Luo Mengyu, Ji Jiayin, Yang Kaizhen, Li Hongxia, Kang Ling
College of Public Health, Xinjiang Medical University, No 567, SHangde North Road, SHuimogou District, Urumqi, Xinjiang, China; Key Laboratory of Special Environment and Health Research in Xinjiang, China.
The First People's Hospital of Urumqi, Urumqi, Xinjiang, China.
Photodiagnosis Photodyn Ther. 2022 Dec;40:103082. doi: 10.1016/j.pdpdt.2022.103082. Epub 2022 Aug 24.
Photodynamic therapy is a tumour treatment method. Its mechanism mainly induces apoptosis, autophagy, and other ways to cause cell death. Therefore, this study aims to evaluate the therapeutic effect of chlorine e6 photodynamic therapy (Ce6-PDT) combined with oxaliplatin (L-OHP) in colon cancer and to investigate the role of autophagy in L-OHP treatment and Ce6-PDT combined with L-OHP in colon cancer.
CCK-8 assay, Scratch wound healing assay, and Western Blot (WB) were used to identify drug-resistant colon cancer cell line SW620/L-OHP. Annexin V/FITC assay, laser confocal double immunofluorescence staining method and WB were employed to investigate the apoptosis and autophagy changes in Ce6-PDT combined with L-OHP.
Drug resistance cells SW620/L-OHP were developed under the continuous multi-generation of L-OHP treatment, and the expression of ATP-binding cassette subfamily B member 1 (ABCB1) and ATG5 proteins were increased. The results of immunofluorescence showed that LC3B accumulated in SW620 cells and SW620/L-OHP cells under the treatment of L-OHP. The WB results indicated that LC3B and ATG5 protein expression was increasing in SW620 cells and SW620/L-OHP cells. Inhibition of L-OHP-induced autophagy reduces SW620 cells and SW620/L-OHP cells' viability while increasing apoptosis and the Pro Caspase-3 protein expression. The combination of Ce6-PDT and L-OHP decreased the cell viability, the cell migration ability, the Bcl-2 protein expression, and increased the apoptosis rate, Pro Caspase-3 protein expression in SW620 cells.
L-OHP can cause SW620 cells drug resistance. Autophagy plays a protective role in the L-OHP treatment of SW620 cells and SW620/L-OHP cells, and inhibition of autophagy can increase the efficacy of L-OHP. Ce6-PDT combined with L-OHP can further improve the tumor's therapeutic effect, and autophagy inhibition can improve the efficacy of combined therapy.
光动力疗法是一种肿瘤治疗方法。其机制主要通过诱导细胞凋亡、自噬等方式导致细胞死亡。因此，本研究旨在评估氯e6光动力疗法（Ce6-PDT）联合奥沙利铂（L-OHP）治疗结肠癌的疗效，并探讨自噬在L-OHP治疗以及Ce6-PDT联合L-OHP治疗结肠癌中的作用。
采用CCK-8法、划痕伤口愈合试验和蛋白质免疫印迹法（WB）鉴定耐奥沙利铂的结肠癌细胞系SW620/L-OHP。采用膜联蛋白V/异硫氰酸荧光素（Annexin V/FITC）法、激光共聚焦双免疫荧光染色法和WB法研究Ce6-PDT联合L-OHP处理后细胞凋亡和自噬的变化。
在连续多代L-OHP处理下建立了耐药细胞SW620/L-OHP，且三磷酸腺苷结合盒亚家族B成员1（ABCB1）和自噬相关蛋白5（ATG5）的表达增加。免疫荧光结果显示，在L-OHP处理下，LC3B在SW620细胞和SW620/L-OHP细胞中积累。WB结果表明，LC3B和ATG5蛋白在SW620细胞和SW620/L-OHP细胞中的表达增加。抑制L-OHP诱导的自噬会降低SW620细胞和SW620/L-OHP细胞的活力，同时增加细胞凋亡和半胱天冬酶原-3（Pro Caspase-3）蛋白的表达。Ce6-PDT与L-OHP联合使用可降低SW620细胞的细胞活力、细胞迁移能力、Bcl-2蛋白表达，并增加细胞凋亡率、Pro Caspase-3蛋白表达。
L-OHP可导致SW620细胞产生耐药性。自噬在L-OHP治疗SW620细胞和SW620/L-OHP细胞中起保护作用，抑制自噬可提高L-OHP的疗效。Ce6-PDT联合L-OHP可进一步提高肿瘤的治疗效果，抑制自噬可提高联合治疗的疗效。
