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目的:探讨华蟾素(cinobufacini)对人乳腺癌细胞株MDA-MB-231增殖、细胞周期和体外侵袭的影响及其可能机制。方法:用CCK-8试剂盒测定并绘制华蟾素作用后MDA-MB-231细胞的生长曲线,FCM法分析细胞周期分布,Transwell小室检测MDA-MB-231细胞的体外侵袭能力,RT-PCR检测细胞周期素(cyclin)及p21mRNA的表达变化。结果:华蟾素可抑制MDA-MB-231细胞的增殖能力,其半数抑制浓度(half inhibition concentration,IC50)为0.31mg/mL,抑制作用随着华蟾素作用时间的延长而增强,与对照组相比差异有统计学意义(P<0.05);Transwell小室检测表明,华蟾素作用后细胞的侵袭能力比对照组明显下降(P<0.05);FCM分析可见,随着华蟾素浓度的增加,停滞于S期的细胞比率比对照组明显增加(P<0.0001);华蟾素作用后,MDA-MB-231细胞cyclin A1、cyclin D1和cyclin E1 mRNA的表达水平下降,p21 mRNA的表达水平上升,但cyclin B1 mRNA的表达无明显改变。结论:华蟾素通过调控cyclin A1、cyclin D1、cyclin E1和p21的表达而抑制乳腺癌细胞的增殖和侵袭,并影响其细胞周期的分布。
Objective: To investigate the effects of cinobufacini on the proliferation, cell cycle and in vitro invasion of human breast cancer cell line MDA-MB-231 and its possible mechanism. Methods: The growth curve of MDA-MB-231 cells was measured and plotted with CCK-8 kit. The cell cycle distribution was analyzed by FCM. The invasion ability of MDA-MB-231 cells in vitro was detected by Transwell chamber. The changes of cyclin and p21 mRNA expression were examined. Results: Cinobufacini inhibited the proliferation of MDA-MB-231 cells with an IC50 of 0.31 mg / mL. The inhibitory effect increased with the increase of cinobufacini injection time (P <0.05). Transwell chamber assay showed that the invasion ability of cells after cinobufotalin treatment was significantly lower than that of the control group (P <0.05). FCM analysis showed that with the increase of cinobufacini concentration (P <0.0001). After cinobufagin treatment, the expression of cyclin A1, cyclin D1 and cyclin E1 mRNA in MDA-MB-231 cells decreased and the expression of p21 mRNA However, the expression of cyclin B1 mRNA did not change significantly. CONCLUSION: Cinobufacine inhibits the proliferation and invasion of breast cancer cells by regulating the expression of cyclin A1, cyclin D1, cyclin E1 and p21, and affects their cell cycle distribution.