ABSTRACT
Objective:To investigate the mechanism in growth inhibition of garcinia acid to kidney cancer cell lines ( RC-2 ) . Methods:RC-2 cells were cultured in vitro, and garcinia acid at various concentrations was co-cultured with RC-2 cells. The antipro-liferative activities were determined by CCK-8 assay, the percentage of apoptosis was determined by flow cytometry analysis, and the expression levels of Survivin and related proteins in Wnt3α/β-catenin signal pathway were measured using Western Blot analysis. Re-sults:After the treatment with garcinia acid for 24h and 48h, the proliferation of RC-2 cells was significantly suppressed by garcinia acid in a dose-dependent manner, and all garcinia acid groups had significantly higher apoptosis percentage of RC-2 cells than the con-trol group. In G0/G1 period, RC-2 cells reduced while increased in G2/M period, and in S period, the cells remained the same a-mount. The expression levels of Survivin, Wnt3α and β-catenin decreased in RC-2 cells after the treatment with garcinia acid. Con-clusion:Garcinia acid can inhibit the proliferation and promote the apoptosis of RC-2 cells, and the mechanism may be related with the inhibition of Wnt3α/β-catenin signal pathway and further decreasing the expression levels of Survivin.
ABSTRACT
Objective To study the effect and mechanism of increase in radio sensitivity of kidney cancer cells(GRC-1) induced by ?-elemenen in vitro. Methods GRC-1 cells were divided into 3 groups, blank group (added with 2 mL culture medium), emulsion group (added with 2 mL blank emulsion culture medium) and drug group (added with 2 mL 50 mg?L -1 ?-elemenen culture medium). After been cultivated for 24 hours, the cells were irradiated using 6MeV X-linear accelerator in different doses at the rate of 400cGy per minute. Number of cell clones was counted, and radiation-survival curve of GRC-1 cells was drawn. Flow cytometry (FCM) was used to measure cell cycle and apoptosis. Cells of climbing flake were dyed by immunocytochemical method, the gene expression of bcl-2 and PCNA was measured by imaging system. Results The cell cycle showed that the G 2M blocking caused by 50 mg?L -1 ?-elemenen was enhanced with time increase. It reached peak at 24 hours. FCM showed that the level of apoptosis increased with increase in drug dose and action time. The gene expression of bcl-2 was decreased by 20% in drug group than that in blank group, but there was no expression of PCNA in the two groups. Conclusion The radiosensitivity of GRC-1 cells can be enhanced by ?-elemenen. The mechanism of effect may be associated with the cell cycle blocking, inducing cell apoptosis and down-regulating expression of bcl-2 gene.