摘要
目的:探讨组蛋白去乙酰化酶(histone deacetylase,HDAC)抑制剂下调乳腺癌HER-2的表达机制,为乳腺癌抗HER-2治疗提供新的实验依据。方法:利用HDAC抑制剂处理HER-2阳性乳腺细胞,q PCR和Western检测HER-2基因和蛋白水平的变化,同时采用mi RNA芯片筛选HDAC抑制剂相关的mi RNA谱,q PCR验证mi RNA表达变化。结果:体外细胞实验证实HDAC抑制剂TSA和SAHA可下调乳腺癌细胞系HER-2的表达,TSA可下调BT474的HER-2基因表达,浓度为100 nmol时下调10.7%,浓度为200 nmol时下调38.9%(P<0.05)。TSA对原代细胞HER-2基因表达无明显下调(P>0.05)。SAHA对BT474中HER-2基因表达的影响,浓度5μmol/L组下调93.9%(P<0.05),而1μmol/L组无明显下调。SAHA对原代细胞HER-2基因表达下调较为明显,浓度1μmol/L时下调92.7%,浓度5μmol/L时下调87.1%。通过mi RNA芯片筛选出7条mi RNA,q PCR监测SAHA、TSA处理后,mi R-762基因表达上调2.11倍。结论:HDAC抑制剂可能通过mi RNA表达谱改变介导下调乳腺癌HER-2的表达。
Objective:To investigate the mechanism of histone deacetylase (HDAC) inhibitor in down-regulating the expression of HER-2 in breast cancer cells and to provide an innovative therapeutic option to overcome the disadvantages of anti-HER-2 therapy. Meth-ods:HER-2-positive breast cell lines were treated with HDAC inhibitors. The changes in the gene and protein levels of HER-2 were de-tected by qPCR and Western blot. MiRNA microarray was used to identify the HDAC inhibitors, whereas qPCR was used to verify the miRNA expression. Results:In vitro cell experiments confirmed that the HDAC inhibitors TSA and SAHA can down-regulate the expres-sion of HER-2 in breast cancer cell lines. TSA can down-regulate the expression of HER-2 gene in BT474 and decrease the concentra-tions of 100 nmol by 10.7%and 200 nmol by 38.9%(P〈0.05). TSA had no effect on the primary cells. The expression of HER-2 gene of BT474 was down-regulated by 93.9%(P〈0.05) in the 5μmol/L group but not in the 1μmol/L group. SAHA significantly affected the pri-mary cells at a concentration of 1μmol/L and reduced the cells at 87.1%at a concentration of 5μmol/L. Seven miRNAs were identified from the miRNA microarray. MiR-762 was used as a basis to identify the changes in miRNA. The miRNA sputum identified by miRNA microarray and qPCR may be associated with the down-regulation of HER-2 by HDAC inhibitors. Conclusion: HDAC inhibitors may down-regulate the expression of HER-2 in breast cancer cells by changing some miRNAs.
出处
《中国肿瘤临床》
CAS
CSCD
北大核心
2017年第13期644-648,共5页
Chinese Journal of Clinical Oncology
基金
国家自然科学基金面上项目(编号:81472183)
国家科技支撑计划课题(编号:2015BAI12B15)资助~~