表没食子儿茶素没食子酸酯对人膀胱癌T24细胞增殖的影响

Effects of EGCG on proliferation of human bladder carcinoma T24 cells

目的研究表没食子儿茶素没食子酸酯((-)-epigallocatechin gallate,EGCG)在体外对人膀胱癌细胞系T24细胞增殖和细胞周期的影响及其可能机制。方法不同浓度的EGCG作用T24细胞24h后,应用MTT法、集落形成试验和碘化丙锭(PI)染色分别观察T24细胞在增殖、集落形成和细胞周期等方面的变化;并检测CyclinD1和CDK4的变化。结果20-100μg/mL EGCG对T24细胞作用24h后的抑制率分别为4%-61%,与对照组相比,当EGCG为40-100μg/mL时抑制率有显著性差异(P<0.05)。EGCG能抑制T24细胞集落形成,当浓度为20和40μg/mL时能完全抑制其细胞集落形成。EGCG能使T24细胞出现G0/G1期阻滞,与对照组相比,在40-80μg/mL时有统计学意义(P<0.05)。EGCG能抑制细胞CyclinD1和CDK4的表达。结论EGCG能显著抑制T24细胞增殖并诱导G0/G1期阻滞,后者可能与抑制CyclinD1和CDK4表达有关。

Abstract： Objective The present study was undertaken to study the effects of （ - ）-epigallocatechin gallate （EGCG） on growth inhibition and cell cycle, and to investigate the corresponding molecular mechanism of T24 cells. Methods The eytotoxie, antiproliferative and cell cycle arrest effects of EGCG on T24 cells were determined with varying concentration of EGCG treatment for 24 hours by MTT assay, anehorage-dependent eolony-forming assay and flow cytometry. The cell cycle modulating protein Cyelin DI and CDK4 were determined by immunoblotting. Results Inhibitory rates of cell viability treated with EGCG at concentrations of 20-100 μg/mL after 24 hours ranged from 4% to 61%. EGCG at dosages of 40, 80and 100 μg/mL inhibited the proliferation of cells significantly（P 〈 0.05 ）. There was a drastic decrease in the ability of the T24 calls to form colonies with increasing doses of EGCG （ 10 - 40 μg/mL） and EGCG at dosages of 20 and 40 μg/mL completely inhibited the proliferation of cells without any colony forming. The T24 cells treated with EGCG resulted in significant G0/Gl-phase cell cycle arrest and dose-dependent downmodulation of the protein expression of Cyclin D1 and CDK4. Conclusion EGCG could significantly inhibit proliferation and induce G0/G1 -phase cell cycle arrest of T24 cells which might be relative to prohibition of Cyclin D1 and CDK4 expression.