Fig. 5

Effects of CYP1A1 overexpression on THC anti-breast cancer. A After synthesis of the overexpressed plasmid CYP1A1, MCF-7 and 4T1 cancer cells were transfected with Lipo2000. Changes of mRNA expression levels of CYP1A1 in MCF-7 and 4T1 were mesasured. B The MCF-7 and 4T1 cancer cells were divided into four groups: Vehicle, THC (80 μM), CYP1A1 and CYP1A1 + THC (80 μM). The expression of CYP1A1, NF-κB associated protein and PD-L1 in each group were detected by RT–PCR and WB experiments. C Immunofluorescence analysis of nuclear transportation of NF-κB protein in MDA-MB-231 cells. D CCk8 assay was used to analyze the effect of transfection of overexpressed plasmid on the cytotoxicity of MCF-7 and 4T1 cancer cells. E The effect of transfection of overexpressed plasmids on the clonogenesis of breast cancer cells was analyzed. F The effect of THC on apoptosis of breast cancer cells transfected with overexpressed plasmid was using the Annexin V/PI dual-labeling technique, and determined by FCM. G The effect of THC on breast cancer cell metastasis after transfection with overexpressed plasmid was analyzed by scratch assay. H Transwell assay was used to analyze the effect of THC on invasion of breast cancer cells after transfection with overexpressed plasmid. Data are expressed as means ± SD from three experiments. (*P < 0.05; **P < 0.01; ***P < 0.001, compared to control; ^#P < 0.05; ^##P < 0.01; ^###P < 0.001, compared to THC)