| Choriocarcinoma is caused by abnormal development and excessive proliferation of gestational trophoblastic cells. The recovery rate of human choriocarcinoma treated by chemotherapy is up to 80%-90%. Due to its high sensitivity to chemotherapy, choriocarcinoma is thought to be a kind of eusemia gynecologic tumor. However, chemotherapy is not so efficacious in some high-risk and drug resistant cases. In addition, the side effects of the chemical drugs may also influence the chemotherapy effect. For this reason, it has been a hot spot to seek for a novel, high effective, but lower toxic anti-choriocarcinoma drug.Previous studies in our laboratory have shown that C-part of Verbena Offcinalis could inhibit the proliferation of JAR cells, block the cell cycle at G2/M phase and induce cell apoptosis.4'-Methyl ether-scutellarein (4'-MS), a flavanoids chemical, is an effective component extracted from Verbena Offcinalis C-part through a repeated silica gel column chromatography. To investigate the anticancer effect of 4'-MS, human choriocarcinoma JAR cell line was employed as an in vitro model in this study. We determined the effect of 4'-MS on cell proliferation in JAR cells and examined its underlying signal transduction mechanism with a combination of cell and molecular biological methods. The present study provides evidence that Verbena Offcinalis is a potential anticancer drug to treat choriocarcinoma.1. Effects of 4'-MS on proliferation and apoptosis in human choriocarcinoma cells in vitro MTT assay demonstrated that the inhibitory effect of 4'-MS on JAR cells growth developed when the concentration was up to 10μg/mL and was in a dose and time dependent manner. The resultes of FCM analysis showed that 4'-MS treatment could induce a substantial apoptotic response in JAR cells and a dramatical increase of the percentage of cell cycle progression in G2/M phase, both of which were enhanced with the raise of 4'-MS. AO/EB double staining disclosed that there were more viable apoptotic cells in 4'-MS treated groups than control group and the number of non-viable apoptotic cells and dead cells increased with dose increasing. Thus, certain concentration of 4'-MS resulted in a decrease in proliferation and an increase in apoptosis in JAR cells.2. The mechanism of anti-proliferative and apoptotic effects of 4'-MS on human choriocarcinoma cells in vitroSurvivin expression in 4'-MS treated cells was decreased both at mRNA and protein levels according to RT-PCR and Western blot results. It had been observed by RT-PCR that the phosphorylated p38, Caspase 3, Caspase 9 and Survivin mRNA expression increased with 4'-MS addition. Western blot analyses showed that protein expression of cytochrome C, phosphorylated p38, Caspase 3 and Caspase 9 were extremely up-regulated by 20μg/mL and 40μg/mL 4'-MS treatment. So, the molecular mechanism by which 4'-MS participate in the inhibition of Human Choriocarcinoma JAR cells growth might be due to the marked down-regulation of Survivin, as well as the direct activation of Bax, cytochrome C, Caspase 3, Caspase 9 and p38 MAPK.In summary, based on the model of primary cultured Human Choriocarcinoma JAR cells, we demonstrated that 4'-MS could efficiently inhibit the proliferation of JAR cells and induce cell apoptosis, either by down-regulation of Survivin, as well as the direct activation of Bax, cytochrome C, Caspase 3, Caspase 9 and p38 MAPK, which have an active effection on endogenous apoptosis signal transduction pathway.
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