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5-氨基酮戊酸介导的光动力疗法联合顺铂对人卵巢癌OVCAR-3细胞的作用研究

Study on the effect of 5-aminolevulinic acid-mediated photodynamic therapy combined with cisplatin on human ovarian cancer OVCAR-3 ​cells.

作者信息

Lv Haoxuan, Suo Yuping, Sun Qian, Fan Jingjing, Wang Qian

机构信息

Department of Obstetrics and Gynecology of Fifth Clinical Medical College, Shanxi Medical University, Taiyuan, China.

Department of Obstetrics and Gynecology, Shanxi Provincial People's Hospital, Taiyuan, China.

出版信息

Adv Med Sci. 2023 Mar;68(1):147-156. doi: 10.1016/j.advms.2023.03.001. Epub 2023 Mar 23.

Abstract

PURPOSE

This article explores the effect of 5-aminolevulinic acid (5-ALA)-mediated photodynamic therapy (PDT) combined with cisplatin (CDDP) on the apoptosis of human ovarian cancer cells and the mechanism of action of the combination therapy.

MATERIALS AND METHODS

Human ovarian cancer OVCAR-3 ​cells were cultured in vitro and divided into 5-ALA/PDT group, CDDP group and combined treatment group (5-ALA/PDT combined with different concentrations of CDDP). After administration of the corresponding drugs, a CCK-8 assay was used to detect the inhibition rate of cell proliferation. After Rhodamine 123 staining, mitochondrial membrane potential changes were observed under fluorescence microscopy. The apoptosis rate and reactive oxygen species (ROS) content were detected by flow cytometry. Western blotting was used to detect protein expression.

RESULTS

The CCK-8 assay showed that CDDP in combination with 5-ALA/PDT significantly enhanced cytotoxicity compared to treatment with CDDP alone and that low doses of CDDP were sufficient to induce these combination effects. The mitochondrial membrane potential in each combination treatment group gradually decreased with increasing CDDP concentration, while the apoptosis rate and reactive oxygen species (ROS) content detected by flow cytometry gradually increased. Western blotting assay showed that the expression of bax, cleaved caspase-9, cleaved caspase-3, and cleaved PARP was increased, while the expression of bcl-2, caspase-9, caspase-3, and PARP was decreased, and the differences were statistically significant (P ​< ​0.05).

CONCLUSIONS

In summary, 5-ALA/PDT combined with CDDP can effectively inhibit cell proliferation and promote apoptosis, and this combination may induce apoptosis by activating the mitochondrial pathway.

摘要

目的

本文探讨5-氨基酮戊酸(5-ALA)介导的光动力疗法(PDT)联合顺铂(CDDP)对人卵巢癌细胞凋亡的影响及联合治疗的作用机制。

材料与方法

将人卵巢癌OVCAR-3细胞体外培养,分为5-ALA/PDT组、CDDP组和联合治疗组(5-ALA/PDT联合不同浓度的CDDP)。给予相应药物后,采用CCK-8法检测细胞增殖抑制率。罗丹明123染色后,在荧光显微镜下观察线粒体膜电位变化。通过流式细胞术检测凋亡率和活性氧(ROS)含量。采用蛋白质印迹法检测蛋白表达。

结果

CCK-8法检测显示,与单独使用CDDP治疗相比,CDDP联合5-ALA/PDT显著增强了细胞毒性,且低剂量的CDDP足以诱导这些联合效应。各联合治疗组的线粒体膜电位随CDDP浓度增加逐渐降低,而流式细胞术检测的凋亡率和活性氧(ROS)含量逐渐增加。蛋白质印迹法检测显示,bax、裂解的caspase-9、裂解的caspase-3和裂解的PARP表达增加,而bcl-2、caspase-9、caspase-3和PARP表达降低,差异具有统计学意义(P<0.05)。

结论

综上所述,5-ALA/PDT联合CDDP可有效抑制细胞增殖并促进凋亡,这种联合可能通过激活线粒体途径诱导凋亡。

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