ABSTRACT
Objective::To investigate the effect of serum containing Yanghetang (YHT) on the apoptosis of MCF-7 cells in breast cancer based on the mitogen-activated protein kinase (p38)/signal transduction and transcriptional activator 3 (STAT3) signal pathway. Method::YHT liquid with crude drug 1 g·mL-1 was prepared. Female SD rats were randomly divided into control group (distilled water), and high, medium and low-dose YHT groups (24, 12, 6 g·kg-1). YHT-medicated serum was prepared, and 10%medicated serum was used to intervene MCF-7 cells. Cell proliferation and cytotoxicity assay (CCK-8) was used to detect the effect of serum containing YHT on MCF-7 cell proliferation, apoptosis of MCF-7 cells was detected by flow cytometry protein expressions of p38 and STAT3 were detected by Western blot, Quantitative Real-time PCR (Real-time PCR) was used to detect the expressions of B-cell lymphoma/leukemia-xl(Bcl-xl) and Survivin mRNA. Result::CCK-8 assay showed that YHT serum inhibited the proliferation of MCF-7 cells in a time and dose-dependent manner compared with the blank group. The inhibitory effect was most obvious in the high-dose group, with the inhibition rates of 38%, 45%and 54%at different time points (P<0.01). Flow cytometry showed that, compared with the blank group, the apoptosis rate in the medium and high-dose groups increased significantly in a dose-dependent manner, with the apoptosis rates at 11.6%and 16.5%respectively (P<0.05, P<0.01). Western blot analysis showed that, compared with the blank group, the expressions of p38 and STAT3 protein was decreased in high, medium-dose YHT groups (P<0.01) in a dose-dependent manner. Compared with the blank group, the expressions of Bcl-xl and Survivin mRNA were decreased in high, medium-dose YHT groups (P<0.05, P<0.01) in a dose-dependent manner. Conclusion::YHT serum can promote the apoptosis of MCF-7 cells in breast cancer, which may be related to the p38/ STAT3 signaling pathway.
ABSTRACT
OBJECTIVE:To study the effect and its mechanism of carvedilol on leptin-induced activation and proliferation of LX2 human hepatic stellate cells(HSC-LX2). METHODS:HSC-LX2 with logarithmic growth periods were divided into blank con-trol group,leptin-stimulated group and carvedilol low-concentration,medium-concentration,high-concentration groups(5,10,20μmol/L). Except for the blank control group,other groups were added 0.1 g/L leptin and corresponding concentration of carvedilol. After 24 h,MTT method was used to detect the optical density(OD)value of cells and calculate the proliferation rate. Flow cytom-etry was used to detect the cell cycle and apoptosis. Real-time fluorescence quantitative polymerase chain reaction method was used to detect the α-smooth muscle actin (α-SMA),matrix metalloproteinase inhibition factor 1 (TIMP-1),leptin,leptin receptor mRNA expressions. Western blot method was used to detect phosphorylated Janus kinase 2(p-JAK2),phosphorylated signal trans-duction and transcriptional activator 3 (p-STAT3) protein expressions. RESULTS:Compared with blank control group,OD value of cell was increased in leptin-stimulated group;apoptotic rate was decreased;cells of G0/G1 were decreased;α-SMA,TIMP-1, leptin,leptin receptor mRNA expressions and p-JAK2,p-STAT3 protein expressions were increased (P<0.05). Compared with leptin-stimulated group,OD values of cells were decreased in carvedilol concentration groups;apoptotic rate was increased,and the cells were mainly blocked in G0/G1 phase;α-SMA,TIMP-1,leptin,leptin receptor mRNA expressions and p-JAK2,p-STAT3 protein expressions were decreased(P<0.05)and was concentration-depended(P<0.05). CONCLUSIONS:Carvedilol can inhibit the activation and proliferation of leptin-induced HSC-LX2,promote its apoptosis. The mechanism may associate with down-regulat-ing leptin,leptin receptor gene expression and blocking JAK2/STAT3 signal pathway activation by leptin in cells.