Wang Haiping, Li Long, Wang Pan, Wang Xiaobing, Zhang Kun, Liu Quanhong
Key Laboratory of Ministry of Education for Medicinal Plant Resources and Natural Pharmaceutical Chemistry, College of Life Sciences, Shaanxi Normal University, China.
Key Laboratory of Ministry of Education for Medicinal Plant Resources and Natural Pharmaceutical Chemistry, College of Life Sciences, Shaanxi Normal University, China.
Photodiagnosis Photodyn Ther. 2016 Dec;16:1-8. doi: 10.1016/j.pdpdt.2016.07.004. Epub 2016 Aug 5.
Multidrug resistance (MDR) of breast cancer is a major obstacle in chemotherapy of cancer treatments. Recently the anti-tumor effects of Chlorin e6 (Ce6) mediated photodynamic therapy (Ce6-PDT) were reported in skin cancer and hepatoma in vitro. However, its therapeutic potential in killing human breast cancer especially those with MDR and the differences between MCF-7 and MCF-7/ADR after PDT treatment has not been fully investigated.
MTT assay was used to measure cell survival rate of MCF-7 cells and MCF-7/ADR cells. Intracellular reactive oxygen species (ROS) generation was measured by monitoring the fluorescence intensity of dichlorofluorescein (DCF) by flow cytometry. Nuclear morphology changes and DNA damage in both MCF-7 and MCF-7/ADR after Ce6-PDT were analyzed by hochest33342 staining and comet assay. Western blot and monodansylcadaverine (MDC) staining were used to monitor autophagic response in MCF-7/ADR.
Ce6-PDT induced cell viability decrease, intracellular ROS generation, and DNA damage in concentration-dependent and cell-specific manner, and MCF-7 was more sensitive to Ce6-PDT than MCF-7/ADR cells at the same PDT condition. PDT treatment could trigger cell death via apoptosis in MCF-7 cells but autophagic cell death in MCF-7/ADR cells.
These results suggested that MCF-7 was more sensitive to Ce6-PDT than MCF-7/ADR, and PDT treatment could trigger apoptotic response in MCF-7 cells, but stimulate autophagic response in MCF-7/ADR cells.
乳腺癌的多药耐药性(MDR)是癌症化疗中的主要障碍。最近有报道称,二氢卟吩e6(Ce6)介导的光动力疗法(Ce6-PDT)在体外对皮肤癌和肝癌具有抗肿瘤作用。然而,其在杀伤人类乳腺癌尤其是多药耐药乳腺癌方面的治疗潜力,以及光动力疗法(PDT)治疗后MCF-7和MCF-7/ADR细胞之间的差异尚未得到充分研究。
采用MTT法检测MCF-7细胞和MCF-7/ADR细胞的存活率。通过流式细胞术监测二氯荧光素(DCF)的荧光强度,测定细胞内活性氧(ROS)的生成。采用hochest33342染色和彗星试验分析Ce6-PDT处理后MCF-7和MCF-7/ADR细胞的核形态变化和DNA损伤。采用蛋白质免疫印迹法和单丹磺酰尸胺(MDC)染色监测MCF-7/ADR细胞中的自噬反应。
Ce6-PDT以浓度依赖性和细胞特异性方式诱导细胞活力下降、细胞内ROS生成和DNA损伤,在相同的光动力疗法条件下,MCF-7比MCF-7/ADR细胞对Ce6-PDT更敏感。光动力疗法治疗可通过诱导MCF-7细胞凋亡引发细胞死亡,但可诱导MCF-7/ADR细胞发生自噬性细胞死亡。
这些结果表明,MCF-7比MCF-7/ADR对Ce6-PDT更敏感,光动力疗法治疗可诱导MCF-7细胞发生凋亡反应,但可刺激MCF-7/ADR细胞发生自噬反应。