ABSTRACT
OBJECTIVE：To study the effects of loganin on the prolife ration and apoptosis of liver cancer HepG 2 cells，and to explore its mechanism. METHODS ：CCK-8 assay was used to detect the effects of different concentrations （10，25，50，100， 150，200，300，400 µg/mL）of loganin on the proliferation activity of HepG 2 cells for 24 and 48 h. HepG 2 cells were divided into control group ，loganin low-concentration ，medium-concentration and high-concentration groups （50，100，150 μ g/mL）. After treated for 24 h，morphological changes of apoptosis of cells were detected by Hoechst 33342 fluorescence staining. The apoptosis and cycle distribution of cells were detected by flow cytometry. Western blotting was used to detect protein expression of Cyclin D1， PCNA， Bcl-2， Caspase-3， Cleaved-Caspase-3， Caspase-9 and Cleaved-Caspase- 9. RESULTS ： Loganin inhibited the proliferation of HepG 2 cells，in concentration-dependent trend. Compared with control group ，apoptosis as pyknosis and fragmentation occurred ，and the apoptosis rate increased significantly in loganin low-concentration ，medium-concentration and high-concentration groups （P＜0.01）；the cell were mainly blocked in S phase ；relative protein expression of Cyclin D 1，PCNA and Caspase- 3 were significantly decreased ，while that of Cleaved-Caspase- 3 were significantly increased in loganin low- concentration， medium-concentration and high-concentration groups （P＜0.05 or P＜0.01）； relative protein expression of Cleaved-Caspase-9 were increased significantly ，while that of Bcl- 2 and Caspase- 9 were decreased significantly in loganin medium-concentration and high-concentration groups （P＜0.05 or P＜0.01）. CONCLUSIONS ：Loganin can significantly inhibit the proliferation and induce apoptosis of HepG 2 cells，the mechanism of which may be associated with inhibiting Bcl- 2 protein expression and promoting Caspase- 3，Caspase-9 activation.
ABSTRACT
Objective:To study the anti-proliferative effect of lupeol on human bladder cancer T24 cell line and the regulating mechanism for p53/miR-34a signaling. Methods:CCK-8 assay was performed to evaluate the effects of lupeol at different concentra-tions on cell viability in 24 h and 48 h. Caspase inhibitors were used to identify the subtypes of Caspase during lupeol induced cell death. The effects of lupeol on the expression of total p53 protein and miR-34a were evaluated by western blot and real-time PCR, re-spectively. The effects of lupeol on downstream targets of miR-34a were quantified by real-time PCR. Results:Lupeol could inhibit the proliferation of T24 cells in a dose-dependent manner. The IC50 of lupeol was (77. 23 ± 6. 78) μmol·L-1 in 24 h. Compared with the control group, lupeol could elevate the expressions of p53 and miR-34a (P<0. 01). Moreover, the mRNA expression of miR-34a tar-gets, Bcl-2, CD44 and c-Myc were significantly down-regulated after the treatment with lupeol (P<0. 01). Conclusion:Lupeol can inhibit T24 cell proliferation, which is related with the regulating effects on p53/miR-34a signaling.
ABSTRACT
Objective:To study the mechanism of anti-proliferative effect of lupeol on highly metastatic human hepatocellular car-cinoma HCCLM3 cells. Methods:CCK-8 assay was performed to evaluate the effects of lupeol at different concentration on cell viability in 12-48 h. Caspase inhibitors were used to identify the subtypes of caspases activated during lupeol-induced cell death. The effects of lupeol on the mRNA expression of caspase family and Bcl-2 related genes were detected by real-time PCR. The effects of lupeol on HC-CLM3 cell phase distribution were investigated by flow cytometry. Results:Compared with the control group, lupeol could inhibit HC-CLM3 cell proliferation in a concentration-dependent manner with IC50 of 93 μmol·L-1 in 24h. The number of HCCLM3 cells in the period of G2/M was increased by 1-fold when the lupeol concentration was within 60-100 μmol·L-1 . Lupeol could activate the path-way of caspase, and the mRNA expression of caspase-3 was elevated by 50%-150% when compared with that in the control group. Mo-reover, the mRNA expression of p53 and Bax were increased above 1-fold by lupeol at 100 μmol·L-1 , and the Bcl-2 and PARP ex-pression were significantly suppressed by lupeol at 60-100 μmol·L-1(P<0. 05 or P<0. 01). Conclusion:The results indicate that lupeol has anti-proliferative effect on the liver cancer cells, which is beneficial to the prevention and treatment of liver cancer.