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.