The Effect Of Valjatrate E On The Proliferation, Invasion And Migration Of HepG2

Object To investigate the effect of Valjatrate E, a iridoid isolated from Valeriana jatamansi, to block the progression of migration, invasion and proliferation of HepG2. To further study in the role of Valjatrate E"s in MAPK/ERK signaling pathway and to illustrate its mechanism.Methods 1. To evaluate the influence of different concentrations of Valjatrate E(12、10、8、 6、5、4μg/mL) and different time(1、12、24、48hr) on HepG2 cell viability which were measured by MTT.2. Experiments of Zymography, wound healing assay and Transwell were performed to assess invasion ability to HepG2.3. Homogeneity and heterotypic adhesion experiments were utilized to evaluate the adhesion characters of this tumor cell lines, namely, the HepG2.4. Western Blot was tested to evaluate the expression of MMP-2、ERK1/ERK2、 p-ERK1/ERK2, which were considered to being responsible to the development of tumor.Results 1. MTT assay showed Valjatrate E could inhibited significantly the proliferation of HepG2 cells in both time-dependent and dose-dependent manners. The survival rate was tested as 88.71%、82.42%、53.16%、48.25% for Valj atrate E(12μg/mL) at 1h、12h、24h、 48h.2. Wound healing assay showed that gap rate was 51.97%、87.39%、76.34%、57.04% for Valjatrate E(0μg/mL、6μg/mL,3μg/mL、1.5μg/mL) at 48h. Transwell assay showed that invasion(migration) ratio were 100.00%(100.00%)、9.71%(21.97%)、52.39%(57.67%)、 95.39%(97.28%) for Valjatrate E(0μg/mL、12μg/mL、6μg/mL、3μg/mL) at 24h respectively. Zymography experiments show that the expression rate of MMP-2(MMP-9) respectively: 100.00%(100.00%)、13.56%(18.84%)、42.33%(34.06%)、39.07%(33.82%) after HepG2 cells which were affected by Valjatrate E(0μg/mL、12μg/mL、6μg/mL、3μg/mL) for 24h. The three experiments all had the tendency in dose-dependent.3. Homogeneity adhesion assay shows the adhesion rates (No adhesion rates) respectively: 100.00%(100.00%).97.68%(103.12%),99.00%(103.31%),99.19%(93.35%) and heterotypic adhesion assay shows the adhesion rates respectively:100%,67.80%,76.18%,96.54% after HepG2 cells were affected by Valjatrate E(0μg/mL、12μg/mL、6μg/mL、3μg/mL) for 1h.4. Western Blot showed the expression rate MMP-2、ERK1/ERK2、p-ERKl/ERK2 and activated rate ERK1/2 respectively:MMP-2(100.00%、63.17%、105.45%、100.43%)、 ERK1/ERK2(100.00%/100.00%、95.22%/97.73%、92.18%/99.02%、94.79%/99.96%)、 p-ERK1/ERK2(100.00%/100.00%、70.54%/85.95%、85.28%/88.52%、101.31%94.48%)、 ERK1 and (2)activated rate(28.10%(32.29%)、20.82%(28.40%)、26.00%(28.87%)、 30.03%(30.52%)) after HepG2 cells were affected by Valjatrate E(0μg/mL、12μg/mL、 6μg/mL、3μg/mL) for 24h.Conclusions The results show that Valjatrate E can effectively inhibit the ability of migration, invasion and proliferation of HepG2 cells. The effect may relate with the modulatory action on MAPK/ERK signaling pathway.