摘要
乳腺癌是当今威胁女性健康最主要的恶性肿瘤之一。虽然当前在乳腺癌的诊治方面获得了一定的进展,但其发病率和死亡率仍然较高,寻找有效的乳腺癌预后标记和治疗靶点是当前研究的热点。本研究通过对高通量基因表达数据库(Gene Expression Omnibus,GEO)分析发现,Prominin 2(PROM2)在诱导、增强和增殖成瘤能力的3组MCF7乳腺癌细胞中的表达,高于3组对照组MCF7细胞。且在诱导具有侵袭性的MCF10A细胞中的表达,高于未处理非侵袭性MCF10A细胞。研究选择了人乳腺癌细胞系SK-BR-3,成功地将过表达或敲减PROM2质粒转染到细胞中,并检测PROM2对SK-BR-3细胞的迁移、侵袭和增殖和凋亡的作用。划痕结果显示,敲减PROM2的SKBR-3细胞愈合面积更小(P<0.01),过表达PROM2的SK-BR-3细胞愈合面积更大(P<0.01);Transwell的结果显示,敲减PROM2的SK-BR-3细胞侵袭数量更少(P<0.01),过表达PROM2的SK-BR-3细胞侵袭数量更多(P<0.01);流式细胞术的结果显示,敲减PROM2的SK-BR-3细胞增殖能力减弱且凋亡比率增高(2.50%vs.0.93%),过表达PROM2的SK-BR-3细胞增殖能力增强且凋亡比率降低(0.43%vs.0.72%)。然后,探讨PROM2作用的可能机制。通过基因表达谱交互分析(gene expression profiling interactive analysis,GEPIA),对肿瘤基因图谱数据库(The Cancer Genome Atlas,TCGA)中的乳腺癌数据分析发现,PROM2与β-联蛋白(β-catenin)、人表皮生长因子受体2(human epidermal growth factor receptor 2,HER2)之间表达正相关。通过定量实时聚合酶链反应、免疫荧光和蛋白质印迹也发现,PROM2促进了β-联蛋白和HER2的表达。证明PROM2可能通过激活β-catenin/HER2通路,促进乳腺癌细胞SK-BR-3的迁移、侵袭和增殖,并抑制其凋亡。
Breast cancer is one of the most important malignant tumors that threaten women’s health today.Although some progress has been made in the diagnosis and treatment of breast cancer,its morbidity and mortality are still high.Finding effective breast cancer prognostic markers and treatment targets is a hot spot of current research.In this research,we carried out the Gene Expression Omnibus(GEO)analysis and found that in the three groups of MCF7 breast cancer cells that manifested enhanced proliferation and tumorigenicity,the expression of Prominin 2(PROM2)was higher than the three control groups of MCF7 cells.Moreover,the expression of PROM2 in the induced aggressive MCF10A cells was higher than that in untreated non-invasive MCF10A cells.Then SK-BR-3 breast cancer cells were selected,and the over-expression or knock-down PROM2 plasmid was successfully transfected into the cells,and the effects of PROM2 on the migration,invasion,proliferation and apoptosis of SK-BR-3 cells were detected.The results of scratch assays showed that the healing area of si-PROM2 SK-BR-3 cells was smaller(P<0.01),and hPROM2 SK-BR-3 cells was larger(P<0.01).The results of Transwell assays showed that the invation cell count of si-PROM2 SK-BR-3 cells was less(P<0.01),and hPROM2 SK-BR-3 cells was more(P<0.01).The results of flow cytometry showed that the proliferative capacity of SK-BR-3 cells of the si-PROM2 group was weakened and the apoptotic ratio was enhanced(2.50%vs.0.93%),while the proliferative capacity of the hPROM2 group was increased and the apoptotic ratio was decreased(0.43%vs.0.72%).Then we explored the possible mechanism of PROM2.Analysis of breast cancer data in The Cancer Genome Atlas(TCGA)by gene expression profiling interactive analysis(GEPIA)revealed a positive correlation between PROM2,β-catenin and HER2.Furthermore,the quantitative real-time polymerase chain reaction,immunofluorescence,and Western blotting assays also revealed that PROM2 promoted the expression ofβ-catenin and HER2.We propose that PROM2 may promote the migration,invasion and proliferation of breast cancer SK-BR-3 cells and inhibit its apoptosis by activating theβ-catenin/HER2 pathway.
作者
曾颖靓
厉红元
ZENG Ying-Liang;LI Hong-Yuan(Department of Endocrine and Breast Surgery,The First Affiliated Hospital of Chongqing Medical University,Chongqing 400042,China)
出处
《中国生物化学与分子生物学报》
CAS
CSCD
北大核心
2020年第4期411-420,共10页
Chinese Journal of Biochemistry and Molecular Biology