紫草素对人绒癌耐药细胞株增殖的抑制及凋亡的作用

Regulatory effects of shikonin on inhibiting proliferation and inducing apoptosis of drug-resistant choriocarcinoma cell line JAR/MTX

目的研究紫草素对体外培养的人绒癌耐药细胞株(JAR/MTX)抑制增殖、促进凋亡及诱导细胞周期停滞的作用。方法经不同浓度的紫草素分别作用24 h、48 h、72 h后,应用cck-8试剂盒检测其细胞毒性,应用流式细胞仪分析细胞周期分布及凋亡率变化,应用荧光显微镜观察凋亡现象。结果当紫草素浓度从0.1875μg/ml增加到3μg/ml时,其48 h抑制率从46.48±12.0%升高到92.05±9.2%,0.75μg/ml紫草素分别作用24 h、48 h、72 h,其抑制率分别为43.94±10.1%、68.56±10.2%、84.97±18.63%,与对照组相比差异具有显著性(P<0.05或P<0.01),可见随着紫草素剂量的增加和作用时间的延长,对绒癌耐药细胞生长的抑制率也明显增加。当紫草素浓度从0.1875μg/ml增加到3μg/ml时,与细胞作用48 h后,凋亡率由12.13±5.67%升高到30.08±7.85%(P<0.05或P<0.01),随着药物浓度增加凋亡率亦增加,并可见典型的凋亡细胞核形态学变化。细胞周期分布也呈浓度依赖性改变,一方面增高G0/G1期细胞比例(P<0.05),另一方面降低S期和G2/M期细胞比例(P<0.05)。结论紫草素对JAR/MTX具有明确的抗增殖、促凋亡、阻滞细胞周期进程的作用,值得进一步深入研究与探讨。

Objective To determine the effect of shikonin on proliferation and apoptosis of drug-resistant choriocarcinoma cell hne JAR/MTX in vitro.Methods JAR/MTX cells were treated with different concentrations of shikonin for 24 h,48 h and 72 h, respectively. The effect of shikonin on proliferation was detected by cycotoxicity test. The change of apoptosis rate and cell cycle disposition were analyzed by flow cyctometry assay, Cell apoptotic phenomenon was observed by fluorescence test. Results Cultared with the concentration of shikonin from 0. 1875 μg/ml to 3 pg/ml for 48 h, the inhibition rate increased from 46.48 ± 12.0% to92.05 ± 9.2 %. Cultured with the concentration of 0.75 μg/ml for 24 h, 48 h, 72 h, the inhibition rate were 43.94 ± 10.1%, 68.56 ± 10.2%,84.97± 18.63% respectively. The difference was statistically significant, with control group（P 〈 0.05 or P 〈 0.01）. Shikonin inhibited the proliferation of JAR/MTX cell hne. The effect increased as the increase in concentration of the drug and the duration. The same was to the apoptosis rate which varied from 12.13 ± 5.67 % to 30.08 ±7.85 % （ P 〈 0.05 or P 〈 0.01 ） when the concentration of shikonin from 0.1875 μg/ml to 3 μg/ml for 48 h. Typical apoptotic nuclear morphological changes were observed. Shikonin significantly blocked the cell cycle progression in JAR/MTX cell hne,decreased S,G2/M phase （ P 〈 0.05）and increased G0/G1 phase （ P 〈 0.05）. Conclusion The conclusion is clear that the exact effects of shikonin on inhibiting proliferation,inducing apoptosis and blocking cell cycle. Shikonin is worthy to be further studied.