羊齿天门冬根茎提取物对人骨肉瘤细胞增殖的抑制作用

Inhibitory effect of extracts from asparagus filicinus rhizome on proliferation of human osteosarcoma cells

目的探讨羊齿天门冬根茎提取物对人骨肉瘤细胞增殖的影响及其分子机制。方法 MTT法检测羊齿天门冬根茎提取物对人骨肉瘤Saos-2细胞增殖和生长的影响;平皿集落形成法检测羊齿天门冬根茎提取物对细胞锚定依赖性生长的影响;碘化丙啶单染流式细胞仪检测细胞周期分布改变;Western blot检测环氧化酶2(COX-2)蛋白表达水平改变。结果羊齿天门冬根茎乙酸乙酯部位(AF-A)对Saos-2细胞具有细胞毒作用(IC50值为26.7μg/mL);AF-A对Saos-2细胞锚定依赖性生长具有抑制作用,呈剂量相关性(P<0.05);AF-A(30.0、100.0μg/mL)处理Saos-2细胞48h后,细胞S期百分比分别为(43.7±2.5)%和(51.9±1.9)%,与对照组(31.8±4.8)%比较差异有统计学意义(P<0.05);不同浓度AF-A均能抑制Saos-2细胞COX-2蛋白水平表达。结论 AF-A对人骨肉瘤细胞增殖和生长具有抑制作用,其机制可能与诱导人骨肉瘤细胞S期阻滞和抑制COX-2蛋白水平表达有关。

Objective To investigate the inhibitory effect of extracts from asparagus filicinus rhizome on prolifieration of human osteosarcoma Saos-2 cells and its molecular mechanism. Methods MTT assay was used to detect the cytotoxic activity and growth inhibition of three different extracts from asparagus filicinus rhizome against Saos-2 cells; plate colony formation assay was per- formed to detect active fraction of asparagus filicinus rhizome on the anchorage dependent growth of Saos-2 cells ; the cell cycle alter- ation was determined by propidium iodide staining and {low cytometry analysis;the alteration ofprotein expression level of COX-2 was determined by using Western blotting. Results Ethyl acetate fraction of asparagus filicinus rhizome（AF-A） exerted the potent eytotoxicity on Saos-2 cells（ICs0 = 26.7 〉g/mL） ;AF-A induced the inhibitory effect on the anchorage dependent growth of Saos-2 cells in a dose dependent manner（P〈0.05） ;Saos-2 cells treated by AF-A at the concentration of 30.0 and 100.0μg/mL for 48 h induced the increase of percentages of S phage from（31.8± 4.8）% in the control group to（43.7±2.5）%and（51.9 ±1.9）%, the difference showing statistical significance （P〈0. 05）. Western blotting showed that AF-A at different concentrations decreased COX-2 protein expression. Conclusion AF-A posseses the inhibitory effect on the proliferation and growth of human osteosarcoma cells in vitro,and its mechanism might be associated with the induction of S phage arrest and the inhibition of COX-2 protein ex- pression level.