核黄素代谢途径影响顺铂对卵巢癌黏附迁移作用研究

Effects of Riboflavin Metabolic Pathways Combination with Cisplatin on Adhesion and Migration of Ovarian Cancer cells

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 investigate the effects of interference of riboflavin （RIB） metabolic pathways on adhesion and migra- tion to cisplatin （DDP） in ovarian cancer HO8910 cells. METHODS Intervention RIB metabolic pathways by lcntiviral vector har- boring 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, RFF2 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 West- ern blot, and the expressions of TNF-a, NF-KB/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-aand NF-KB/p65. However, the RIB could weaken the effects of DDP on HO8910 cell. CONCLUSION Interference metabolic pathway of RIB can enhance DDP effects on ad- hesion and migration viabilities of ovarian cancer HO8910 cell lines, and the effects are associated with blocking the pathway of TNF- a/NF-KB; However, RIB could attenuate the anti-tumor effects of cisplatin on HO8910 cell.