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目的:研究ALA、HPD及ALA联合HPD光动力学治疗C6胶质瘤细胞瘤基因P16及P53变化。方法:以不同剂量ALA(20μg/ml、40μg/ml、60μg/ml、80μg/ml、160μg/ml)、HPD(1μg/ml、2μg/ml、3μg/ml、4μg/ml、5μg/ml)及不同剂量组合ALA联合HPD光动力学治疗C6胶质瘤细胞,以流式细胞仪测试基因P16及基因P53的变化。结果:本实验中未作PDT治疗的对照组,肿瘤细胞生长良好,可见P16值小于6.59%。P53值小于4%。而PDT治疗组随着光敏剂剂量的加大P16及P53基因值逐渐上升,是未作PDT治疗组的3-4倍,且两光敏剂混合组,P16及P53基因值大于单纯ALA或HPD-PDT治疗组。HPD1μg/ml联合ALA60μg/ml,HPD2μg/ml联合ALA40μg/ml就能达到HPD5μg/ml PDT治疗组P16基因及P53基因水平。结论:推测PDT疗法激活了P16基因及p53基因功能,促使肿瘤细胞凋亡及坏死。联合疗法HPD1μg/ml联合ALA60μg/ml,HPD2μg/ml联合ALA40μg/ml光动力学治疗能达到HPD常规剂量5μg/ml PDT的效果,这样可明显提高ALA-PDT疗效及减少HPD-PDT的皮肤光毒反应。
OBJECTIVE: To study the changes of P16 and P53 gene in C6 glioma by ALA, HPD and ALA combined with HPD photodynamic therapy. Methods: HPD (1μg / ml, 2μg / ml, 3μg / ml, 4μg / ml and 5μg / ml) were treated with different doses of ALA (20μg / ml, 40μg / ml, 60μg / ml, 80μg / ml, 160μg / The C6 glioma cells were treated with different doses of combined ALA and HPD photodynamic therapy. The changes of P16 gene and P53 gene were detected by flow cytometry. Results: The control group without PDT in this experiment, tumor cells grew well, showing P16 value of less than 6.59%. P53 value is less than 4%. PDT treatment group with the dose of photosensitizer increased P16 and P53 gene value is not PDT treatment group 3-4 times, and the two photosensitizer mixed group, P16 and P53 gene values greater than the ALA alone or HPD- PDT treatment group. HPD1μg / ml with ALA60μg / ml, HPD2μg / ml with ALA40μg / ml can reach HPD5μg / ml PDT treatment group P16 gene and P53 gene levels. Conclusion: PDT therapy is presumed to activate the function of P16 gene and p53 gene and promote the apoptosis and necrosis of tumor cells. Combination therapy HPD1μg / ml with ALA60μg / ml, HPD2μg / ml combined with ALA40μg / ml photodynamic therapy can achieve the effect of conventional dose of HPD 5μg / ml PDT, which can significantly improve the efficacy of ALA-PDT and reduce skin phototoxicity of HPD-PDT reaction.