The Mechanism Of Naringin Promoting Gemcitabine Inhibiting Breast Cancer Proliferation And The Construction Of Its Application Model

PurposeTo explore the effect of naringenin on breast cancer and its molecular mechanism,construct breast cancer resistant gemcitabine resistant cell lines and liver metastasis models,and explore the synergistic effect of naringenin and gemcitabine on killing breast cancer cells.Content and method1.The killing effect of naringenin on breast cancer cells(1)The killing effect of naringenin on breast cancer cells and normal breast cell lines was detected by CCK-8,plate clone formation experiment,and flow cytometry;(2)Establish a transplanted tumor model of breast cancer in mice by inoculating tumor cells subcutaneously into mice,and inject intraperitoneally with naringenin to intervene model mice to explore the antitumor effect of naringenin in vivo.(3)Detection of autophagy-related genes by fluorescent quantitative PCR and immunoblotting experiments to explore the effect of naringenin on autophagy of breast cancer cells.2.Gemcitabine-resistant breast cancer cells 4T1 were used to construct nude mice breast cancer liver metastasis model under B superelastic imaging monitoring(1)Continuous low-dose drug treatment to construct gemcitabine-resistant breast cancer cell line 4T1.(2)Gemcitabine-resistant breast cancer cells 4T1 were used to construct nude mice breast cancer liver metastasis model under B-ultrasound monitoring.3.Naringenin can delay gemcitabine resistance(1)Detect the synergistic effect of naringenin and gemcitabine on breast cancer cells by qPCR and cck-8.(2)Detect the killing effect of naringenin and gemcitabine on gemcitabine-resistant breast cancer cell lines by qPCR and cck-8,and explore the effect of naringenin on gemcitabine resistance.Results1.After naringin treatment,the ability of human breast cancer cell line MCF-7 and mouse breast cancer cell line 4T1 cell plate clone decreased(P < 0.001).2.After treatment with naringin,MCF-7 cells and 4T1 cells showed significant apoptosis(P < 0.001).3.After treatment with naringin,the autophagy related genes Beclin1 and Bax increased significantly(P < 0.001),and the autophagy related genes ATG5,HIF-1 α and Bcl-2 increased significantly(P < 0.01).4.Naringin significantly inhibited the increase of tumor volume(P < 0.001)and weight(P < 0.01).5.We successfully induced gemcitabine resistant 4T1 cells(4T1/GEM).IC50(4T1)= 0.0126mg/ml,IC50(4T1 / GEM)= 9.9357mg/ml,RI = 788.547.6.To establish a stable gemcitabine resistant 4T1 cell(4T1/GEM)and a liver metastasis model in nude mice.7.NAR + Gem inhibited 4T1 cells better than NAR(P < 0.05),and NAR + Gem inhibited 4T1 cells better than CTL(P < 0.001).8.The inhibitory effect of NAR + Gem on the proliferation of 4T1/Gem cells was better than that of 4T1 cells(P < 0.05)Conclusions1.Naringenin can inhibit the proliferation of breast cancer cells,and has no obvious or only minor side effects on normal breast cells.Naringenin promotes the death of breast cancer cells through apoptosis and autophagy.2.Naringenin has obvious inhibitory effect on transplanted tumors in mice,and can promote its necrosis;3.Successfully constructed gemcitabine breast cancer resistant cell line 4T1,and successfully constructed its breast cancer liver metastasis model;4.Naringenin can synergize with gemcitabine to kill breast cancer cells,and can delay its resistance to gemcitabine.