5-ALA-PDT法在HPV11.HaCaT细胞模型中的应用

The application of 5-ALA-PDT in HPV11.HaCaT cell model

目的:确定光动力疗法(5-ALA-PDT)对携带HPV11全基因组的HaCaT细胞(HPV11.HaCaT)模型的影响。方法:HPV11.HaCaT细胞培养传代后分别用不同浓度5-ALA(0.375、0.75、1.5、3 mM),或不同剂量红光(10、20、40 J/cm^2),或不同浓度5-ALA结合不同剂量红光处理。用MTT法检测细胞增殖,实时荧光定量PCR测定HPV11早期基因E5、E6和E7 mRNA的表达。结果:0.75mM以上浓度的5-ALA联合20或40 J/cm^2红光,细胞存活率均在50%以下(P<0.01)。0.375 mM 5-ALA联合20 J/cm^2红光能明显降低HPV11 E6和E7 mRNA表达,相对表达量为0.28和0.26(P<0.01),而0.75 mM 5-ALA联合20 J/cm^2红光处理对HPV11 E6和E7 mRNA表达影响不明显,相对表达量为0.85和1.46(P>0.05),但以上两种处理条件对HPV11 E5 mRNA的表达均无明显影响。结论:5-ALA-PDT法应用于HPV11.HaC aT细胞模型能抑制HPV11.HaC aT细胞增殖,并能降低该细胞中HPV11 E6、E7mRNA的表达。

Objective：To determine the impact of 5-ALA-PDT on the HPV11.HaCaT cell model （HaCaT keratinocytes harboring human papillomavirus type 11 episome ） . Methods： Cultured HPV11. HaCaT cells were treated with 5-ALA （0.375、0.75、1.5、3 mM）, red light irradiation （10、20、40 J/cm^2）, or different concentration of 5-ALA combined with different dose of red light irradiation. The effects on cells proliferation were examined by MTT assay. The alterations of HPV11 E5, E6 and E7 mRNA expression level were exam-ined by real-time fluorescence quantitative PCR. Results：When 5-ALA concentration was higher than 0.75 mM and combined with 20 or 40 J/cm^2 red light irradiation, the survival rate of the cells was less than 50%（ P〈0.01） . When 0.375 mM 5-ALA combined with 20 J/cm^2 red light irradiation, the expression of HaCaT keratinocytes was significantly reduced, with relative expression rates of 0.28 and 0.26 （ P〈0.01） . 0.75 mM 5-ALA combined with 20 J/cm^2 red light irradiation did not inhibit the expression of HPV11 E6 and E7 mR-NA, with relative expression rates of 0.85 and 1.46 （P〉0.05）. However,the two kinds of treatment mentioned above did not affect the expression of HPV11 E5 mRNA. Conclusion：5-ALA-PDT can effectively impact on the HPV11. HaCaT cell model, such as the proliferation of the HPV11. HaCaT cells and the expression of HPV11 E6 and E7 mRNA.