【摘要】 目的探讨促癌物佛波酯(12-O-tetradecanoylphorbol-13-acetate,TPA),岗田酸(okadaic ac id,OA)和氯化镉(cadm ium chloride,CdC l2)在促进BALB/c 3T3细胞转化过程中对骨桥素基因(OPN)转录表达的影响。方法以N-甲基-N′-硝基-N-亚硝基胍(MNNG)为启动剂,TPA,OA和CdC l2分别为促癌剂,建立BALB/c 3T3细胞转化模型。采用小鼠毒理基因芯片分别检测促癌物处理过程中和转化细胞集落中的基因表达变化,同时采用实时RT-PCR验证部分细胞样本的OPN基因表达。结果基因芯片检测结果显示,3种促癌物在作用过程中均能诱导OPN基因表达升高,其中TPA处理后4 h,7 d和17 d,OA处理后3,7 d和24 d,CdC l2处理后3 d和7 d,OPN基因的表达较对照组增强;同时在促癌物诱导形成的9个转化细胞集落中,有8个转化集落检测到OPN基因表达升高。此外,实时RT-PCR检测也验证了OPN基因的表达升高。结论OPN基因与TPA,OA和CdC l2的促癌作用存在密切联系。更多还原

【Abstract】 Objective To explore the changes of osteopontin （OPN） gene expression induced by tumor promoter of 12-O-tetradecanoylphorbol13-acetate（TPA）,okadaic acid（OA） or cadmium chloride(CdCl₂).Methods The two-stage transformation test of BALB/c 3T3 cells was established with MNNG as initiator,TPA,OA or CdCl₂ as promoter respectively.Mouse toxicology gene chip was used to detect the gene expression of BALB/c 3T3 cells transformed with TPA,OA or CdCl₂ respectively,and the expression of OPN gene was validated by real-time RT-PCR assay.Results TPA,OA or CdCl₂ could increase the transcriptional expression of OPN gene in BALB/c 3T3 cells.The expression of OPN gene was up-regulated in the eight transformed cell colonies induced by TPA,OA or CdCl₂ respectively.Moreover,the up-regulation of OPN gene expression was confirmed by real-time RTPCR assay.Conclusion The up-regulation of OPN gene expression is closely related to cell transformation of BALB/c 3T3 cells induced by TPA,OA or CdCl₂.更多还原