摘要
目的观察泛素结合酶E2T(UBE2T)基因感染对人卵巢癌细胞株SKOV3增殖的影响,并探讨其机制。方法构建UBE2T⁃shRNA、携带UBE2T、NC⁃shRNA基因的慢病毒载体,将其转染至人卵巢癌细胞株SKOV3,分别记为沉默组、转染组、阴性对照组;并取未转染的人卵巢癌细胞株SKOV3,记为空白对照组,每组设置3个复孔,制成细胞悬液,细胞浓度均调整为1×105个/mL。培养72 h。检测细胞形态、增殖抑制率及AKT等相关蛋白表达。结果沉默组细胞显著减少、连接疏松且有形态改变;沉默组细胞增殖抑制率、G1期细胞占比均高于其余3组(P<0.05),S期和G2期细胞占比、UBE2T、AKT、Cyclin D1 mRNA及蛋白表达、p⁃AKT水平均低于其余3组(P<0.05),转染组细胞增殖抑制率、G1期细胞占比均低于阴性对照组与空白组(P<0.05),S期和G2期细胞占比、UBE2T、蛋白激酶B(AKT)、细胞周期蛋白D1(Cyclin D1)mRNA及蛋白表达、磷酸化⁃AKT(p⁃AKT)水平均高于阴性对照组与空白组(P<0.05)。结论UBE2T表达下调可抑制人卵巢癌细胞株SKOV3增殖,而UBE2T基因过表达则可促进人卵巢癌细胞株SKOV3增殖。
Objective To observe the effect ofubiquitin binding enzyme e2t(UBE2T)geneinfection on the proliferation of human ovarian cancer cell line SKOV3,and to explore its mechanism.Methods UBE2T shRNA and lentiviral vector carrying UBE2T and NC⁃shRNA genes were constructed and transfected into human ovarian cancer cell line SKOV3,andwere recorded as silencing group,transfection group and negative control group,respectively.The untransfected human ovarian cancer cell line SKOV3 was taken and recorded as the blank control group.Three double wells were set in every group to make cell suspension,and the cell concentration was adjusted to 1×105 pieces/mL,culture for 72 h.Results The cells in the silence group were significantly reduced,with loose connection and morphological changes.The inhibition rate of cell proliferation and the proportion of G1 phase cells in the silence group were higher than those in the other three groups(P<0.05),which in the transfection group were lower than those in the negative control group and blank control group(P<0.05),and the percentages of cells in S phase and G2 phase,the mRNA and protein expressions of UBE2T,AKT,Cyclin D1,phosphorylated Akt(p⁃AKT)were lower than those in the other three groups(P<0.05),which in the transfection group were higher than those in the negative control group and blank control group(P<0.05).Conclusion The down⁃regulation of UBE2T expression couldinhibit the proliferation of human ovarian cancer cell line SKOV3,while the overexpres⁃sion of UBE2T gene couldpromote the proliferation of human ovarian cancer cell line SKOV3.
作者
李馨
张瑜
倪婷婷
鲁亮
LI Xin;ZHANG Yu;NI Tingting;LU Liang(Department of Oncology,Guizhou Provincial People′s Hos-pital,Guiyang 550002,China)
出处
《实用医学杂志》
CAS
北大核心
2022年第10期1198-1202,1207,共6页
The Journal of Practical Medicine
基金
贵州省高层次创新型课题(编号:GZSYQCC[2016]003)。
