Fig 1.
Tetrandrine (Tet) inhibits human RCC proliferation A. The chemical structure of Tet. The OD value and inhibition rate of Tet on viability of RCC 786-O (B, C) and 769-P (D, E) cells. Cells under 90% cell density were treated with the vehicle control or Tet at the indicated concentrations for 24h. The dynamic OD value was assayed at the concentration of 0.5 μM (F). All the cell viability was detected by modified MTT assay. The values were showed as mean±S.E. The experiment was performed in triplicate.
Fig 2.
Tet suppresses cell migration and invasion of human RCC.
The inhibition of Tet on 786-O (A) and 769-P (B) cells were determined by wound closure assay. The width of scratch was measured in vehicle or Tet group. 786-O (C) and 769-P (D) cells were treated with vehicle or Tet for 24 h using the Transwell migration and invasion assay. The quantitative data were shown in the right panel. The values were showed as mean ± S.E. All the experiments were performed in triplicate (** means P<0.05, *** means P<0.01).
Fig 3.
Tet markedly decreases phospho-AKT, NF-κB and MMP-9 expression in human RCC.
786-O (A) and 769-P (B) cells treated with vehicle or Tet (0.1μM to 0.5μM) for 24 h were immunoblotted for the expression of AKT, phospho-AKT, NF-κB and MMP-9. β-actin was used as a loading control. The quantitative data were shown in the lower panel (C,D). The values were showed as mean±S.E. Representative results from three independent experiments were shown (** means P<0.05, *** means P<0.01).
Fig 4.
Tet inhibits RCC migration and invasion via decreasing Akt, NF-κB and MMP-9 expression.
The metastatic effects of 786-O (A) and 769-P (B) were detected by transwell assay under Tet treatment (0.5μM), with or without LY294002 (LY, 20μM) for 24 h. The protein levels of Akt, phospho-Akt, NF-κB and MMP-9 in 786-O (C) and 769-P (D) cells were detected after indicated treatments by western blotting. And the experiments were performed in triplicate.
Fig 5.
Tet represses cell migration and invasion of human RCC by negatively regulating NF-κB expression.
The metastatic phenotype of 786-O and 769-P was determined by transwell assay. The migrated and invaded 786-O (A) and 769-P (B) cells were counted after treatment with Tet (0.5μM), PDTC (10μM), or the both for 24 h. Under similar treatment, lysates from 786-O (C) and 769-P (D) cells treated with Tet or PDTC were immunoblotted for MMP-9 and NF-κB. All the experiments were performed in triplicate. Representative results from three independent experiments were shown.