Mechanisms Of The Celastrus Orbiculatus Extracts Inhibit The Metastasis In Human Hepatocarcinoma HepG2Cells

Heptocellular carcinoma is one of the most common cancer of digestive tract.. Surgery is commonly the treatment of choice for HCC. However, tumor metastasis after surgery seriously hamper the long-term efficacy of surgical treatment. In recent study, metastasis is a complex process which is made up of several steps, including cell proliferation, migration, invasion, attachment, and so on.Tumor metastasis is regulated by many signaling pathways. Phosphatidylinositol-3-kinase/Protein Kinase B (PI3K/Akt) signaling pathway is an important intracellular signal transduction pathway. It plays important roles in cell apoptosis and survivalby affecting the activity of downstream effecter molecules, and it is closely associated with the development and progression of human tumors. Study also reveals that PI3K/Akt signaling pathway promotes cell migration and invasion may through increased expression and proteolytic activities of Nuclear Factor Kappa B (NF-κB), Matrix Metalloproteinase-2(MMP-2) and Matrix Metalloproteinase-9(MMP-9).The Celastraceae plant Celastrus orbiculatus(Celastrus orbiculatus Thunb, Nansheteng, Celastrus), which is widely distributed in China, has been used as medicine in China for rheumatism diseases, and others. Our previous study found that COE displays effects in antitumor activity. However, the mechanismsunderlying the antitumor activity of COE remainlargey is unknow. In this study, we investigated the anti-metastasis effects of COE in human hepatocar cinoma HepG2cells.Objective:To study the effect and mechanism of COE on proliferation, invasion and migration in HepG2cells. And to further explore the molecular mechanisms underlying.Methods:After HepG2cells were treated with COE at various concentrations (20,40,80,160 and320μg/ml) for24h or48h, the cell proliferation effect was measured by using the MTT assay. Cell morphology was observed under the inverted microscope. The invasion and migration ability of HepG2cells were assayed by transwell assay. To investigate whether COE could inhibit the PI3K/Akt signaling pathway, NF-κB p65, MMP-2and MMP-9, we used Western boltting assay.Results:The HepG2cells treated with COE (20,40,80,160and320μg/mL) at different concentrations for24h exhibited a dose-dependent growth inhibition. The rates of inhibition were11.74±2.95,12.59±3.16,19.59±3.02,51.94±7.30and58.56±11.83, respectively. When HepG2cells was treated with COE at doses of20to320μg/mL for48h, the rates of inhibition were18.57±9.89,27.11±10.12,39.62±7.63,70.80±3.74and77.27±2.91, respectively. Under the microscope, we found that apoptotic state of the cells and the number of the cells was reduced. The concentrations of20to80μg/mL of COE significantly inhibited the invation and migration ability of HepG2cells. The HepG2cells treated with COE at doses20to160μg/mL for24h. We found that p-Akt, NF-κBp65, MMP-2and MMP-9pritein were reduced, whereas there was no significant effect on Akt.Conclusions:COE could significantly inhibit the proliferation, invasive and migration ability of HepG2cells. It might be due to the down-regulation of PI3K/Akt signaling pathway, NF-κBp65, MMP-2and MMP-9.