Study On The Mechanism That Estradiol Regulated NgBR Protein To Induce Cervical Cancer Cell Proliferation And Migration Via Wnt/β-cat Pathway

Objective:Taking cervical cancer cell lines and genetically modified cervical cancer cells cultured in vitro as the research objects,explore the effect of estradiol（E2）on the expression of NgBR via the Wnt/β-cat signaling pathway to further affect the proliferation,migration and invasion of cervical cancer cells.Methods:1)Taking cervical cancer cell lines cultured in vitro as the research object,observe the effect of estradiol（E2）on its proliferation,migration and invasion,and clarify the role and mechanism of NgBR in it.1.1 Taking the cervical cancer cell line（Hela cell line）cultured in vitro as the research object,use different concentrations of estradiol(0,1×10^-11,1×10^-10,1×10^-9,1×10^-8 M)to culture it.1.2 Use Western blotting technology to detect the expression of NgBR,β-catenin,k-ras and c-myc in each group of cell lines,use MTT method to detect cell proliferation activity,and use Transwell method and scratch experiment to observe cell migration and invasion in each group.1.3 Explore the effect of estradiol（E2）on the expression level of NgBR in cervical cancer cells,and further explain the relationship between the expression level of NgBR protein and the growth,proliferation,and migration of cervical cancer cells.2)Taking genetically modified cervical cancer cells as the research object,confirm that NgBR affects the expression of k-ras and c-myc via Wnt/β-cat signaling pathway,and further affects the proliferation,migration and invasion of cervical cancer cells.2.1 Use gene transfection technology to silence the NUS1P3 gene corresponding to NgBR;2.2 Use Western blotting technology to detect the expression levels of NgBR,β-catenin,k-ras,and c-myc protein in cells in the control group（NC group）and silent group（Si-NgBR group）.2.3 Confirm that NgBR affects the expression of k-ras and c-myc via Wnt/β-cat signaling pathway,and further affects the proliferation,migration and invasion of cervical cancer cells.Results:1)Compared with estradiol 0M group,the expression of NgBR、β-catenin、k-ras、c-myc increased gradually with the increase of estradiol concentration（P<0.05）;2)Compared with estradiol 0M group,the activity of Hela cells increased gradually with the increase of estradiol concentration（P<0.05）;3)Compared with estradiol 0M group,with the increase of estradiol concentration,the invasion ability of Hela cells increased gradually（P<0.05）;4)Compared with estradiol 0M group,with the increase of estradiol concentration,the migration ability of Hela cells increased gradually（P<0.05）;5)Compared with NC group,the expression of NgBR、β-catenin、k-ras、c-myc in si-NgBR group decreased（P<0.05）,and NgBR had a positive correlation withβ-catenin、k-ras、c-myc protein expression（P<0.05）.Conclusion:Estradiol may act on the NgBR protein and increase its expression level,thereby increases the proliferation、migration and invasion ability of cervical cancer cells.The mechanism may be that NgBR up-regulates the expression ofβ-catenin,k-ras and c-myc via the Wnt/β-cat pathway.