染料木黄酮对骨肉瘤细胞增殖及侵袭的影响及作用机制

Effects of Genistein on proliferation and invasion of osteosarcoma cells and action mechanism

目的探讨染料木黄酮对人骨肉瘤MG-63细胞增殖、侵袭的影响及其作用机制。方法用染料木黄酮处理MG-63细胞，CCK－8法检测细胞增殖；Transwell小室体外侵袭实验法检测MG－63细胞侵袭能力；实时定量聚合酶链反应（PCR）和蛋白印迹法（Westernblot）分别检测MG－63细胞基质金属蛋白酶（MMP）-2／MMP．9的mRNA及蛋白水平。结果CCK－8法表明染料木黄酮处理组MG-63细胞增殖被明显抑制，抑制率最高（85．3±5．1）％（P〈0．05）；染料木黄酮在0、12．5、25．0、50．0μmol／L时穿膜细胞数分别为（96．5±5．5）、（88．4±3．2）、（53．8±4．4）、（36．6±3．8）个，可明显降低MG－63细胞的侵袭能力（P〈0．05）；染料木黄酮处理后MMP－2及MMP-9的mRNA表达水平明显下降（P〈0．05）；MMP－2／MMP-9蛋白表达水平也明显下降，且具有时间依赖陛。结论染料木黄酮抑制骨肉瘤MG-63细胞增殖及侵袭转移能力，这种作用与MMP－2及MMP－9表达变化有关。

Objective To evaluate the effect of Genistein on the proliferation and invasion of os- teosarcoma cells. Methods CCK-8 was used to detect the proliferation of osteosarcoma cells treated with different concentrations of Genistein. Transwell invasion assay was used to measure the invasion of osteosarcoma cells. The matrix metalloproteinase （MMP） -2/9 expression was analyzed by Western blotting and real-time quantitative-polymerase chain reaction （PCR） in the MG-63 osteosarcoma cells treated with Genistein. Results CCK-8 showed that Genistein could inhibit the proliferation of osteosarcoma cells in a time- and dose-independent manner with the highest inhibition ratio being （85.3 ± 5.1 ） % （P 〈 0.05 ）. Transwell invasion assays revealed that the number of cells was （96. 5 ±5.5 ） , （88.4 ±3.2 ） , （53.8 ±4. 4）, （36. 6 ±3. 8） at the concentrations of 0, 12. 5, 25.0, 50.0 μmol/L Genistein respectively, suggesting Genistein could significantly inhibit the invasion of osteosareoma cells （ P 〈 0. 05 ）. MMP-2/9 rnRNA and protein expression was significantly inhibited by Genistein （ P 〈 0. 05 ） in a time-independent manner. Conclusion Genistein can inhibit proliferation and invasion of osteosarcoma cells in vitro , which is associated with the changes of MMP-2/9 expression.