College of Public Health, Xinjiang Medical University, No 567, SHangde North Road, SHuimogou District, Urumqi, Xinjiang, China.
The First People's Hospital of Urumqi, Urumqi, Xinjiang, China.
Photodiagnosis Photodyn Ther. 2021 Dec;36:102605. doi: 10.1016/j.pdpdt.2021.102605. Epub 2021 Oct 27.
To evaluate the therapeutic effect of Chlorin e6 photodynamic therapy (Ce6-PDT) in human colorectal cancer cells and investigate the role of autophagy in Ce6-PDT.
SW480 cells underwent Ce6-PDT with and without pretreatment with the autophagy inhibitor 3-methyladenine (3MA). Cell viability was assessed using 3-(4,5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assay. Apoptosis was evaluated using an Annexin V assay, using a rhodamine 123 (RH123) assay to evaluate mitochondrial membrane potential (MMP), and by measuring Caspase-3 and Bcl-2 protein expression using western blotting. Autophagy was evaluated by directly visualizing acridine orange-stained acidic vesicular organelles (AVOs) using fluorescent microscopy and by measuring LC3Ⅰ/Ⅱand Atg5 expression using western blotting.
Ce6-PDT decreased SW480 viability in a dose-dependent manner. Ce6-PDT induced apoptosis in SW480 cells via the mitochondrial apoptosis pathway as indicated by decreased mitochondrial membrane potential, increased Annexin V staining, and increased Caspase-3 expression. Ce6-PDT was also shown to induce autophagy as demonstrated by increased acridine-orange stained AVOs as well as increased expression of the autophagy-associated proteins Atg5. Inhibition of autophagy with 3MA potentiated SW480 cell response to Ce6-PDT and increased the rate of apoptosis in the treated cells.
Ce6-PDT induces autophagy and apoptosis of SW480 cells in a dose-dependent manner. Inhibition of autophagy increases the apoptosis induced by Ce6-PDT. Modulation of autophagy may be a potential therapeutic target for colon cancer cells treated with Ce6-PDT.
评估氯乙啶 6 光动力疗法（Ce6-PDT）在人结肠癌细胞中的治疗效果，并研究自噬在 Ce6-PDT 中的作用。
SW480 细胞接受 Ce6-PDT 治疗，并在预处理时加入自噬抑制剂 3-甲基腺嘌呤（3MA）。采用 3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四唑溴盐（MTT）测定细胞活力。采用 Annexin V 测定法评估细胞凋亡，采用罗丹明 123（RH123）测定法评估线粒体膜电位（MMP），并通过 Western blot 测定 Caspase-3 和 Bcl-2 蛋白表达。通过荧光显微镜直接观察吖啶橙染色的酸性囊泡细胞器（AVOs）和 Western blot 测定 LC3Ⅰ/Ⅱ和 Atg5 表达来评估自噬。
Ce6-PDT 以剂量依赖性方式降低 SW480 活力。Ce6-PDT 通过线粒体凋亡途径诱导 SW480 细胞凋亡，表现为线粒体膜电位降低、Annexin V 染色增加和 Caspase-3 表达增加。Ce6-PDT 还诱导自噬，表现为吖啶橙染色的 AVOs 增加以及自噬相关蛋白 Atg5 的表达增加。用 3MA 抑制自噬增强了 SW480 细胞对 Ce6-PDT 的反应，并增加了处理细胞中的凋亡率。
Ce6-PDT 以剂量依赖性方式诱导 SW480 细胞自噬和凋亡。抑制自噬增加了 Ce6-PDT 诱导的凋亡。调节自噬可能是用 Ce6-PDT 治疗结肠癌细胞的潜在治疗靶点。
