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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(50μmol·L~(-1))组、DDP(20μmol·L~(-1))组、RFT2 shRNA+DDP组、CHL+DDP组和DDP+RIB(20μmol·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。
Objective To study the effect of riboflavin (RIB) metabolic pathway on the adhesion and migration of ovarian cancer cell line HO8910 by cisplatin (DDP) in vitro. Methods Interfering RIB metabolic pathway by lentivirus packaging riboflavin transporter 2 (RFT2) shRNA interference vector and RIB competitive inhibitor chlorpromazine (CHL). HO8910 ovarian cancer cells were randomly divided into blank control group, shRNA control group, RFT2 shRNA group, CHL (50μmol·L -1) group, DDP (20μmol·L -1) group, RFT2 shRNA + DDP group , CHL + DDP group and DDP + RIB group (20μmol·L -1). Cell adhesion assay was used to detect cell adhesion after 48 h of treatment. Cell migration was evaluated by Transwell assay. Expressions of VEGF, MMP9 and MMP2 were detected by Western blot. Expressions of TNF-α and NF-κB / p65 were detected by confocal laser scanning microscopy. Results Compared with the single administration group, RFT2 shRNA or CHL combined with DDP could significantly reduce the adhesion and migration ability of HO8910 cells; reduce the expression of MMP9 and MMP2; decrease the expression of TNF-α and NF-κB / p65; effect. Conclusion The interference of FIB metabolic pathway can enhance the effect of DDP on the adhesion and migration of ovarian cancer HO8910 cells, which is related to the inhibition of TNF-α / NF-κB pathway.