马鞭草有效成分对人绒毛膜癌耐药细胞株JAR/VP16的逆转作用研究

Reversal of multidrug resistance in drug-resistance human choriocarcinoma cell line JAR/VP16 by effective component of Verbena Officinalis L

目的:研究马鞭草有效成分4’-甲醚-黄芩素(4’-methylether-scutellarein,4’-M-S)对人绒毛膜癌多药耐药细胞株JAR/VP16的耐药逆转作用。方法:MTT法检测4’-M-S对JAR/VP16细胞的增殖抑制作用,确定药物逆转浓度及其对耐药细胞的逆转倍数和逆转效率;单克隆抗体酶免定量法测定耐药细胞中绒毛膜促性腺激素(hCG)分泌变化;流式细胞术检测细胞周期分布及细胞凋亡率;光镜和透射电镜观察耐药细胞形态学改变。结果:4’-M-S对JAR/VP16细胞具有明显增殖抑制作用,20μg/ml4’-M-S可显著逆转JAR/VP16细胞对鬼臼乙叉甙(VP16)、氨甲喋呤(MTX)及放线菌素D(ACTD)的耐受性,逆转倍数为:5.02、3.67、2.48;逆转效率为:81.19%、76.89%、64.08%;4’-M-S联合作用后,耐药细胞hCG分泌量明显下降;细胞周期阻滞于G0～G1期和G2～M期,细胞凋亡率达19.71%;透射电镜观察发现细胞质空泡化、内质网扩张、核物质浓缩边聚等早期凋亡现象。结论:4’-M-S对JAR/VP16细胞具有耐药逆转作用,可能的机制是抑制耐药细胞生长及诱导细胞凋亡。

Objective：To investigate the reversal effect of the effective component of Verbena Officinalis L. （4＇-methery-scutellarein, 4＇ -M-S） on muhidrug resistant JAR/VP16 cell line. Methods： MTT [ 3- （4,5-dimethylthiazol-2-yl） 2,5-dipheny tetrasodium bromide ] assay was used to examine the action of 4＇-M-S on proliferation of JAR/VP16 cells, determine the available reversal concentration of 4＇-M-S and measure the reversal index and ratio on JAR/VP16 cells. Secretory volume of chorionic gonadotrophic hormone （hCG） was measured by monoclonal antibody enzyme-linked immunoquantitation. Cell cycle profile and apoptotic cell rate was examined by flow cytometry （FCM）, the morphologic and ultramicrostructural changes were observed with light and transmission electronmicroscope （TEM）. Results：The resistance of JAR/VP16 to VP16, MTX and ACTD, was enhanced up to 5.02,3.67,2.48 folds and ratios of 81.19% ,76.89% ,64.08% after exposure of the cells to 20 μg/ml 4＇-M-S, respectively. Secretory volume of hCG in drugresistance cell was decreased obviously when treated with combination of 4＇-M-S and VP16. Cell cycle was blocked at the period of G0-G1 and G2-M , cell apoptotic rate was increased to 19.71%. The earlier apoptotic changes were observed under TEM in the cells treated. Conclusion：4＇-M-S has a strong reversal effect on multidrug resistance in JAR/VP16 cells, which may be achieved by inhibiting growth of JAR/VP16 cells and inducing cell apoptosis.