【摘要】 为探讨马鞭草中4′-甲醚-黄芩素对人绒癌JAR/VP16耐药细胞株的多药耐药逆转作用及可能的逆转机制,采用四甲基偶氮唑盐比色、透射电镜观察、逆转录多聚酶链反应、流式细胞术、基因芯片以及实时定量PCR等方法,检测到4’-甲醚-黄芩素对化疗药物具有协同增效作用,显著逆转耐药细胞对鬼臼乙叉甙(VP16)、甲氨蝶呤(MTX)及更生霉素(KSM)的耐药性,该药物作用后,耐药细胞超微结构呈现凋亡样改变,细胞凋亡率增高并出现明显凋亡峰.此外,耐药细胞中MDR1、MRP1、MRP2、MRP6、AHR、COMT、FGF2等耐药相关基因以及Bcl-2、BFL1、NAIP、p63等凋亡抑制基因表达降低,而Apaf-1、ASC、ATM、Bad、Bak、Bax、BimL等凋亡促进基因表达升高.上述实验结果表明:4′-甲醚-黄芩素对绒毛膜癌耐药细胞具有显著的耐药逆转作用,这种逆转作用可能是通过降低耐药基因表达、促进细胞凋亡实现的.更多还原

【Abstract】 Several methods, including MTT assay, transmission electron-microscope (TEM), RT-PCR, flow cytometry, gene array and real time PCR, were used to investigate the reversal effect and potential mechanism of 4′-methylether-scutellarein from Verbena officinalis L.(VOL) on mutidrug resistance of JAR/VP16 cells. 4′-methylether-scutellarein produced a synergistic effect combining with the chemotherapeutics and reversed the resistant phenotype of JAR/VP16 cells. TEM analysis and flow cytometry confirmed apoptotic cell features, containing earlier apoptotic modifications, increased apoptotic cell percentage and typical apoptotic peak, when JAR/VP16 cells exposed to 4′-methylether-scutellarein. The gene array results showed that expression levels of some multidrug resistant genes, i.e. MDR1、MRP1、MRP2、MRP6、AHR、COMT and FGF2, were decreased. The expression levels of some apoptosis-promoting genes, i.e. Apaf-1、ASC、ATM、Bad、Bak、Bax and BimL, were increased, whereas those of some apoptosis-suppressing genes, i.e. Bcl-2、BFL1、NAIP and p63, were decreased upon 4′-methylether-scutellarein treatment. Sixteen genes were selected to be detected by real time PCR, and the results were identical with those of gene array. Taken together, 4′-methylether-scutellarein can efficiently reverse multidrug resistance in JAR/VP16 cells. The potential mechanism may be via inhibiting the multidrug resistant gene expressions and/or promoting cell apoptosis.更多还原