AKR1B10 Promotes Breast Cancer Cell Proliferation And Migration Via PI3K/AKT/NF-κB Signaling Pathway

Background:Breast cancer originates from the mammary gland epithelial tissue.It is the incidence ranks first cancer in female tumors.It accounts for 15%of deaths among tumors in women.The incidence rate is increasing year by year.The greatest threat to women life of breast cancer is not in breast cancer itself but in its metastasis.It is still necessary to further explore the development mechanism of breast cancer,to study new breast cancer diagnosis and treatment methods,and to find new diagnostic or therapeutic targets.Objective:To investigate the association between the expression of AKR1B10 in breast cancer and the prognosis of breast cancer patients,and to explore the effect of AKR1B10 on the proliferation and migration of breast cancer cells.To further explore its mechanism,and to provide a theoretical basis for further study for breast cancer diagnosis or treatment markers.Methods:The association between the high expression of AKR1B10 and the overall prognosis and disease-free survival of breast cancer patients was analyzed by Kaplan-Meier Plotter database.Breast cancer cell lines MCF-7/AKR1B10 stably overexpressed AKR1B10 and breast cancer cell lines BT-20/shAKR1B10 that knockdown AKR1B10 were constructed,respectively.RT-qPCR,Western blot and immunohistochemistry were used to detect the expression of AKR1B10 in breast cancer tissues and breast normal tissues.CCK-8 cell proliferation assay and cell scratch assay were used to detect the proliferation and migration ability of breast cancer cells that overexpressed or knockdown AKR1B10.Western blot was used to detect the expression of proliferation-related proteins cyclinDl,c-myc,survivin and EMT-related proteins twist,Snail,Slug,ZEB1,and E-cadherin in breast cancer cells that overexpressed or knockdown AKR1B10.The PIP3 ELISA kit was used to detect the expression of PIP3 in breast cancer cells that overexpressed or knockdown AKR1B10.Western blot was used to detect the expression of PI3K,p-PI3K,AKT,p-AKT,IKBα,p-IKBα,NF-κB p65,p-NF-κB p65 proteins in breast cancer cells that overexpressed or knockdown AKR1B10;The expression of NF=κB p65 in the nucleus of BT-20 control and BT-20-AKR1B 10-knockdown cells were detected after extracted nuclear protein and plasma protein,respectively.After MCF-7 cells treated with PI3K inhibitor LY94002,western blot was used to detect the PI3K/AKT/NF-κB signal cascade protein expression in total proteins and the expression of NF-κB p65 in nuclear proteins and plasma proteins.Results:The results of the biochemical analysis showed that breast cancer patients with high expression of AKR1B10 have a poor prognosis.After overexpression of AKR1B10,MCF-7 cells had increased proliferation and migration capabilities,and increased expression of cyclinD1,c-myc,survivin and EMT-regulation related proteins while decreased expression of E-cadherin.After knockdown AKR1B10,BT-20 cells had reduced proliferation and migration capabilities,and decreased expression of cyclinDl,c-myc,survivin and EMT-regulation related proteins.PIP3 ELISA assay showed that the expression of PIP3 in MCF-7 cells increased after overexpression of AKR1B10;and the expression of PIP3 in BT-20 cells decreased after AKR1B10 knockdown.The expression of PI3K/AKT/NF-κB signal cascade-related proteins increased after overexpression of AKR1B10 in MCF-7 cells,and the expression of PI3K/AKT/NF-κB signal cascade-related proteins decreased after AKR1B10 knockdown in BT-20 cells.Western bolt showed that the expression of NF-κB p65 in the nucleus of BT-20 cells decreased after AKR1B10 knockdown.The PI3K/AKT/NF-κB signal cascade-related proteins expression in MCF-7 cells decreased after treated with PI3K inhibitor,and western blot showed the nuclear NF-κB p65 expression was decreased after treated with PI3K inhibitor in MCF-7 cells.Conclusion:1.AKR1B10 is upregulated in breast cancer and associated with poor prognosis in breast cancer patients.Patients with high expression of AKR1B10 have a poor prognosis.2.High expression of AKR1B10 promotes breast cancer cell proliferation and migration.3.AKR1B10 promotes the proliferation and migration of breast cancer cells via activating the PI3K/AKT/NF-κB signaling cascade,regulating the expression of proliferation-related and EMT-related proteins.