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目的检测高表达OAZI-1的小鼠黑素瘤B16-F1细胞对抗癌药物多胺类似物CPENSpm敏感性的影响并探讨其作用机制。方法高表达OAZI-1的B16-F1细胞经CPENSpm处理后,MTT法检测细胞增殖;QT-RT-PCR检测细胞内OAZI-1、ODC和OAZ的mRNA表达水平;反相高效液相色谱法检测细胞内多胺含量;化学发光法分析细胞内SMO酶活性。结果高表达OAZI-1显著增加B16-F1细胞对CPENSpm的敏感性,QT-RT-PCR结果显示,OAZI-1高表达增加ODCmRNA但降低OAZ mRNA水平,CPENSpm处理对ODC和OAZ的mRNA水平无进一步影响。与对照细胞相比,CPENSpm处理能更显著性地降低OAZI-1高表达细胞中的多胺含量,同时更强地诱导SMO酶的活性。结论 CPENSpm处理能在更大程度上耗竭OAZI-1高表达的B16-F1细胞中的多胺,由此对该肿瘤细胞显示出更大的细胞毒性。
Objective To investigate the effect of OAZI-1-overexpressing mouse melanoma B16-F1 cells on the sensitivity of anti-cancer drug polyamine analogue CPENSpm and its mechanism of action. Methods The proliferation of B16-F1 cells with OAZI-1 overexpression was determined by MTT assay after treatment with CPENSpm. The mRNA levels of OAZI-1, ODC and OAZ were detected by QT-RT-PCR and by reverse phase high performance liquid chromatography Intracellular polyamine content; Chemiluminescence method analysis intracellular SMO enzyme activity. Results High expression of OAZI-1 significantly increased the sensitivity of B16-F1 cells to CPENSpm. QT-RT-PCR results showed that OAZI-1 overexpression increased ODC mRNA but decreased OAZ mRNA level, while CPENSpm treatment did not further increase mRNA level of ODC and OAZ influences. Compared with control cells, CPENSpm treatment reduced the polyamine content in OAZI-1 highly expressed cells more significantly while inducing SMO enzyme activity more strongly. Conclusions CPENSpm treatment is capable of depleting polyamines in OAZI-1-overexpressed B16-F1 cells to a greater extent, thereby demonstrating greater cytotoxicity to the tumor cells.