Effect Of Litsea Cubeba Oil On Proliferation And Apoptosis Of Breast Cancer Cell MDA-MB-231 And Its Possible Mechanism

Purpose: To research the effect of Litsea cubeba oil on the proliferation of human breast cancer cell MDA-MB-231 cultured in vitro and on the tumor growth and metastasis in tumor-bearing nude mouse model of breast cancer,and to explore and discuss its possible mechanism 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 and its cycle respectively.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(insulin)and Litsea cubeba oil combined with Akt activator acted on MDA-MB-231 cells were detected by Western blot method.BALB/ C nude mice model of breast cancer was established with human breast cancer cell MDA-MB-231 in vivo experiments.The nude mice were divided into seven groups,including control(normal saline)group,3% DMSO group,cisplatin group,blank group,high concentration group,medium concentration group and low concentration group.Intraperitoneal injection was given for each group.The tumor size,weight change and survival situation of the nude mice were recorded regularly.Tumor inhibition rate and survival rate were calculated.HE staining was used to detect tumor metastasis in liver and lung.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.Akt activator can significantly up regulate the expression of Akt and p-Akt.Litsea cubeba oil can significantly down regulate activated Akt and p-akt(P < 0.01).The average weight of the concentration group of Litsea cubeba oil was the lightest,about 18.34 g,in vivo experiments.Compared with the control group,the inhibition rate of cisplatin group on tumor growth in nude mice was about 73.540% ± 3.38,but there was no inhibition effect of Litsea cubeba oil group(P > 0.05).Compared with the control group,the survival rate of tea polyphenols group was lower.Compared with the control group,there was no significant difference in liver and lung metastasis in the high concentration group,the middle concentration group and the low concentration group(P > 0.05).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/PI3K/Akt signaling pathway.Litsea cubeba oil had no inhibitory effect on the tumor growth and metastasis in nude mice bearing tumor of human breast cancer cell MDA-MB-231.