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目的观察经5-氨基酮戊酸光动力疗法(5-aminolevulinic acid-mediated photodynamic therapy,ALA-PDT)处理后的食管癌Eca-109细胞,其凋亡相关基因Bcl-xL,Bad和Bid的变化,初步探讨AL-PDT对凋亡通路的作用。方法实验分为两组,分别为对照组和ALA-PDT组,治疗组细胞给予ALA-PDT处理,ALA终浓度为0.25 mM,孵育6 h后,采用波长630 nm激光照射,激光功率200 mW/cm~2,照光150 s,能量密度301/cm~2。对照组仅给予相同体积10%FBS-1640培养基孵育6 h,不加入光敏剂和照光处理。两组细胞于处理后继续于37℃,5%CO_2培养箱内分别培养0.5、3和24h后收集细胞,提取细胞RNA,采用PCR法检测Bcl-xL,Bad和Bid基因表达的变化。结果 Bcl-xL在ALA-PDT作用于Eca-109细胞后3和24 h其表达均显著下调(P均<0.05),Bad在作用后3和24 h,Bid在作用后0.5,3和24 h表达与对照组相比均轻微上调,但与对照组相比其差异无统计学意义(P均>0.05)。结论 ALA-PDT作用于食管癌细胞,可能激活了线粒体凋亡通路,Bcl-xL可能为其作用基因。
Objective To investigate the changes of apoptosis-related genes Bcl-xL, Bad and Bid in Eca-109 cells treated with 5-aminolevulinic acid-mediated photodynamic therapy (ALA-PDT) To investigate the effect of AL-PDT on apoptosis pathway. Methods The experiment was divided into two groups: control group and ALA-PDT group. ALA-PDT treatment group was treated with ALA-PDT. The final concentration of ALA was 0.25 mM. After incubation for 6 h, the cells were irradiated by 630 nm laser with a laser power of 200 mW / cm ~ 2, light 150 s, energy density 301 / cm ~ 2. The control group was given only the same volume of 10% FBS-1640 medium for 6 h, without adding photosensitizer and light treatment. After treatment, the two groups of cells were cultured in 5% CO 2 incubator at 37 ℃ for 0.5, 3, and 24h respectively. The cells were collected and RNA was extracted. The expression of Bcl-xL, Bad and Bid genes were detected by PCR. Results The expression of Bcl-xL was significantly down-regulated in Eca-109 cells at 3 and 24 h after ALA-PDT treatment (all P <0.05). After 3 and 24 h, Compared with the control group, the expression was slightly upregulated, but there was no significant difference between the two groups (P> 0.05). Conclusion ALA-PDT may activate mitochondrial apoptosis pathway in esophageal cancer cells, and Bcl-xL may be the gene of its role.