Histone deacetylase inhibitor down-regulated the expression of HER-2 in breast cancer through the changes in miRNA
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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，qPCR和Western检测HER-2基因和蛋白水平的变化，同时采用miRNA芯片筛选HDAC抑制剂相关的miRNA谱，qPCR验证miRNA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5 μmol/L组下调93.9%（P < 0.05），而1 μmol/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1 μmol/L时下调92.7%，浓度5 μmol/L时下调87.1%。通过miRNA芯片筛选出7条miRNA，qPCR监测SAHA、TSA处理后，miR-762基因表达上调2.11倍。 结论 HDAC抑制剂可能通过miRNA表达谱改变介导下调乳腺癌HER-2的表达。 Abstract: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. Methods HER-2-positive breast cell lines were treated with HDAC inhibitors. The changes in the gene and protein levels of HER-2 were detected 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 expression of HER-2 in breast cancer cell lines. TSA can down-regulate the expression of HER-2 gene in BT474 and decrease the concentrations 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 5 μmol/L group but not in the 1 μmol/L group. SAHA significantly affected the primary cells at a concentration of 1 μmol/L and reduced the cells at 87.1% at a concentration of 5 μmol/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. -
Key words:
- HDAC inhibitor /
- breast cancer /
- HER-2 /
- miRNA-762
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图 1 HDACi下调乳腺癌细胞中HER-2
Figure 1. HER-2 expression was down-regulated by HDACi in breast cancer cells
A, B. HER-2 expression was down-regulated in breast cancer cells with different concentrations of TSA, *P < 0.05; C, D. HER-2 expression was down-regulated in breast cancer cells with different concentrations of SAHA, *P < 0.05
图 2 HDACi对乳腺癌细胞株HER-2蛋白的影响
Figure 2. Effect of HDACi on HER-2 protein in breast cancer cell lines
图 4 q-PCR验证TSA、SAHA处理BT474细胞48 h后miR-762基因
Figure 4. qPCR was used to verify the miR-762 levels after treating BT474 cells with TSA or SAHA for 48 h
表 1 TSA处理BT474细胞48 h miRNA变化（P < 0.05）
Table 1. MiRNA changes after the BT474 cells were treated with SAHA for 48 h, *P < 0.05
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