ABSTRACT
Objective To investigate the effect and mechanism of acteoside (ACT) in inhibiting epithelial-mesenchymal transition (EMT) in human hepatoma HCCLM3 cells by regulating the ERK1/2 pathway. Methods CCK-8 assay was used to detect the effect of hepatocellular carcinoma cell proliferation. The invasion and migration of HCC cells were detected by scratch and Transwell tests. The mRNA and protein expression levels of the ERK1/2 signaling pathway and EMT-related genes (E-cadherin and N-cadherin) were detected by real-time PCR and Western blot analyses. Results ACT reduced the activity of HCCLM3 cells and inhibited the proliferation of HCC cells, and the effects had certain correlation with drug concentration and time. ACT inhibited the migration and invasion process of HCCLM3 cells in a concentration-dependent manner. ACT downregulated the mRNA and protein expression of genes related to the ERK1/2 signaling pathway. It increased the mRNA and protein expression levels of the EMT-related gene E-cadherin but decreased those of N-cadherin. Conclusion ACT could inhibit EMT and the invasion and migration of HCCLM3 cells in human hepatoma, and the underlying mechanism is closely related to the downregulation of the ERK1/2 signaling pathway.
ABSTRACT
Objective:To explore the anti-tumor molecular mechanism of berberine (BBR)by observing and analyzing its effect on proliferation, apoptosis and autophagy-related gene expression for HCCLM3 cells under high glucose condition. Method:HCCLM3 cells were added into low, medium or high-concentration groups of glucose. It was found in cell counting kit-8(CCK-8)that the high concentration of glucose had the most obvious effect on HCCLM3 cells proliferation. Based on the above experimental result, HCCLM3 cells treated with high concentration of glucose was selected and then different concentrations of berberine (5, 10, 20, 30, 40, 50 μmol·L-1) was added for in vitro intervention for 24 h. Then the effect of each drug group on the proliferation of HCCLM3 cells were studied. At the same time, the control group of metformin was arranged. After that, the changes of apoptosis rate were observed by flow cytometry, and the expression levels of B-cell lymphoma-2(Bcl-2)and autophagy genes Atg5, Beclin1 were detced by Real-time polymerase chain reaction (Real-time PCR). Result:With the increase of glucose concentration, HCCLM3 cell had the strongest migration and proliferation ability in high glucose group (P-1), and the inhibition rate was 33.86%, 40.75% (PPPP PConclusion:BBR could inhibit the proliferation of HCCLM3 cells in high glucose environment. Its inhibition effect for HCCLM3 cells might be achieved by inducing apoptosis of the cells, regulating Bcl-2 and up-regulating the expression levels of autophagy gene Beclin1 and Atg5.Thus BBR plays an anti-tumor role through promoting autophagy in high glucose environment.