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目的:探讨HPPH光动力治疗对鼠G422脑胶质瘤突变型p53蛋白和p16蛋白表达的影响并和HpD做比较。方法:建立鼠G422脑胶质瘤皮下移植瘤模型,设立空白对照组、单注射HPPH组(0.45mg/kg)、单665nm激光照射组、HPPH-PDT组(0.15mg/kg组、0.3mg/kg组、0.45mg/kg组)、HpD-PDT组(5mg/kg)。单注射HPPH组、HPPH-PDT组和HpD-PDT组自尾静脉推注入光敏剂,24h后以波长665nm的半导体激光照射HPPH-PDT组和单激光组肿瘤,功率密度200mW/cm~2,每光斑照射17min,能量密度为204J/cm~2;以波长630nm的半导体激光照射HpD-PDT组肿瘤,功率密度200mW/cm~2,每光斑照射20min,能量密度为240J/cm~2。于PDT后9d处死鼠行酶联免疫吸附法(ELISA)的双抗体夹心法检测HPPH-PDT和HpD-PDT后突变型p53蛋白(mutant type p53 protein,mtp53)和p16蛋白表达变化。结果:空白、单注药和单照光三组对照组之间mtp53蛋白和p16蛋白表达值两两比较,差异无统计学意义(P>0.05)。HPPH-PDT各剂量组与空白对照组比较,mtp53蛋白表达值明显降低,p16蛋白表达值明显升高,差异有统计学意义(P<0.01)。0.3mg/kg组和0.45mg/kg组与0.15mg/kg组相比,mtp53蛋白表达值降低,p16蛋白表达值升高,差异有统计学意义(P<0.05),0.45mg/kg组mtp53蛋白表达值稍低于0.3mg/kg组,p16蛋白表达值稍高于0.3mg/kg组,差异无统计学意义(P>0.05)。HPPH-PDT0.3mg/kg组和0.45mg/kg组的mtp53蛋白表达值低于HpD-PDT组,p16蛋白表达值高于HpD-PDT组,差异有统计学意义(P<0.05)。结论:HPPH-PDT0.3mg/kg是适宜的HPPH剂量。HPPH-PDT诱导mtp53蛋白表达降低,p16蛋白表达升高。与HpD-PDT相比,HPPH-PDTmtp53蛋白表达更低,p16蛋白表达更高。
Objective: To investigate the effect of HPPH photodynamic therapy on the expression of mutant p53 and p16 proteins in murine G422 glioma and to compare with HpD. Methods: The mouse model of G422 glioma subcutaneously transplanted was established. A blank control group was established. The rats in the control group were injected with HPPH (0.45mg / kg), single 665nm laser irradiation, HPPH-PDT group (0.15mg / kg group, 0.45 mg / kg group) and HpD-PDT group (5 mg / kg). Single injection of HPPH group, HPPH-PDT group and HpD-PDT group were injected into the tail vein with photosensitizer. After 24 h, the HPPH-PDT group and the single laser group were irradiated by the semiconductor laser with the wavelength of 665nm. The power density was 200mW / cm2. The energy density was 204J / cm ~ 2 with irradiation of light beam for 17min, and the tumors of HpD-PDT group were irradiated with semiconductor laser with wavelength of 630nm. The power density was 200mW / cm ~ 2 and the energy density was 240J / cm2. The expression of mutant type p53 protein (mtp53) and p16 protein in HPPH-PDT and HpD-PDT was detected by double-antibody sandwich enzyme-linked immunosorbent assay (ELISA) Results: There was no significant difference in the expression of mtp53 protein and p16 protein between blank control group, single injection group and control group (P> 0.05). Compared with the blank control group, the expression of mtp53 protein in HPPH-PDT groups was significantly decreased and the expression of p16 protein was significantly increased (P <0.01). The protein expression of mtp53 decreased and the expression of p16 protein increased in 0.3mg / kg and 0.45mg / kg groups compared with 0.15mg / kg group, the difference was statistically significant (P <0.05), and mtp53 Protein expression was slightly lower than 0.3mg / kg group, p16 protein expression slightly higher than the 0.3mg / kg group, the difference was not statistically significant (P> 0.05). The expression of mtp53 protein in HPPH-PDT 0.3 mg / kg group and 0.45 mg / kg group was lower than that in HpD-PDT group, and the expression of p16 protein was higher than that in HpD-PDT group (P <0.05). Conclusion: HPPH-PDT 0.3 mg / kg is the appropriate dose of HPPH. HPPH-PDT induced mtp53 protein expression decreased, p16 protein expression increased. Compared with HpD-PDT, HPPH-PDTmtp53 protein expression is lower, p16 protein expression is higher.