人骨肉瘤细胞体外培养与桂皮酸诱导分化的实验研究

Induced-differentiation effects of cinnamic acid on human osteogenic sarcoma cells cultured primarily in vitro

目的:体外培养手术切取人骨肉瘤标本并由植物成分桂皮酸(cinnamic acid,CINN)处理,研究其对肿瘤细胞增殖的抑制作用及诱导分化作用。方法:应用原代法培养手术切除标本,传代至第10代获得相当的细胞量后,设立对照组使用CINN处理,对各组骨肉瘤细胞进行形态学观察,组织化学染色,免疫组织化学染色,流式细胞仪(FCM)检测细胞周期和裸鼠移植瘤试验。结果:以浓度为2×10-3mol/L的CINN处理人骨肉瘤细胞,其存活细胞形态学观察和染色结果均显示分化改变,增殖抑制率第2天为23.3%,第4天为41.7%,第6天为50.6%。FCM检测显示,对照组细胞G1期72.5%,G2期24.9%,S期2.6%;实验组细胞G1期60.9%,G2期9.6%,S期29.5%。处理后的细胞裸鼠移植瘤生长较对照组缓慢,切片呈现大片坏死。结论:原代法体外培养人骨肉瘤细胞成功,体外实验CINN有抑制骨肉瘤细胞增殖和诱导其分化的作用。

OBJECTIVE： To culture specimens of human osteogenic sarcoma primarily in vitro, and study the inhibition and induced-differentiation of survived human osteogenic sarcoma cells induced by cinnamic acid, one of the plant components. METHODS： Human osteogenic sarcoma cells were cultured and treated by different doses of drugs in vitro and their induced-differentiation effects were observed in terms of morphology, histochemistry and immunohistochemistry. Cell cycle changes were observed by Flow cytometry （FCM）. Transplantation essay in nude mouse was also performed. RESULTS：The experiment concentration of CINN was 2 × 10^-3 mol/L. The proliferation inhibition rates are 23. 3%, 41.7% and 50.6% respectively in 2,4 and 6 days after administration. FCM demonstrated that the percentage of cell cycle was G1 phase 72. 5%, G2 phase 24. 9%, S phase 2. 6% in the control group while in the test group the percentage of cell cycle was G1 phase 60.9%, G2 phase 9.6%, S phase 29.5%. The drug-treated cells transplantation in nude mouse grew slower than the control group. CONCLUSION：Human osteogenie sarcoma cells are cultred successfully in vitro and CINN has the inhibitory and induced-differentiation on human osteogenic sarcoma cells.