MPPa光动力疗法诱导人肺癌顺铂耐药细胞凋亡

Induction of Apoptosis of Human Cisplatin-resistance Lung Cancer Cells with MPPa-photodynamic Therapy

肺癌死亡率居世界肿瘤首位,5年生存率不足15%,顺铂耐药是死亡率居高不下的重要原因,本文将探讨光敏剂MPPa介导的光动力疗法诱导人非小细胞肺癌顺铂耐药株A549/DDP发生凋亡的现象及其机制。课题组分别给予不同浓度光敏剂MPPa(0、1、2、4、8、16μmol/L)、不同能量光照(0、0.6、1.2、2.4、3.6、4.8J/cm2)处理细胞,CCK-8法检测细胞活性。将细胞分为对照组、MPPa组(2μmol/L MPPa)、光照组(2.4J/cm2光能量密度)和MPPa介导光动力组(PDT组)(2μmol/L MPPa、2.4J/cm2光能量密度)。应用Anne-V/PI双染流式细胞技术检测细胞凋亡;DCFH-DA染色观察细胞内活性氧产生;蛋白质印迹法(Western blot)检测B淋巴细胞瘤-2(Bcl-2)蛋白和Bcl-2相关X蛋白(Bax)的表达。实验结果显示,单纯光照及MPPa对细胞生长无抑制作用;MPPa介导光动力却对人肺癌耐顺铂细胞A549/DDP有显著杀伤作用,其杀伤作用呈明显的剂量效应关系(P<0.05);PDT组发生凋亡率均高于各对照组(P<0.05);DCFH-DA染色发现PDT组细胞内活性氧明显高于各对照组;Western blot检测发现PDT组Bax蛋白表达升高,Bcl-2蛋白表达降低。实验证实MPPa介导的光动力疗法可抑制肺癌顺铂耐药细胞A549/DDP活性,并诱导其凋亡。

Lung cancer is the leading cause of cancer-related deaths worldwide.Despite the development and use of several targeting drugs for lung cancer therapy,the five-year survival rate has remained as low as 15%for the past three decades.Cisplatin-based chemotherapy is considered the first-line therapeutic strategy for lung cancer.However,developments of chemoresistance is a major obstacle for the successful treatment.Therefore,the development of novel therapy against cisplatin-resistance lung cancer is imperative.Photodynamic therapy（PDT）,which is a non-invasive combinatorial therapeutic modality using light,photosensitizer（PS）and oxygen,may provide an unprecedented tool to develop more effective treatments.To provide experimental basis for its application in cisplatin-resistance lung cancer,we will discuss the biological effects of MPPa-photodynamic therapy in human cisplatin-resistance lung cancer cells in this article.Human cisplatin-resistance lung cancer cells A549/DDP were co-cultured with MPPa（0,1,2,4,8,16μmol/L）and exposed to light（0,0.6,1.2,2.4,3.6,4.8J/cm2）,and cell viability was determined with CCK-8assay.Flow cytometry was used to detect apoptosis,DCFH-DA staining was employed to observe reactive oxygen species（ROS）,and Western blot was used to detect the expressions of B-cell lymphoma-2（Bcl-2）protein and Bcl-2associated X protein（Bax）.The proliferation of A549/DDP cells was suppressed by PDT.The apoptotic rate in the PDT group was significantly higher than that in the control,MPPa or light group（P〈0.05）.The level of ROS was increased.The expression of Bax was increased,and that of Bcl-2was decreased.MPPa-photodynamic therapy can significantly suppress cell viability,and induce apoptosis in human cisplatin-resistance lung cancer cells.