Journal: International Journal of Molecular Sciences

Article Title: Flavonoids from Orostachys japonicus A. Berger Inhibit the Invasion of LnCaP Prostate Carcinoma Cells by Inactivating Akt and Modulating Tight Junctions

doi: 10.3390/ijms140918407

Figure Lengend Snippet: Tightening tight junctions (TJs) by flavonoids extracted from Orostachys japonicus (FEOJ) through suppression of the expressions of claudin-1 and claudin-3 at the transcriptional level in LnCaP human prostate cancer cells. Du145, LnCaP, and PC3 human prostate carcinoma cells were treated with FEOJ at the indicated concentrations for 48 h. ( A ) Cell viability was estimated by MTT assay; ( B ) TER values were measured. Each point represents the mean ± SD of three independent experiments. Significance was determined by the Student’s t -test ( * p < 0.05 vs. control); ( C ) Total RNA was isolated and reverse-transcribed. Resulting cDNAs were then subjected to PCR. The reaction products were subjected to electrophoresis in a 1% agarose gel and visualized by EtBr staining. GAPDH was used as an internal control; and ( D ) Western blotting was performed using the indicated antibodies and an ECL detection system. Actin was used as an internal control. Data were representative of two independent experiments.

Article Snippet: Du145, LnCaP, and PC3 human prostate carcinoma cells were obtained from American Type Culture Collection (Rockville, MD, USA) and grown in RPMI 1640 supplemented with 10% fetal bovine serum (FBS), 2 mM L-glutamine, 25 mM N -2-hydroxyethylpiperazine- N′ -2-ethanesulfonic acid, 25 mM NaHCO 3 , 100 IU/mL penicillin and 10 μg/ml streptomycin at 37 °C in a humidified atmosphere of 95% air and 5% CO 2 .

Techniques: MTT Assay, Control, Isolation, Reverse Transcription, Electrophoresis, Agarose Gel Electrophoresis, Staining, Western Blot