자궁경부암 마우스모델에서 포토론을 이용한 광역동치료의 효과

Abstract

Photodynamic therapy is a non-invasive treatment method that irradiates light after administration of a photosensitizer, and can be applied regardless of the type or stage of cancer. Photodynamic therapy experiments for cervical cancer were mostly conducted in vitro studies using cervical cancer cell lines, and there are very few in vivo experiments. The purpose of this study was to observe the effect of photodynamic therapy on cervical cancer using a human cervical cancer cell line (CaSki cell) mouse model (tumor bearing mouse model). Cultured CaSki cells were injected into mice (SCID/ICR mice), and mice with carcinoma in the back of the mice were used as a CaSki cancer bearing mouse model. Photodynamic therapy was performed by irradiating 80J/cm2 and 100J/cm2 of a laser beam (wavelength 660nm) 2 hours after intraperitoneal injection of Photolon (5mg/kg BW) into CaSki cancer bearing mice, respectively. After 24 hours of laser irradiation, the experimental animals are sacrificed and the tumor tissue is excised. A part of the tumor tissue was used as tissue material such as H&E staining and TUNEL assay, and the rest of the tumor tissue was used for biochemical tests such as 8-OHdG, TBARS and Caspase-3 assay. As a result of photodynamic therapy using the photosensitizer Photolon in CaSki cancer bearing mouse model, the tumor volume of the photodynamic therapy 80J/cm2 group was increased by 213% after 21 days of laser irradiation compared to before laser irradiation. The tumor volume of the photodynamic therapy 100J/cm2 group was reduced by 48% after 21 days of laser irradiation compared to before laser irradiation. Tumor tissue H&E staining in the light microscopy findings, the tumor tissue of the 80J/cm2 group of photodynamic therapy had an irregular nucleus, and a nucleus with an unclear nuclear membrane was observed. In the tumor tissue of the photodynamic therapy 100J/cm2 group, the irregular shape of the nucleus morphology was more severe than that of the 80J/cm2 group, and there were more cells with unclear nuclear boundaries. After 24 hours of photodynamic therapy, the percentage of TUNEL-stained nuclei in the tumor tissue of the 80J/cm2 group was 47.2%. In the photodynamic therapy 100J/cm2 group, the number of TUNEL-stained nuclei in the tumor tissue was 75.9%, and the ratio of TUNEL positive cells was 28% higher than that of the 80J/cm2 group. After 24 hours of photodynamic therapy, the 8-OHdG content in the tumor tissue was increased by 287% in the photodynamic treatment 80J/cm2 group compared to the control group. Tumor tissue 8-OHdG content was increased by 133% in the photodynamic treatment 100J/cm2 group compared to the photodynamic treatment 80J/cm2 group. Tumor tissue TBARS contents was increased by 192% in the photodynamic treatment 80J/cm2 group than in the control group. Tumor tissue TBARS content in the photodynamic treatment 100J/cm2 group was increased by 64% compared to the photodynamic treatment 80J/cm2 group. Tumor tissue Caspase-3 activity was increased by 141% in the photodynamic treatment group of 80J/cm2 compared to the control group. The tumor tissue Caspase-3 activity of the photodynamic treatment 100J/cm2 group was increased by 41% compared to the photodynamic treatment 80J/cm2 group. As a result of the above experiment, it was shown that when photodynamic therapy was performed using the photosensitizer photoron in the CaSki cancer bearing mouse model, the volume of the tumor was reduced and thus there was an anticancer effect. Reactive oxygen generated by photochemical reaction induces oxidative damage to DNA and cell membranes, and Cytochrome C is leaked due to decreased mitochondrial membrane function, thereby activating the intrinsic apoptosis pathway, which is thought to induce cancer cell death. Photodynamic therapy using Photolon and CaSki tumor-bearing mouse model is a preclinical experimental model, and additional research is needed to apply it as a clinical cancer treatment method.