摘要
目的:研究山苍子油对体外培养的人乳腺癌细胞MDA-MB-231增殖和凋亡的影响及可能机制。方法:CCK-8比色法和流式细胞术(FCM)分别检测山苍子油对MDA-MB-231细胞增殖和凋亡的影响;Western blot方法检测经山苍子油作用后MDA-MB-231细胞凋亡相关蛋白Bax、Bcl-2、caspase-3、EGFR、p-Akt、Akt蛋白的表达水平;山苍子油、Akt激活剂以及山苍子油联合Akt激活剂作用于MDA-MB-231细胞后Akt、p-Akt蛋白的表达水平。结果:山苍子油对MDA-MB-231细胞的增殖具有明显的抑制作用,与对照组相比具有显著性差异(P<0.05),且其抑制作用呈现浓度和时间依赖性;山苍子油使细胞阻滞于S期并能够诱导细胞凋亡(P<0.01);与对照组比较,随着药物浓度的增加及作用时间的延长,可上调MDA-MB-231细胞促凋亡蛋白Bax、caspase-3的表达,下调抗凋亡蛋白Bcl-2的表达(P<0.01);与对照组比较,随着山苍子油浓度的增加EGFR、Akt、p-Akt蛋白的表达量均随之下降(P<0.01)。Akt激活剂可以明显上调Akt、p-Akt的表达。山苍子油可明显下调被激活的Akt、p-Akt(P<0.01)。结论:山苍子油可显著抑制体外条件下人乳腺癌细胞MDA-MB-231的增殖,呈浓度和时间依赖性;可阻滞MDA-MB-231细胞分裂周期于S期并能诱导细胞凋亡,可能与抑制EGFR/PI3K/Akt信号通路有关。
Objective:To research the effect of Litsea cubeba oil on the human breast cancer cell MDA-MB-231 cultured in vitro and to explore and discuss its possible mechanism and apoptosis of action.Methods:CCK-8 colorimetric method and flow cytometry(FCM)were adopted to detect the effects of Litsea cubeba oil on MDA-MB-231 cell proliferation and on the apoptosis.The expression levels of apoptosis related proteins Bax, Bcl-2,caspase-3,EGFR,p-Akt and Akt after Litsea cubeba oil acted on MDA-MB-231 cells, as well as the expression levels of Akt and p-Akt protein after the Litsea cubeba oil,Akt activator and Litsea cubeba oil combined with Akt activator acted on MDA-MB-231 cells were detected by Western blot method.Results:In vitro experiment, Litsea cubeba oil made a significant inhibitory effect on the proliferation of MDA-MB-231 cells, which was statistically significant(P<0.05)compared with the control group, and the inhibitory effect was concentration-dependent and time-dependent.Litsea cubeba oil blocked the cell cycle in S-Phase and induced apoptosis(P<0.01).Compared with the control group, the expression of pro-apoptotic protein Bax and caspase-3 was regulated up on the MDA-MB-231 cells, and the expression of anti-apoptosis Bcl-2 was regulated down on the MDA-MB-231 cells(P<0.01)with the increase of drug concentration and the extension of action time.Compared with the control group, the expression of EGFR,Akt and p-Akt decreased with the increase of Litsea cubeba oil concentration(P<0.01)with the increase of drug concentration and the extension of action time.The expression of Akt and p-Akt was regulated up significantly by Akt activator.The expression of Akt and p-Ak was regulated down by Litsea cubeba oil(P<0.01).Conclusion:Litsea cubeba oil can significantly inhibit the proliferation of human breast cancer cell MDA-MB-231 cell with concentration dependence and time dependence in vitro.Litsea cubeba oil can block the cell division cycle of MDA-MB-231 cells in S-phase and induce apoptosis.The mechanism of inducing apoptosis may be related to the inhibition of the EGFR/PI3 K/Akt signaling pathway.
作者
周恩正
郭贵明
苏旭
吴春凯
左丽(指导)
ZHOU En-Zheng;GUO Gui-Ming;SU Xu;WU Chun-Kai;ZUO Li(Basic Medical College,Guizhou Medical University,Guiyang 550025,China)
出处
《中国免疫学杂志》
CAS
CSCD
北大核心
2021年第5期571-576,共6页
Chinese Journal of Immunology
