| 赵 杨,钟山亮,张晓慧,赵建华,李晓平.miR-21靶向调节MARCKS基因影响人乳腺癌细胞多西紫杉醇敏感性的研究[J].现代生物医学进展英文版,2018,(7):1217-1221. |
| miR-21靶向调节MARCKS基因影响人乳腺癌细胞多西紫杉醇敏感性的研究 |
| Effect of MiR-21 Targeting MARCKS Gene on the Sensitivity of Human Breast Cancer Cells to Docetaxel |
| Received:November 03, 2017 Revised:November 28, 2017 |
| DOI：10.13241/j.cnki.pmb.2018.07.004 |
| 中文关键词: 乳腺癌 miR-21 耐药 多西紫杉醇 |
| 英文关键词: Breast cancer miR-21 Chemoresistance Docetaxel |
| 基金项目:国家自然科学基金项目（81272470） |
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| 中文摘要: |
| 摘要 目的：探讨miR-21对MARCKS基因表达的调控及其与乳腺癌MDA-MB-231细胞株多西紫杉醇耐药的关系。方法：通过基因芯片筛选到miR-21在亲代敏感细胞株(MDA-MB-231/S)与多西紫杉醇耐药细胞株(MDA-MB-231/Doc)中存在差异表达，应用实时荧光定量(RT-qPCR)方法验证此差异，同时分别检测MDA-MB-231/S细胞和MDA-MB-231/Doc细胞转染miR-21模拟物和抑制物后的表达变化；通过靶基因预测软件预测miR-21与MARCKS基因的关系；采用四甲基偶氮唑蓝(MTT)法、流式细胞术、RT-qPCR和蛋白质印迹法检测miR-21表达变化对三阴性乳腺癌细胞多西紫杉醇耐药性的影响，并探讨其与MARCKS基因表达的关系。结果：与MDA-MB-231/S相比，miR-21在MDA-MB-231/Doc中的表达水平明显升高(2.03±0.06)倍(P<0.05)。与阴性对照组比，MDA-MB-231/S细胞转染miR-21 mimics后miR-21表达水平明显增高(2.26±0.07)倍(P<0.05)，而MARCKS基因在mRNA和蛋白水平上均明显下调。MDA-MB-231/Doc细胞转染miR-21 inhibitors后，miR-21表达水平是阴性对照组的(0.36±0.03)倍(P<0.05)，MARCKS基因的mRNA和蛋白水平高于其阴性对照组。转染miR-21 mimics后的MDA-MB-231/S细胞IC50显著高于其阴性对照组(P <0.05)，相反，与阴性对照组相比，MDA-MB-231/Doc细胞转染miR-21 inhibitors后，其IC50显著降低(P <0.05)。MDA-MB-231/S转染miR-21 mimics后，miR-21表达量明显上调，MARCKS基因分别是阴性对照组的(3.564±0.336)倍和(2.019±0.268)倍(P<0.05)。结论：miR-21可能通过靶向调节MARCKS基因增强乳腺癌细胞对多西紫杉醇的耐药性。 |
| 英文摘要: |
| ABSTRACT Objective: To investigate the effect of miR-21 on expression of MARCKS，and their influence on chemosensitivity of MDA-MB-231 breast cancer cells to docetaxel. Methods: MicroRNAs microarray and RT-qPCR were conducted to explore the signature of miR-21 between MDA-MB-231/S and MDA-MB-231/Doc. The potential target gene of miR-21 was predicted by online bioinformatic softwares. miR-21 mimics were transfected to MDA-MB-231/S, and inhibitors were transfected to MDA-MB-231/Doc respectively. The IC50 in each group was tested by MTT. The expression of MARCKS was detected by RT-qPCR and Western blot. Results: Compared with control MDA-MB-231/S, the relative expression level of miR-21 in MDA-MB-231/Doc was significantly up-regulated 2.03±0.06 fold, P<0.05. MARCKS is predicted as a target of miR-21 by TargetScan and plays an important role in drug resistance. Drug resistance in mimics-transfected MDA-MB-231/S cells was significantly increased. The expression level of MARCKS was down-regulated. Com- pared with control MDA-MB-231/Doc, drug resistance in inhibitors-transfected MDA-MB-231/Doc cells was significantly decreased. The expression level of MARCKS was up-regulated. The expression of mir-21 was significantly increased after mda-mb-231/S transfec- tion mir-21 mimics, and MARCKS genes were negative control (3.564+0.336) and (2.019+0.268), P<0.05. Conclusion: miR-21 confers breast cancer cells resistant to docetaxel by targeting MARCKS. |
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