摘要
目的探讨UBE2T对三阴性乳腺癌恶性生物学行为的影响并探讨可能作用机制。方法采用实时荧光定量PCR(QPCR)检测三阴性乳腺癌组织和正常乳腺组织中UBE2T mRNA表达水平。采用GEPIA和Kaplan Meier-plotter在线分析乳腺癌组织和正常乳腺组织中UBE2T表达差异以及UBE2T表达与乳腺癌预后的关系。采用UBE2T shRNA质粒和PCMV-UBE2T质粒转染MDA-MB-231细胞来下调和上调UBE2T表达,采用Western blotting检测验证转染效率。采用CCK-8法、流式细胞术和Transwell小室检测不同UBE2T表达对MDA-MB-231细胞增殖、周期变化、迁移和侵袭的影响。采用Western blotting检测AKT和ERK信号通路蛋白表达。结果三阴性乳腺癌和正常乳腺组织中UBE2T mRNA表达量分别为6.82±1.77和2.12±1.40(P<0.01)。GEPIA和Kaplan Meier-plotter在线分析结果显示乳腺癌中UBE2T高表达,且高表达者OS、RFS较低表达者差,差异有统计学意义(P<0.05)。与对照组比较,上调UBE2T表达后细胞增殖率升高,下调UBE2T表达后细胞增殖率降低,差异有统计学意义(P<0.05)。与对照组比较,PCMV-UBE2T组G1期细胞比率增加,S期细胞比率降低,差异具有统计学意义(P<0.05)。Transwell实验结果显示,sh-UBE2T组迁移和侵袭细胞数分别为266±19、227±14,低于对照组的528±38、406±21,差异具有统计学意义(P<0.05)。Western blotting结果显示,PCMV-UBE2T组p-AKT和p-ERK1/2表达水平分别为1.69±0.14、2.10±0.15,高于对照组(P<0.05)。sh-UBE2T组p-AKT和p-ERK1/2表达水平分别为0.59±0.07、0.35±0.07,低于对照组(P<0.05)。结论UBE2T在乳腺癌组织中高表达,通过ERK和AKT信号途径影响乳腺癌细胞恶性生物学行为。
Objective To explore the effect of UBE2T on malignant biological behavior of triple negative breast cancer and its possible mechanism.Methods Real-time fluorescence quantitative PCR(QPCR)was used to detect the mRNA expression level of UBE2T in triple negative breast cancer tissues and normal breast tissues.GEPIA and Kaplan Meier-plotter were used to analyze the difference of UBE2T expression between breast cancer and normal breast tissue,and the relationship between UBE2T expression and prognosis of breast cancer.UBE2T shRNA and PCMV-UBE2T plasmid were used to transfect MDA-MB-231 cells to down-regulate and up-regulate UBE2T expression,and Western blotting was used to verify the transfection efficiency.CCK-8,flow cytometry and Transwell assay were used to detect the effects of different UBE2T expressions on proliferation,cycle change,migration and invasion of MDA-MB-231 cells.Protein expression of AKT and ERK signaling pathways was detected by Western blotting.Results QPCR results showed that UBE2T mRNA expression in triple negative breast cancer and normal breast tissue were 6.82±1.77 and 2.12±1.40,respectively(P<0.01).The online analysis results of GEPIA and Kaplan Meier-plotter showed that the UBE2T expression in breast cancer was high,and the high expression of UBE2T was lower in OS and RFS,and the difference was statistically significant(P<0.05).Compared with the control group,the cell proliferation rate increased after UBE2T expression was up-regulated,while decreased after UBE2T expression was down-regulated,with statistically significant differences(P<0.05).Compared with the control group,the cell ratio of PCMV-UBE2T group in G1 phase increased,while the cell ratio of S phase decreased significantly,with statistically significant difference(P<0.05).The results of Transwell showed that the number of migrated and invaded cells in sh-UBE2T group was 266±19 and 227±14,respectively,which were lower than those in the control group(528±38 and 406±21,respectively),with statistically significant differences(P<0.05).Western blotting results showed that the expression levels of p-AKT and p-ERK1/2 in PCMV-UBE2T group were 1.69±0.14 and 2.10±0.15,respectively,which were higher than those in the control group(P<0.05).The expression levels of p-AKT and P-ERK1/2 in sh-UBE2T group were 0.59±0.07 and 0.35±0.07,respectively,lower than those in the control group(P<0.05).Conclusion UBE2T is highly expressed in breast cancer tissues and influences the malignant biological behavior of breast cancer cells through ERK and AKT signaling pathways.
作者
吴新军
陈炳合
李帅超
薛明辉
闫争强
WU Xinjun;CHEN Binghe;LI Shuaichao;XUE Minghui;YAN Zhengqiang.(Department of General Surgery, the First Affiliated Hospital of Xinxiang Medical University, Weihui 453100, China)
出处
《临床肿瘤学杂志》
CAS
北大核心
2020年第10期870-875,共6页
Chinese Clinical Oncology
关键词
乳腺癌
UBE2T
增殖
周期
迁移
侵袭
Breast cancer
UBE2T
Proliferation
Cell cycle
Migration
Invision
