The Effect Of NRF2 On The Proliferation Of Breast Cancer Cells By Regulating The Expression Of CXCR4

Objective: To investigate the effect of NRF2 gene targeting and regulation of CXCR4 gene expression in breast cancer cells and its effect on cell proliferation.Method:1.The expression of NRF2 gene and CXCR4 gene in breast cancer cells was changed by plasmid transfection technology.2.The expression level of NRF2 gene mRNA was detected by real-time PCR.3.Western blot was used to detect the protein expression level of CXCR4 gene in breast cancer cells.The effects of up-regulating NRF2 gene,down-regulating NRF2 gene and up-regulating NFR2 gene and down-regulating CXCR4 gene on breast cancer proliferative ability were observed by MTT assay.Group: Group A: MCF-7 cells transfected with NRF2 overexpression plasmid;Group B: MCF-7 cells transfected with shRNA-NRF2;Group C: MCF-7 cells transfected with NRF2 overexpressing + shRNA-CXCR4;Group: negative control group;E group: blank control group.Results: 1.qRT-PCR results: the relative expression level of NRF2 gene in group D was(1.053±0.026),and the relative expression level of NRF2 gene in group E was(1.028±0.060).There was no statistical difference between them.The relative expression level of NRF2 gene was(61.530±0.39),compared with group D,P<0.01,there was significant statistical difference.The relative expression level of NRF2 gene in group B was(0.230±0.03),compared with group D,P<0.01.There was a statistically significant difference;the relative expression level of NRF2 gene in group C was(51.070±1.05),compared with group D,P<0.01,there was a statistically significant difference.2.Western Blot results: The relative expression levels of CXCR4 gene in group D and group E were(0.978±0.078)and(1.033±0.013),respectively,and there was no statistical difference(P= 0.595>0.05).In group A and group D,the relative expression of CXCR4 in group A was(1.829±0.057)higher than that in group D(1.042±0.058)(P=0.0001<0.01).In group B and group D,group B was compared.The relative expression of CXCR4(0.955±0.146)was lower than that of group D(2.153±0.211)(P=0.038<0.05).The relative expression of CXCR4 in group A was compared between group A,group C and group D(2.043).±0.093)was higher than group D(1.435±0.044)(P=0.004<0.01)and group C(0.939±0.071)(P=0.001<0.01).3.Results of MTT method Compared with group E,there was no significant difference in OD between 24 h,48h and 72h(P=0.686(24h),P=0.755(48h),P=0.133(72h)).After 24 h,48h,and 72 h in group A,the OD values were 0.734±0.019,0.961±0.011,and 1.225±0.031,respectively.The difference was significant compared with group D.After 24 h,48h,and 72 h in group B,the OD values were 0.434±0.017,respectively.0.571±0.029,0.702±0.018,the difference was significant compared with the D group;the OD values of the C group were 0.478±0.002,0.641±0.035,and 0.819±0.021 after 24 h,48h,and 72 h,respectively,which was significantly different from the A group.Conclusion: The NRF2 gene may enhance its proliferative capacity by increasing the expression of the CXCR4 gene in breast cancer cells.The NRF2 gene may be a potential target for targeted therapy for breast cancer.