光动力治疗联合顺铂对食管癌Eca-109细胞增殖、凋亡及p38 MAPK通路的影响  

Effects of photodynamic therapy combined with cisplatin on proliferation,apoptosis of esophageal cancer Eca-109 cells and p38 MAPK pathway

作　　者：张明[1] 赵静[1] 李珊[1] 段昕波[1] 吕品田[1] 郑振茹[1] 杜东杰[1] 刘斌[1] ZHANG Ming;ZHAO Jing;LI Shan;DUAN Xin-bo;LYU Pin-tian;ZHENG Zhen-ru;DU Dong-jie;LIU Bin(Department of Oncology,Hebei General Hospital,Shijiazhuang,Hebei 050051,China)

机构地区：[1]河北省人民医院肿瘤科,河北石家庄050051

出　　处：《中国临床研究》2019年第1期13-17,22,共6页Chinese Journal of Clinical Research

基　　金：河北省科技计划项目(15277728D)~~

摘　　要：目的探讨光动力治疗(PDT)联合顺铂(DDP)对食管癌Eca-109细胞增殖、凋亡的影响,以及与p38信号通路的关系,以期对食管癌的治疗提供一定的参考。方法体外培养食管癌Eca-109细胞,分为对照组、PDT组(添加光敏剂5-氨基酮戊酸(5-ALA)后采用630 nm波长、10 J/cm2密度激光照射细胞处理)、DDP组(细胞培养液中添加20 mg/L顺铂处理)、联合组(PDT联合DDP处理)。CCK-8法检测细胞增殖情况;平板细胞克隆实验检测细胞集落形成情况; Hoechst33342染色法观察细胞形态变化; Tunel法、PI/Annexin V-FITC双染法检测细胞凋亡情况;WB法检测细胞中磷酸化-丝裂原活化蛋白激酶p38(p-p38MAPK)、B淋巴细胞瘤-2蛋白(Bcl-2)、Bcl-2相关X蛋白(Bax)、Survivin蛋白表达情况。结果对照组细胞形态、大小均正常; PDT组、DDP组细胞体积明显变小,细胞核荧光强度增高,部分出现碎片化;联合组细胞核固缩现象更加明显。与对照组相比,其他三组细胞抑制率、凋亡率、p-p38MAPK、Bax蛋白表达升高(P均<0. 05);细胞集落数、Bcl-2、Survivin蛋白表达降低(P均<0. 05)。与PDT组、DDP组相比,联合组细胞抑制率、凋亡率、p-p38MAPK、Bax蛋白表达升高(P均<0. 05);细胞集落数、Bcl-2、Survivin蛋白表达降低(P均<0. 05)。结论 PDT联合DDP可能通过抑制p38 MAPK信号通路的激活,进而抑制食管癌Eca-109细胞增殖,诱导其凋亡,发挥肿瘤抑制作用。Objective To explore the effect of photodynamic therapy( PDT) combined with cisplatin treatment on the proliferation and apoptosis of esophageal cancer Eca-109 cells and its association with p38 signaling pathway to provide some reference for the treatment of esophageal cancer. Methods Eca-109 cells were cultured in vitro and were divided into blank control group,PDT group( Eca-109 cells were irradiated by laser with 630 nm wavelength and 10 J/cm^2 density after adding 5-ALA),DDP group( adding 20 mg/L cisplatin to cell culture medium) and combined therapy group( treatment with PDT and DDP). CCK-8 was used to detect cell proliferation. Cell plate colony forming assay was used to detect cell colony formation. Hoechst33342 staining method was used to observe cell morphological changes. TUNEL method and PI/Annexin V-FITC double staining method were used to detect cell apoptosis. Western blot method was used to detect the expressions of phosphorylated p38 mitogen activated proteinase( p-p38 MAPK),B-cell lymphoma-2( Bcl-2),Bcl-2 associated X protein( Bax) and Survivin proteins. Results In blank control group,the cell morphology and size were normal. In PDT group and DDP group,the cell volume obviously diminished,and the nucleus showed enhanced fluorescence intensity and partial fragmentation. In combined therapy group,nuclear condensation was more obvious. Compared with blank control group,cell inhibition rate,cell apoptosis rate and the expression levels of p-p38 MAPK and Bax proteins significantly increased,while the number of cell colony,expression levels of Bcl-2 and Survivin proteins significantly decreased in the other three groups( all P < 0. 05). Compared with PDT group and DDP group,cell inhibition rate,cell apoptosis rate,expression levels of p-p38 MAPK and Bax proteins significantly increased( all P < 0. 05),while the number of cell colony,expression levels of Bcl-2 and Survivin proteins significantly decreased in combined therapy group( all P <0. 05). Conclusion PDT combined with DDP therapy plays an anti-t

关 键 词：食管癌 光动力治疗法 顺铂 增殖 凋亡 丝裂原活化蛋白激酶p38信号通路 

分 类 号：R735.1[医药卫生—肿瘤]