肉桂酸对人骨肉瘤MG-63细胞增殖和分化的影响

Effect of cinnamic acid on proliferation and differentiation of human osteosarcoma MG-63 cells

目的研究中药有效成分肉桂酸对人骨肉瘤MG-63细胞增殖和分化的影响。方法肉桂酸处理MG-63细胞前后,台盼蓝拒染法进行活细胞计数;流式细胞仪检测细胞周期时相变化;光镜和电镜观察细胞形态和超微结构;VanGieson染色检测细胞Ⅰ型胶原表达;茜素红染色观察钙化骨结节形成;免疫细胞化学检测骨粘素和骨钙蛋白表达。结果肉桂酸明显抑制MG-63细胞增殖,D7抑制率达54.94%±4.69%。周期分析结果显示,肉桂酸处理后G0/G1期细胞比例较对照组明显升高(P<0.01)。光镜和电镜结果显示,肉桂酸处理组MG-63细胞形态和超微结构发生明显改变,呈现相应正常细胞的形态和超微结构特征。肉桂酸促进成骨细胞分化标志物Ⅰ型胶原、骨粘素和骨钙蛋白的表达与钙沉积以及典型骨结节的形成。结论肉桂酸可抑制人骨肉瘤MG-63细胞增殖并诱导其向成骨细胞分化。

Aim To investigate the effect of cinnamic acid on proliferation and differentiation of human osteosarcoma MG-63 cells. Methods MG-63 cells were treated with cinnamic acid. The living cells were counted by trypan blue staining. The morphology and ultrastructure of MG-63 cells were studied with optical microscope and electron microscope. The changes of cell cycle were analyzed by flow cytometry （FCM）. The expression of type I collagen was detected by Van Gieson staining. The formation of mineralized bone nodules was evaluated by alizarin red staining. The expressions of osteocalcin and osteoneetin were detected by immunocytochemical staining. Results Cinnamic acid inhibited the proliferation of MG-63 cells significantly, the inhibitory rate was 54.94% ±4.69% after treatment with cinnamic acid for 7 days. Flow cytometry aalysis showed that cinnamic acid increased the cell numbers in G0/G1 phase significantly（P 〈 0.01 ）. Un- der optical microscope and electron microscope, the morphology and uhrastructure of MG-63 ceils treated with cinnamic acid changed significantly compared with those in control group, showing the characteristics of normal cells. Furthermore, cinnamic acid promoted the expressions of osteoblast markers such as type I collagen, osteonectin and osteocalcin, increased calcium aggradation and facilitated the formation of bone nodules. Conclusion Cinnamic acid can inhibit the proliferation of human osteosarcoma MG-63 cells and induce the differentiation of MG-63 ceils into osteoblasts.