Plumbagin Inhibits Proliferation, Invasion And Migration Of Bladder Cancer Cells By Decreasing Productions Of Twist And Matrix Metalloproteinase-9

Objective:To explore the growth inhibition and apoptosis inducing effects of Plumbagin on human bladder cancer T-24 cell line. Methods:After administration of 1-20 umol/L Plumbagin for 24-72 h, cell proliferation of T-24 cells were studied by MTT colorimetry. Cell proliferation ability also was detected by colony formation assay. Cellular apoptosis was inspected by acridine orange-ethidium bromide fluorescent staining, Hoechst 33258 fluorescent staining, and Annexin V-FITC and propidium iodide staining flow cytometry(FCM). Results:Plumbagin could inhibit the growth of T-24 in a dose-and time-dependent manner. After administration of 1 to 20 umol/L of Plumbagin for 24 h, the cell proliferation rates of cells decreased 1.7% to 88.1%. After being treated by 1-3 umol/L of Plumbagin, cell clone formation rates were dramatic decline. Partial cells presented the characteristic morphological changes of apoptosis. The cellular apoptotic rates induced by 2.5 umol/L to 10 umol/L Plumbagin for 24 h were 11.2% to 46.2%. Conclusion: Plumbagin could significantly inhibit the growth of T-24 cells by suppressing proliferation and inducing cell apoptosis, which lays the groundwork for further investigation into the mechanisms of anti-tumor effects of Plumbagin. Objective To investigate the inhibitory effects of plumbagin on the invasion and migration and its correlation with Twist and matrix metalloproteinase-9 (MMP-9) in bladder cancer T24 cells. Methods:The cell migration and invasion were measured by Wound healing assay and Transwell chamber assay. The mRNA and protein levels of nuclear factorκB (NF-κB), Twsit and MMP-9 were evaluated by semiquantitative reverse transcription polymerase chain reaction and western blotting. Also, NF-κB nuclear translocation were analyzed by NF-κB Activation, Nuclear Translocation Assay Kit. Results:In this study, 1-5uM plumbagin had exhibited an inhibitory effect on the abilities of migration and invasion (P< 0.05). The mRNA and protein levels of NF-κB, Twsit and MMP-9 were significantly reduced by 1-10uM plumbagin treatment. Next, lOuM plumbagin significantly decreased the nuclear levels of NF-κB. Conclusion:We demonstrated the inhibitory effects of plumbagin on the invasion, migration and of T24 cells, while plumbagin treatment may decrease the expressions NF-κB, Twsit and MMP-9. Objective:Ethanol treatment induces an increase in oxidative stress. As licorice compounds are potent antioxidants, our aim was to examine whether magnesium isoglycyrrhizinate attenuated lipid peroxidation, the major end-point of oxidative damage resulting from ethanol administration. Methods:Four groups(18 animals in each group) of male Kunming mice were used. The first group served as control and received 0.4ml normal saline daily for 18 days orally. The second group of mice was given 56% ethanol at 16ml/kg body weight per day for 18 days orally. The third group was given the same dose of ethanol and administrated magnesium isoglycyrrhizinate (15mg/kg.d, i.p.) for 18 days. The fourth group was given the same dose of ethanol and administrated with magnesium isoglycyrrhizinate (45m g/kg.d, i.p.) for 18 days. Twenty four hours after 9 days or 18 days of treatment the mice were sacrificed using 10% chloral hydrate. Sperm counts and motility in the epididymis were assessed. The lipid peroxidation and antioxidants of testicular mitochondria were also determined. The pathological changes of testicle tissue of the mice were observed by light microscopy. Results:Magnesium isoglycyrrhizinate effectively prevented the ethanol-induced seminiferous epithelium disorganization and degeneration of Sertoli cells and germ cells. Sperm counts and motility of the magnesium isoglycyrrhizinate treated groups were higher than those of the alcohol treated group, but were lower than those of the control group. The drug exhibited an ability to counteract ethanol induced oxidative challenge as it effectively reduced testicular malondialdehyde (MDA) and increased the activities of superoxide dismutase and glutathione peroxidase Conclusion: Magnesium isoglycyrrhizinate is able to inhibit the ethanol-induced lipid peroxidation and has a protective effect against testicular oxidative injury. Objective:To study the antioxidant effects of the Magnesium isoglycyrrhizinate on hydrogen peroxide-mediated sperm oxidative damage in vitro. Methods:Pre-treatment fertile male semen suspensions 30min with Magnesium isoglycyrrhizinate at doses of 20ug/ml-2.5mg/ml and 600 um ascorbic acid,200uM hydrogen peroxide induced oxidative damage, then assessed sperm viability, apoptosis rate and sperm DNA integrity after 90min by eosin-Y, Wright-Giemsa staining and acridine orange fluorescence staining. Results: 20ug/ml-0.5mg/ml significantly protected sperm DNA against oxidative damage,600 um ascorbic acid also protected(P<0.05).2.5 mg/ml Magnesium isoglycyrrhizinate had no obvious protective effect(P>0.05). Compared with ascorbic acid added at physiological concentrations(600uM),0.1mg/ml Magnesium isoglycyrrhizinate was the most potent antioxidant, followed by 0.5 mg/ml Magnesium isoglycyrrhizinate, ascorbic acid,20ug/ml Magnesium isoglycyrrhizinate. Conclusion:Magnesium isoglycyrrhizinate reduces hydrogen peroxide-mediated sperm oxidative damage in vitro. Magnesium isoglycyrrhizinate may have a role to play in antioxidant protection against male infertility.