锌指蛋白ZNF652在乳腺癌组织和细胞中的高表达可促进乳腺癌的发生发展

High expression of ZNF652 promotes carcinogenesis and progression of breast cancer

目的揭示锌指蛋白ZNF652在乳腺癌组织和细胞中的表达水平及其对乳腺癌细胞增殖、侵袭和迁移的影响。方法利用TCGA数据库挖掘ZNF652在乳腺癌组织与癌旁组织中的表达差异,分析ZNF652的表达与分子分型、病理类型、TNM分级和临床分期等临床病理特征的相关性;RT-qPCR和Western blot鉴定ZNF652在MCF-7、MDA-MB-231、SK-BR-3、UACC-812和BT-474乳腺癌细胞系中的表达;通过慢病毒系统构建稳定表达ZNF652的乳腺癌细胞株,利用siRNA敲低ZNF652的表达;通过CCK-8实验和克隆形成实验分析过表达及敲低ZNF652对乳腺癌细胞增殖和集落形成能力的影响;通过Transwell细胞迁移、侵袭实验以及伤口愈合实验阐明过表达及敲低ZNF652对乳腺癌细胞体外迁移和侵袭的影响;采用免疫荧光实验明确ZNF652的亚细胞定位。结果ZNF652在乳腺癌组织中表达显著上调(P<0.001);对于不同乳腺癌分子分型,ZNF652在TNBC型乳腺癌组织中表达下调,而在HER2+型、Luminal A型及Luminal B型乳腺癌组织中表达升高(P<0.01或P<0.001);除粘液癌外,ZNF652在不同病理类型乳腺癌组织中的表达均较癌旁组织升高(P<0.05);高表达ZNF652与乳腺癌远处转移、恶性程度显著相关(P<0.01或P<0.001);ZNF652在5种乳腺癌细胞系中的mRNA和蛋白表达水平均高于正常乳腺细胞(P<0.05或P<0.001);过表达ZNF652促进乳腺癌细胞的增殖、侵袭和迁移,而敲低ZNF652后作用相反;ZNF652定位于细胞核。结论ZNF652在乳腺癌组织和细胞中高表达,促进乳腺癌发生发展,有望作为乳腺癌诊断和治疗的潜在分子靶标。

Objective To investigate the expression of ZNF652 in breast cancer tissues and cells and explore its role in breast cancer cell proliferation,invasion and migration.Methods We exploited the data from the TCGA database to analyze the differential expression of ZNF652 in breast cancer tissues and adjacent tissues and the correlations of ZNF652 expression with the clinicopathological characteristics of breast cancer patients including molecular subtypes,pathological types,TNM stages and clinical stages.RT-qPCR and Western blotting were used to detect the expression of ZNF652 in 5 breast cancer cell lines including MCF-7,MDA-MB-231,SK-BR-3,UACC-812 and BT-474.Using a lentivirus system and siRNA technique,we assessed the effects of ZNF652 over-expression and knockdown on proliferation,colony forming ability,migration and invasion of breast cancer cells with CCK-8 assay,clonogenic assay,Transwell assay and wound healing assay.The subcellular localization of ZNF652 in 293T cells was determined using immunofluorescence assay.Results ZNF652 was significantly up-regulated in breast cancer tissues(P<0.001).In breast cancer tissues of different molecular types,ZNF652 was down-regulated in TNBC breast cancer tissues but increased in HER2+,Luminal A and Luminal B breast cancer tissues(P<0.01 or 0.001).The expression of ZNF652 was significantly higher in breast cancer tissues of all pathological types except for mucinous carcinoma than in the adjacent tissues(P<0.05).The high expression of ZNF652 was closely related to distant metastasis and malignancy of breast cancer(P<0.01 or 0.001).The mRNA and protein expression levels of ZNF652 was significantly higher in the 5 breast cancer cell lines than in normal breast cells(P<0.05 or 0.001).Overexpression of ZNF652 promoted the proliferation,invasion and migration of breast cancer cells,while ZNF652 knockdown produced the opposite effects(P<0.05).Immunofluorescence assay identified subcellular localization of ZNF652 in the nuclei of 293T cells.Conclusion ZNF652 is highly expressed in breast cancer tissues and cells to promote the development and progression of breast cancer and may serve as a potential molecular target for diagnosis and treatment of the malignancy.