ALA联合HPD光动力学治疗鼠G422脑胶质细胞肿瘤基因P16及P53变化的研究

Research of ALA Combined with HPD-PDT Which Induced Change of P16 Gene and P53 Gene of G422 Glioma of Rats

目的:研究ALA、HPD及ALA联合HPD光动力学治疗鼠G422脑胶质细胞瘤肿瘤基因P16及P53表达的变化。方法:以不同剂量ALA(20mg/kg4、0 mg/kg、60 mg/kg、80 mg/kg1、60 mg/kg)、HPD(1 mg/kg、2 mg/kg、3 mg/kg、4 mg/kg、5mg/kg)及不同剂量组合ALA联合HPD光动力学治疗鼠G422脑胶质细胞肿瘤以波长630nm半导体激光照射,功率密度200mW/cm2,每光斑照射20分钟,PDT后一天、三天、七天及十四天,以流式细胞仪测试各组基因P16、P53不同光敏剂组及不同时间表达的变化并与不照光对照组比较。结果:本实验中未作PDT治疗的对照组肿瘤,肿瘤生长快,P16对照组最低值12.01%.最高值15.41%,HPD-PDT组表达最低值17.12%,最高值34.735%,ALA-PDT组,表达最低值17.1%,最高值25.51%,而HPD与ALA联合治疗组表达最低值16.5%,最高值28.22%;P53对照组最低值15.02%,最高值23.16%,HPD-PDT组,表达最低值29.98%,最高值43.62%,ALA-PDT组,表达最低值25.06%,最高值33.25%,而HPD与ALA联合治疗组表达最低值28.08%,最高值41.01%。总的P16、P53光动力学治疗组表达明显高于对照组,随光敏剂剂量增加表达值渐增,HPD-PDT组表达高于ALA组,HPD1mg/kg联合、ALA60-80mg/kg,HPD2mg/kg联合ALA40-60mg/kg,HPD3-4mg/kg联合ALA20mg/kg能达到P16表达最佳值及最高值。PDT后七天表达较佳。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HPD1mg/kg联合ALA60-80mg/kg,HPD2mg/kg联合ALA40-60mg/kg光动力学治疗表达是最佳值或最高值,这样可明显提高ALA-PDT疗效及减少HPD-PDT的皮肤光毒反应。

Objective：Research of ALA, HPD, ALA combined with HPD-PDT which induced P16 gene or P53 gene of G422 glioma of rats. Methods： G422 glioma cells of Rat were randomly divided into several groups and incubated with ALA（20 ,40, 60 ,80,160 mg/kg）,HPD（1,2,3,4,5 mg/kg）andtheir combination dosages. 630nm light（total output 2W） was delivered to tumor at a constant fluence rate： 200mW/cm^2 and a constant irradiated time period： 20 minutes. We set 3 groups （no photo- sensitizers or no irradiation or neither） to be the control groups. We used flow cytometry technology to determine numerical value of P16 gene or P53 gene of G422 glioma tumor of rats during the after PDT 1,3,7,14 day. Results： The growths were rapidness of tumor of groups of no photosensitizers or no irradiation or neither. The rare P16 gene were 12.01%-15.41%, moreover, the HPI〉PDT groups were17.12%-34.735%, he ALA PDT groups were17.1%-25.51% ,thetwo-photsensitizer combination groups were 16.5%-28.22%;The P53 gene were 15.02%-23.16%, moreover, the HPD-PDT groups were 29.98%-43.62%, The ALA PDT groups were 25.06%-33.25%,the two-photsensitizer combination groups were 28.08%-41.01%. Total The rare P16 and P53 gene increased as PDT groups more Non PDT groups. The HPD-PDT groups more ALA-PDT groups, more photosensitizers were added into it. Two-photsensitizer combination groups, HPD 1mg/kg, combining ALA60-80mg/kg, HPD 2mg/kg, combining ALA40-60mg/kg, can reach the optimal and highness level. The optimal expression is 7 day. Conclusion： Suppose PDT can activation function of P16 gene and p53 gene to indueed cells apoptosis or death. The combination dosage of HPD 1mg/kg combination ALA 60-80 mg/kg, and HPD2 mg/kg combination ALA 40-60 mg/kg PDT can obtain curative effect of the HPD5 mg/kg PDT. Therefore it can obvious increase curative effect of ALA-PDT and can reduce cutaneous photic toxicity reaction of HPD-PDT.