.2.Fluorescent light intensity changes in cells before and after PDTUnder fluorescent microscope,all the experimental group cells with the photosensitizer were fluorescent,the higher the concentration of photosensitizer,the cells showed the stronger fluorescence;After PDT the fluorescent light significantly weakened.While the control group cells showed no fluorescence.3.Proliferation inhibition rate of the cells by MTT methodWith analysis of variance,the inhibition rates among all the groups showed difference(F=140.674,P=0.000).With Games-Howell,the results showed that there were no statistic difference between the low HPD dose PDT combined with chemotherapy group and the high HPD dose PDT group,and there were significant difference,among the other groups.High HPD dose PDT cells inhibited the higher rate,low HPD dose PDT cells inhibition rate lower.4.Apoptosis rate by flow cytometryAnalysis of variance under completely random design showed that A～F groups apoptosis rates were statistic different(F=481.588,P=0.000).With LSD conducted by multiple groups,the results showed that in addition to chemotherapy group with the high HPD dose PDT group difference not statistic,the other differances were significant.The High HPD dose PDT group had the highest rate of apoptosis,and control group had the lowest apoptosis rate.5.Protein Bcl-2 expression by Western blotProtein electrophoresis showed that:high HPD dose PDT combined with chemotherapy group color was shallow,deep color of the control group.Colors of the experimental groups to the control group were varying degrees shallowing.Conclusion1.Under the special light source and saturated lighten ergydensity,the photosensitizer concentration is the main factor of PDT effect on human esophageal carcinoma cells in vitro.The cells' inhibitions of two different concentrations of HPD were different. Under the same light energy density and incubation time,the higher the concentration of HPD,the stronger effect of its destruction.2.Photodynamic combined with chemotherapy could increase the lethality of esophageal carcinoma cells,increase the rate of apoptosis,and decrease protein Bcl-2 expression.PDT could synergy with chemotherapy to promote tumor cells apoptosis by channels of inhibiting expression of protein Bcl-2.3.HPD can well be excited by 410-450nm wave length of the blu-ray to show fluorescent effect.The higher concentration of HPD,and the more photosensitizer intook by esophageal cancer cells,the cells showing the stronger fluorescence effect. Under the same concentration of the HPD,the strength of the fluorescence is proportional to the activity,HPD can be used for fluorescence diagnosis.