| 王 敬,邢惠海,徐书方.miR-145靶向Glut1调节Akt通路对乳腺癌细胞增殖和迁移的影响[J].肿瘤学杂志,2020,26(5):418-423. |
| miR-145靶向Glut1调节Akt通路对乳腺癌细胞增殖和迁移的影响 |
| Effect of miR-145 on Proliferation and Migration of Breast Cancer Cells Through Targeting Glut1 to Regulate Akt Pathway |
| 投稿时间：2019-10-29 |
| DOI：10.11735/j.issn.1671-170X.2020.05.B010 |
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| 中文关键词: 微小RNA-145 葡萄糖转运蛋白1 Akt信号通路 乳腺癌 增殖 迁移 |
| 英文关键词:microRNA-145 glucose transporter 1 Akt signaling pathway breast cancer proliferation migration |
| 基金项目:河北省卫生厅科技基金项目（20171007） |
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| 中文摘要: |
| 摘 要：［目的］ 探讨微小RNA-145（miR-145）对葡萄糖转运蛋白1(Glut1)表达和丝苏氨酸激酶(Akt)信号通路的调节作用，以及其对乳腺癌细胞增殖和迁移的影响。［方法］ 2018年2月至2019年2月20例乳腺癌样本及其癌旁组织样本进行Glut1的免疫组化染色，同时采用荧光定量PCR对组织样本中Glut1和miR-145基因表达水平进行检测。采用蛋白免疫印迹实验检测不同乳腺癌细胞株中Glut1蛋白的表达，选择Glut1表达量较高的MDA-MB-231细胞用于后续实验。采用双荧光素酶报告基因实验验证miR-145对Glut1的转录调控，采用磺酰罗丹明B(SRB)实验和细胞迁移侵袭(Transwell)实验验证miR-145对乳腺癌细胞增殖和迁移能力的影响，采用Western blot实验验证Akt信号通路蛋白的表达。［结果］ 与癌旁组织相比，乳腺癌组织中Glut1蛋白和基因表达明显提高（t=7.230，P=0.002），而乳腺癌组织中miR-145表达明显下降（t=4.246，P=0.013）。乳腺癌细胞中转染miR-145后Glut1表达明显下降（t=7.664，P=0.002），并且miR-145对Glut1基因转录具有一定抑制作用，并且抑制效应依赖于Glut1基因的3′-UTR区域。乳腺癌细胞中转染miR-145后细胞在3d和4d的增殖能力（t=7.746和8.086，P=0.001和0.001）以及迁移能力明显下降（t=4.960，P=0.008），并且Akt蛋白的磷酸化水平明显下降（t=5.406，P=0.006）。［结论］ 在乳腺癌中Glut1表达和Akt信号通路的活化能够被miR-145所抑制，并且miR-145能够抑制乳腺癌细胞增殖和迁移。 |
| 英文摘要: |
| Abstract：［Objective］ To investigate the effect of miR-145 on proliferation and migration of breast cancer cells and its mechanism. ［Methods］ Twenty patients with breast cancer admitted in Affiliated Hospital of Hebei University from February 2018 to February 2019 were enrolled. The expression of Glut1 in cancer tissue and pericancerous tissue was detected with immunohistochemy and the expression levels of Glut1 and miR-145 mRNA were detected by fluorescence quantitative RT-PCR. Breast cancer MDA-MB-231 cells were transfeced with miR-145. The transcriptional regulation of miR-145 on Glut1 was verified by double luciferase reporter gene assay，the effects of miR-145 on proliferation and migration of transfected MDA-MB-231 cells were verified by SRB assay and transwell assay，and the expression of Akt signaling protein was verified by Western blot assay. ［Results］ Compared with pericancerous tissues，Glut1 protein and mRNA in breast cancer tissues were significantly increased(t=7.230，P=0.002），while miR-145 expression in breast cancer tissues was significantly decreased（t=4.246，P=0.013）. Glut1 expression was significantly decreased after transfection of miR-145 in MDA-MB-231 cells（t=7.664，P=0.002），and the inhibitory effect was dependent on the 3′-UTR region of Glut1 gene. The proliferation（t=7.746，8.086，P=0.001，0.001）and migration（t=4.960，P=0.008）of breast cancer cells transfected with miR-145 decreased significantly，and the phosphorylation level of Akt also decreased significantly（t=5.406，P=0.006）. ［Conclusion］ Glut1 expression and the activation of Akt signaling pathway can be inhibited by miR-145，leading to inhibiting the proliferation and migration of breast cancer cells. |
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