目的 体外研究核黄素(RIB)代谢途径对顺铂(DDP)对卵巢癌HO8910细胞株黏附和迁移的作用。方法 通过慢病毒包装核黄素转运体2(RFT2)shRNA干扰载体和RIB竞争性抑制剂氯丙嗪(CHL)干预RIB代谢途径。HO8910卵巢癌细胞随机分为空白对照组、shRNA对照组、RFT2 shRNA组、CHL(50 μmol·L-1)组、DDP(20 μmol·L-1)组、RFT2 shRNA+DDP组、CHL+DDP组和DDP+RIB(20 μmol·L-1)组。各组细胞处理48 h后,细胞黏附实验检测细胞黏附能力;Transwell方法检测细胞迁徙能力;Western blot检测VEGF、MMP9和MMP2的表达;激光共聚焦检测TNF-α和NF-κB/p65的表达。结果 与单独给药组对比,RFT2 shRNA或CHL联合DDP能够明显降低HO8910细胞黏附和迁移能力;降低MMP9和MMP2的表达;减少细胞TNF-α、NF-κB/p65的表达,RIB可以减弱DDP的作用。结论 干扰FIB代谢途径能够增强DDP对卵巢癌HO8910细胞黏附和迁移能力的作用,该作用与抑制TNF-α/NF-κB途径有关。
Abstract
OBJECTIVE To investigate the effects of interference of riboflavin (RIB) metabolic pathways on adhesion and migration to cisplatin (DDP) in ovarian cancer HO8910 cells. METHODS Intervention RIB metabolic pathways by lentiviral vector harboring shRNA of riboflavin transporter 2 (RFT2) and chlorpromazine (CHL), a competitive inhibitor of RIB. HO8910 ovarian cancer cell line was divided into normal control group, shRNA control group, RFT2 shRNA group, CHL (50 μmol·L-1) group, DDP (20 μmol·L-1) group, RFT2 shRNA+DDP group, CHL + DDP group and DDP + RIB (20 μmol·L-1) group. Each group cells were collected after treatment 48 h according to the design. And then cell adhesive abilities were detected by adhesion experiment, the cells invasive abilities were observed by transwell method, the protein expressions of VEGF, MMP9 and MMP2 were detected by Western blot, and the expressions of TNF-α, NF-κB/p65 were assayed with laser confocal microscopy. RESULTS Compared with the sole DDP treatment group, RFT2 shRNA or CHL combination with DDP had great advantages in reducing cell adhesion and migration viabilities, deceasing expressions of MM9 and MMP2, reducing cell expressions of TNF-α and NF-κB/p65. However, the RIB could weaken the effects of DDP on HO8910 cell. CONCLUSION Interference metabolic pathway of RIB can enhance DDP effects on adhesion and migration viabilities of ovarian cancer HO8910 cell lines, and the effects are associated with blocking the pathway of TNF-α/NF-κB; However, RIB could attenuate the anti-tumor effects of cisplatin on HO8910 cell.
关键词
核黄素 /
氯丙嗪 /
顺铂 /
卵巢癌 /
迁移
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Key words
riboflavin /
chlorpromazine /
cisplatin /
ovarian cancer /
migration
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中图分类号：
R965
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参考文献
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脚注
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基金
浙江省医药卫生科学研究基金资助项目(2013KYA236);丽水学院博士科研启动基金资助(QD1229)
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