目的探讨积雪草酸对多发性骨髓瘤细胞株RPMI 8226细胞增殖、细胞凋亡、细胞周期的影响及其机制。方法 MTT法检测积雪草酸对肿瘤细胞的抗增殖作用;Hoechest33258染色法检测细胞凋亡的形态学变化;流式细胞术(FCM)检测细胞凋亡及细胞周期的变化;RT-PCR法检测细胞survivin和bcl-2 mRNA表达水平的变化。结果积雪草酸在浓度10~70μmol/L显著抑制RPMI 8226细胞的增殖, 呈时间及浓度依赖性, 作用24、487、2 h的IC₅₀值分别为(42.25±4.57)(、24.88±3.51)(、19.83± 2.88)μmol/L。积雪草酸可诱导细胞发生凋亡, 出现典型的凋亡小体。积雪草酸诱导细胞发生早期凋亡, 并呈浓度依赖性。积雪草酸25~40μmol/L能将RPMI 8226细胞阻滞在G2期, G2期细胞比例随积雪草酸浓度增大而逐渐增高。积雪草酸使survivin和bcl-2 mRNA表达下降, 并与细胞的凋亡程度呈负相关。结论积雪草酸能通过调控细胞周期的进程和诱导细胞凋亡从而抑制RPMI8226细胞增殖, 其诱导凋亡的机制可能与下调survivin和bcl-2的转录水平有关。

Objective To explore the effect of asiatic acid on cell prolifreration,apoptosis,and cells cycle of human multiple myeloma cells RPMI 8226,and its antitumor mechanism.Methods MTT Assay was used to assess the effect of asiatic acid on cells proliferation;Hoechest 33258 staining was studied to evaluate the morphological changes in RPMI 8226 cells induced by asiatic acid;Flow cytometry was used to analyze the induction apoptosis and cell cycle arrest of asiatic acid; RT-PCR was applied to detect the changes of survivin and bcl-2 mRNA expression levels.Results Asiatic acid significantly inhibited the cell proliferation of RPMI 8226 in μmol/L grade level,in a time-and dose-dependent manner.The IC50 values of RPMI 8226 cells were(42.25±4.57),(24.88±3.51) and(19.83±2.88) μmol/L for 24,48,and 72 h.Asiatic acid could induce apoptosis,and cell showed typical apoptotic features.Asiatic acid induced apoptosis in a dose-dependent manner.Cells treated with asiatic acid 25—40 μmol/L were blocked in the G2 phase.The expression of survivin and bcl-2 mRNA was decreased in RPMI 8226 cells,and the extent of apoptosis was a negative correlation with the dose of asiatic acid.Conclusion Asiatic acid could inhibit the proliferation by inducing apoptosis and regulating cell cycle process.The mechanism of induction apoptosis is associated with up-regulation of survivin and bcl-2 transcription level.

国家自然科学基金资助项目(30472267)