MicroRNA-452 contributes to the docetaxel resistance of breast cancer cells
MicroRNA-452 contributes to the docetaxel resistance of breast cancer cells
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DOI：
10.1007/s13277-014-1834-z
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发表时间：
2014-07-01
期刊：
影响因子：
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通讯作者：
Zhao, Jian-hua
中科院分区：
文献类型：
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作者：
Hu, Qing;Chen, Wei-xian;Zhao, Jian-hua
MicroRNA-452（miRNA-452）在耐紫杉醇的人乳腺癌MCF-7细胞（MCF-7/DOC）中过表达。然而，其在调节乳腺癌细胞对多西他赛（DOC）的敏感性中的作用仍不清楚。为探讨miRNA-452在乳腺癌细胞对DOC敏感性中的作用，采用实时荧光定量PCR（RT-qPCR）技术检测MCF-7/DOC和MCF-7细胞中miRNA-452的差异表达。将miRNA-452模拟物转染到MCF-7细胞中，并将miRNA-452抑制剂转染到MCF-7/DOC细胞中。采用RT-qPCR、MTT法和流式细胞术检测miRNA-452在转染细胞中的作用。通过RT-qPCR和Western blot分析miRNA-452与其预测靶基因"后期促进复合物4"（APC4）之间的关系，结果发现miRNA-452在MCF-7/DOC细胞中的表达量是亲本MCF-7细胞的78.9倍。在模拟转染的MCF-7细胞中，miRNA-452的表达上调了212.2倍DOC的半数抑制浓度（IC_（50））与阴性对照组（NC）相比有显著性差异（P <0.05（1.98 ± 0.15 μ M）显著高于NC（0.85 ± 0.08 μ M，P <0.05）或空白对照（1.01 ± 0.19 μ M，P <0.05）。细胞凋亡率（6.3 ± 1.3%）明显低于正常对照组（23.8 ± 6.6%）和空白对照组（18.6 ± 4.7%）（P <0.05）。相比之下，在转染了cDNAor的MCF-7/DOC细胞中，miRNA-452的表达下调了0.58倍DOC的IC_（50）值与NC相比，差异有统计学意义（P <0.05）。（44.5 +/-3.2 μ M）显著低于其NC（107.3 +/-6.63 μ M，P <0.05）或空白对照（102.22 ± 11.34 μ M，P <0.05），凋亡率（45.5 ± 10.8%）明显高于NC（9.9 ± 2.2%，P <0.05）和空白对照（9.4 ± 2.5%，P <0.05）。此外，体外研究发现，乳腺癌细胞中miRNA-452与APC 4的表达呈负相关，miRNA-452的失调可能部分通过靶向APC 4参与乳腺癌细胞DOC耐药的形成。
MicroRNA-452 (miRNA-452) was overexpressed in docetaxel-resistant human breast cancer MCF-7 cells (MCF-7/DOC). However, its role in modulating the sensitivity of breast cancer cells to docetaxel (DOC) remains unclear. The aim of this study is to investigate the role of miRNA-452 in the sensitivity of breast cancer cells to DOC.Real-time quantitative PCR (RT-qPCR) were used to identify the differential expression of miRNA-452 between MCF-7/DOC and MCF-7 cells. MiRNA-452 mimic was transfected into MCF-7 cells and miRNA-452 inhibitor was transfected into MCF-7/DOC cells. The role of miRNA-452 in these transfected cells was evaluated using RT-qPCR, MTT assay, and flow cytometry assay. The relationship of miRNA-452 and its predictive target gene "anaphase-promoting complex 4" (APC4) was analyzed by RT-qPCR and Western blot.MiRNA-452 showed significantly higher expression (78.9-folds) in MCF-7/DOC cells compared to parental MCF-7 cells. The expression of miRNA-452 in the mimic transfected MCF-7 cells was upregulated 212.2-folds (P < 0.05) compared to its negative control (NC), and the half maximal inhibitory concentration (IC50) value of DOC (1.98 +/- 0.15 mu M) was significantly higher than that in its NC (0.85 +/- 0.08 mu M, P < 0.05) or blank control (1.01 +/- 0.19 mu M, P < 0.05). Furthermore, its apoptotic rate (6.3 +/- 1.3 %) was distinctly decreased compared with that in its NC (23.8 +/- 6.6 %, P < 0.05) or blank control (18.6 +/- 4.7 %, P < 0.05). In contrast, the expression of miRNA-452 in the inhibitor-transfected MCF-7/DOC cells was downregulated 0.58-fold (P < 0.05) compared to its NC, the IC50 value of DOC (44.5 +/- 3.2 mu M) was significantly lower than that in its NC (107.3 +/- 6.63 mu M, P < 0.05) or blank control (102.22 +/- 11.34 mu M, P < 0.05), and the apoptotic rate (45.5 +/- 10.8 %) was distinctly increased compared with its NC (9.9 +/- 2.2 %, P < 0.05) and blank control (9.4 +/- 2.5 %, P < 0.05). Further, there was an inverse association between miRNA-452 and APC4 expression in breast cancer cells in vitro.Dysregulation of miRNA-452 involved in the DOC resistance formation of breast cancer cells may be, in part, via targeting APC4.