The Experimental Study On The Effect And Apoptotic Mechanism Of Xibofen-PDT With ASA In Eca-109 Cell In Vitro

Background:Photodynamic Therapy (PDT) is a kind of photochemistry therapy. The PDT mechanism is that the resident photosensitizer in tumor tissue is activated by certain wavelength laser and takes action on oxygen in tumor tissue and produce some radical oxygen species (ROS), then attack the tumor cell through peroxidization. This may make the cellular membrane or oxidative protein damage. And the tumor cell will apoptosis after the oxidative damage get to the leak point. Also, it can damage to the tumor capillary. As contrast to conventional therapy such as operation and chemotherapy, PDT has the advantages as minimally invasive,little toxicity,high selectivity,wide applicability,well repeatability,be used as pamper treatment,maintain features and important organ function and can cure precancerous lesion. Nowadays, PDT has been used in curing some malignant tumor, and gradually to become a new method for tumor therapy.Esophageal carcinoma is a kind of common human alimentary canal tumor and it has high death rate in our country. As the development of the new type photosensitizers and semiconductor laser generator, PDT has made great progression in curing esophageal carcinoma especially in the earlier period and precancerous lesion. But in clinical curing, we find it is common that the photodynamic therapy effect is instability. Some human esophageal carcinoma patients can be clinical curative, while some is negative. Several laboratories have also shown that PDT can induce expression and/or activation of additional pro-angiogenic molecules including COX-2 and prostaglandins,TNF-a, matrix metalloproteinases (MMPs), integrins, IL-6,and IL-8 within the tumor microenvironment [11-18],which significantly decrease PDT. Nonsteroidal anti-inflammatory drugs (NSAIDs) exhibit antiangiogenic properties by blocking cyclooxygenase (COX) pathways that lead to the production of prostaglandins, which are powerful angiogenic mediators. Preclinical investigations indicate that combining PDT with targeted therapies directed at attenuating the pro-survival actions of the tumor microenvironment can enhance the therapeutic potential of PDT[11-15].ObjectivesThe objective of this part of experiment is to investigate the effect of different combinded ways of aspirin and PDT on the human esophageal carcinoma cell line Eca-109 and its mechanism in vitro.1,Study on the effect of different concentrations and light time of xibofen-PDT on proliferation and COX-2 expression of Eca-109 cell in vitro.Methods:1) There are four groups which contain only light, only Xibofen, PDT, and control respectively.2) Inhibitory effect of Eca-109 cell Proliferation was detected at different concentrations (0ug/ml,2.5 ug/ml,5ug/ml, 10ug/ml,15ug/ml,20 ug/ml) and light time (0s,30s,60s,90s) of xibofen-PDT with MTT.3) COX-2 andβ-actin expression of Eca-109 cell was detected at different concentrations (0 ug/ml,2.5 ug/ml,10 ug/ml,20 ug/ml) and light time (0s, 30s,60s,90s) of xibofen-PDT with In cell western blot.4) The results of experiment express by the way of mean±SD. Survival rate of each group apply the way of General linear Models of Univariate and one-way ANOVA. We defined statistical significance as P<0.05.Results:1) The effects of xibofen-PDT on proliferation:MTT assay showed that Eca-109 cell proliferation was significant inhibited by xibofen-PDT in time and dose dependent manner.2) The cell survival rate and the expression rate of COX-2/β-actin between different Xibofen incubating concentration and three different light time and under the same light time with different incubating concentrations are significant different and also significant interaction, all P<0.01.The cell survival rate is the lowest as xibofen concentration is 20.0ug/ml and the light time was 90s. The expression rate of COX-2/p-actin is the highest as xibofen concentration is 10.0ug/ml and the light time was 90s.Conclusions:1) The study showed xibofen-PDT can inhibited the proliferation of Eca-109 cells and induct apoptosis of them.2) The study also showed that related molecular mechanism of treating Eca-109 cells using xibofen-PDT were up-regulation of the expression of COX-2.2,Study on the effect of different concentrations of ASA on proliferation and COX-2 expression of Eca-109 cell in vitroMethods:1) There are two groups which contain ASA and control respectively.2) Inhibitory effect of Eca-109 cell Proliferation were detected at different concentrations(0 mmol/L,2.5 mmol/L,5 mmol/L,7.5mmol/L, 10mmol/L, 12.5mmol/L) of ASA with MTT.3) COX-2 andβ-actin expression of Eca-109 cell was detected at different concentrations (0 mmol/L,2.5 mmol/L,5 mmol/L,12.5 mmol/L) of ASA with In cell western blot.4) The results of experiment express by the way of mean±SD. Survival rate of each group apply the way of one-way ANOVA. We defined statistical significance as P<0.05.Results:1) The effects of ASA on proliferation:MTT assay showed that Eca-109 cell proliferation was significant inhibited by ASA in time dependent manner.2) The expression rate of COX-2/β-actin descent as the ASA incubating concentration increase, this means that as the incubating concentration increase,the expression rate of COX-2/β-actin of Eca-109 cells decrease.Conclusions:1) The study showed aspirin can inhibited the proliferation of Eca-109 cells,and induct apoptosis of them.2) The study also showed that related molecular mechanism of treating Eca-109 cells using aspirin were down-regulation of the expression of COX-2.3,Study on the effect of different concentrations and joint modals of xibofen-PDT with ASA on proliferation and COX-2 expression of Eca-109 cell in vitroMethods:1) There are four groups which contain PDT,PDT beofre ASA, ASA beofre PDT,and control respectively.This part is on the role of the ASA concentration(5 mmol/L) IC50 effect on Proliferation of Eca-109 cells.2) Inhibitory effect of Eca-109 cell Proliferation was detected at different concentrations (2.5ug/ml,5ug/ml, 10ug/ml,15ug/ml, 20ug/ml) and different joint modals (PDT,PDT beofre ASA,ASA beofre PDT) of xibofen-PDT and ASA with MTT.3) COX-2 andβ-actin expression of Eca-109 cell was detected at different concentrations (2.5 ug/ml,10 ug/ml,20 ug/ml) and different joint modals (PDT,PDT beofre AS A, AS A beofre PDT) of xibofen-PDT and ASA with In cell western blot.4) The results of experiment express by the way of mean±SD. Survival rate of each group apply the way of General linear Models of Univariate and one-way ANOVA. We defined statistical significance as P<0.05.Results:The cell survival rate and the expression rate of COX-2/β-actin between different Xibofen incubating concentrations and different joint modals,under the same joint modals with different incubating concentrations and under the same incubating concentration with different joint modals are significant different and also significant interaction, all P<0.01.The cell survival rate is the lowest as xibofen concentration is 20.0ug/ml and the PDT before ASA. The expression rate of COX-2/β-actin is the lowest as PDT before ASA.Conclusions:1) The study showed ASA can Enhance the killing effect of Xibofen-PDT on the Eca-109 cells.2) The study also showed that related molecular mechanism of ASA enhancing the killing effect of Xibofen-PDT on the Eca-109 cells were down-regulation of the expression of COX-2.3) Xibofen-PDT with ASA can reduce the recurrence rate of PDT.