长春花对胆管癌细胞增殖和细胞周期的影响

The effect of extracts from Catharanthus roseus on cholangiocarcinoma in vitro

目的观察长春花草提取物对体外培养的人胆管癌QBC939细胞的抑制作用。方法以长春花全草为原料进行分离提取，获得总生物碱层和水层两种提取物，分别采用噻唑蓝（MTT）法、普通倒置光学显微镜观察法和流式细胞术研究两种提取物对QBC939细胞增殖、细胞形态以及细胞周期的影响。结果长春花两种提取物均能够显著抑制胆管癌细胞的增殖，尤以总碱层的抑制作用最强，半抑制常数IC50分别为0．16mg／L和49．02mg／L；两种提取物均能使细胞的形态发生明显改变。随着作用浓度的增大，长春花碱层作用的细胞G0／G1期从对照组的53．10％减少到5．87％，G2／M期细胞从对照组的26．4％增加到71．8％，Sub-G1期变化不明显；长春花水层提取物作用的细胞G0／G1期也明显减少，G2／M期细胞明显增多，而Sub—G1期则变化明显，出现明显的凋亡峰。结论长春花提取物具有抑制胆管癌细胞增殖、改变细胞形态及细胞周期分布的作用。

Objective To investigate the potential inhibitory effects of herbal Catharanthus roseus on cholangiocarcinoma in vitro. Methods The whole herbal of Catharanthus roseus was used as the material, and total alkaloid extract and water extract were obtained. The effects of the two extracts on human cholangiocarcinoma QBC939 cells were investigated. The inhibition rate of QBC939 cells was measured by methyl thiazolyl tetrazolium （MTr） method. The morphological change of cells was examined by inverted microscopy. The effect on cell cycles was observed by flow cytometry. Results The two extracts could both inhibit the growth of QBC939 cells, but the total alkaloid extract showed stronger proliferation inhibitory effect comparing to the water extract. IC50 value of total alkaloid and water extract was 0. 16 and 49. 02 mg/L, respectively. Obvious morphological changes were both observed after adding the two extracts. Both the two extracts could change the cell cycle of cholangiocarcinoma cells significantly. The ratio of G0/G1 phase cells was decreased greatly from 53.10% to 5.87% and that of G2/M phase cells was increased greatly from 26.4% to 71.8%, respectively. Compared to the ceils treated with the total alkaloid extract, the ratio of Sub-G1 phase cells treated with the water extract changed greatly. The water extract could induce the apoptosis of the cells. Conclusion The extracts from Catharanthus roseus showed the inhibitory effects on proliferation of cholangiocarcinoma cells. They also could induce the morphological change and cell arrest.