The Study Of Anti-breast Cancer Mechanism Of Luteolin Through EGFR Signaling Pathway And The Relationship With ERα/IGF-1R

Breast cancer is a kind of multiple malignant tumors,has become one of the fastest rise in malignant tumor.Breast cancer was related with the exception activated of intracellular signaling pathways;The epidermal growth factor receptor(EGFR)is an important tumor protein,can promote cell growth and proliferation.Its mediated signaling pathway play an important role in invasion,and prognosis of breast cancer.Our laboratory earlier research results show that ERa is one of the important targets of luteolin anti-breast cancer.Existing research results also show that the occurrence of breast cancer are closely associated with EGFR signaling pathway,but the relationship between anti-breast cancer effect of luteolin and EGFR pathway has not been reported.So this article mainly system studied the mechanism of luteolin anti-breast cancer effect through EGFR signaling pathways and the relationship with the ER a/IGF-1 R,research results are as follows:1.Through MTT method to detect different concentrations of luteolin on the effect of cell proliferation of MCF-7 cell and MDA-MB-231 cells for 24 h,48 h.The results show that luteolin can significantly inhibit MCF-7 and MDA-MB-231 cells proliferation and in time and dose dependent,compared with control,at 48 h,the inhibition rates of 60 μmol/L luteolin on the proliferation of MCF-7 cells and MDA-MB-231 cells were 44.65 ± 1.36% and33.21±1.25% respectively.The result indicated that the effect on MCF-7 cell is more apparent than MDA-MB-231 cells.2.By MTT method to detect the effect of luteolin on EGF promote MCF-7 cell proliferation.The results showed that compared with EGF treatment alone,after treatment of MCF-7 cells with 60 μmol/L luteolin and 100 μg/L EGF for 24 h,the cell viability was reduced by 55.08± 1.02%.The results indicated that EGF can promote the MCF-7 cell proliferation,and luteolin can significantly inhibit breast cancer cell proliferation induced by EGF.3.By hirst,dyeing and reactive oxygen species testing luteolin on the effect of MCF-7cell apoptosis.The results show that after treated with 60 μmol/L luteolin,cells color more bright,the apoptotic cells increased significantly;the levels of reactive oxygen species increased significantly compared with control group.The results indicated that luteolin can significantly improve the levels of reactive oxygen species in MCF-7 cell,inducing MCF-7cell apoptosis of breast cancer.4.By Western blotting methods detected the effect of luteolin on EGF induced EGFR,AKT,Erk1/2,STAT3 protein phosphorylation.The results show that luteolin can significantly reduce the EGFR,AKT,Erk1/2,STAT3 protein phosphorylation level induced by EGF.Compare with EGF treatment alone,further 40 μmol/L luteolin significantly reduced the phosphorylation of EGFR,AKT,Erk1/2,STAT3 by72.57 ± 1.8% 、 57.02±0.98% 、35.53±0.31%和 45.93±0.62%.The results indicated that Luteolin can inhibit EGFR signaling pathway and its downstream AKT,Erk1/2,STAT3 proteins play an important part role.5.By MTT method to detect the relationship of luteolin inhibit breast cancer cell proliferation induced by EGF with ER a,IGF-1 R signaling.The results show that luteolin and the treatment group with the ER a,IGF-1 R inhibitors can significantly inhibit the proliferation of breast cells induced by EGF,and has no significant difference between them,but have significant influence with ER a and IGF 1 R inhibitor treat alone.Results show that luteolin inhibit breast cancer cell proliferation induced by EGF has no relationship with ER a and IGF-1 R.6.By Western blotting methods detected the relationship between the effect of luteolin on EGFR protein phosphorylation induced by EGF and ER a/IGF-1R.The results show that ER a/IGF-1R inhibitors had no significant effect on EGFR protein phosphorylation level.Luteolin inhibiting EGFR protein phosphorylation level has nothing to do with ER a/IGF-1 R.7.By Western blotting methods detect the relationship between luteolin inhibit ER a/IGF-1R expression and EGFR.The results showed that after joining EGFR blockers,ER a and IGF-1R protein phosphorylation levels compared with EGF group have no significant difference.luteolin inhibit of ER a/IGF-1R protein expression has nothing to do with EGFR8.Through the hemolysis experiment testing luteolin drug toxicity.Results show that the experiment has not been used in various concentration of mignonette cause hemolysis of red blood cells,spectrophotometer results show that the hemolysis rate was < 5%,show reseda element is a safe,non-toxic drug.