Effective component from verbena officinalis L.inhibits proliferation and induces apoptosis of human choriocarcinoma JAR cells

Objective:To examine the action of the effective component,4'-methylether-scutellarein,from Verbena officinalis L.(VOL)on the proliferation and apoptosis of human choriocarcinomaJAR cells.Methods:Cell proliferation was measured by MTT[3-(4,5-dimethylthiazol-2-yl)-2,5-di-phenyl tetrasodium bromide,MTT]assay and the incorporation of tritiated thymidine(~3H-TdR).Apoptosis of cell was evaluated by flow cytometry(FCM)and the characteristic apoptoticDNA ladder by agarose gel electrophoresis,and the morphological changes of apoptotic JAR cellswere observed under fluorescence microscopy and electron microscopy(EM).Expressions of ap-optosis proteins,poly(ADP-ribose)polymerase(PARP)and caspase-3,-8,and -9 were deter-mined with Western blot.Results:The effective component from VOL inhibited the proliferation of JAR cells in a dose-and time-dependent manner.The treated cell cycle was arrested in S phase and an apoptotic peakwas found in S phase using FCM analysis.A typical DNA ladder appeared in the treatment groupwhen analyzed by agarose gel electrophoresis.Using fluorescence microscopy,the percentage ofapoptotic cell was 0.9%,6%,and 14% after treatments of 10,20,and 40 mg·L^(-1) of the effec-tive component,respectively,for 48 h.Typical apoptotic changes,such as condensed chromatinand presence of apoptotic bodies,were observed under EM.Treatment with effective componentfor 48 h and 72 h also induced protein expression of PARP and caspase-3,-8,and -9 as seen byWestern blot.Conclusions:The effective component from VOL inhibits cell proliferation and induces apop-tosis in human choriocarcinoma JAR cells.

Objective： To examine the action of the effective component, 4＇-methylether -scutellarein, from Verbena officinalis L. （VOL） on the proliferation and apoptosis of human choriocarcinoma JAR cells. Methods： Cell proliferation was measured by MTT [3-（4, 5-dimethylthiazol-2-yl） -2, 5-di- phenyl tetrasodium bromide, MTT] assay and the incorporation of tritiated thymidine （3H- TdR）. Apoptosis of cell was evaluated by flow cytometry （FCM） and the characteristic apoptotic DNA ladder by agarose gel electrophoresis, and the morphological changes of apoptotic JAR cells were observed under fluorescence microscopy and electron microscopy （EM）. Expressions of apoptosis proteins, poly （ADP-ribose） polymerase mined with Western blot. （PARP） and caspase-3, -8, and -9 were determined with Western blot. Results. The effective component from VOL inhibited the proliferation of JAR cells in a doseand time-dependent manner. The treated cell cycle was arrested in S phase and an apoptotic peak was found in S phase using FCM analysis. A typical DNA ladder appeared in the treatment group when analyzed by agarose gel electrophoresis. Using fluorescence microscopy, the percentage of apoptotic cell was 0.9%, 6%, and 14% after treatments of 10, 20, and 40 mg · L^-1 of the effective component, respectively, for 48 h. Typical apoptotic changes, such as condensed chromatin and presence of apoptotic bodies, were observed under EM. Treatment with effective component for 48 h and 72 h also induced protein expression of PARP and caspase-3, -8, and -9 as seen by Western blot. Conclusions： The effective component from VOL inhibits cell proliferation and induces apoptosis in human choriocarcinoma JAR cells.