miR-200C-3p inhibit the proliferation and migration of prostate cancer cells via targeting ZEB2
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摘要:
目的 前列腺癌的进展与多种因素有关，本研究旨在探索miR-200C-3p对前列腺癌细胞系的影响，并分析miR-200C-3p与ZEB2的潜在机制。 方法 通过qRT-PCR检测miR-200C-3p在前列腺癌细胞系PC-3、DU145、前列腺上皮细胞RWPE-1的表达量。DU145转染miR-200C-3p后，通过CCK-8、划痕修复实验以及Transwell实验探究增殖与迁移能力。沉默ZEB2后分析DU145增殖和迁移能力的变化，采用双荧光素酶报告法蛋白免疫印迹实验测定miR-200C-3p与E盒结合锌指蛋白2(zinc finger E-box binding homeobox 2，ZEB2)的潜在关系。 结果 qRT-PCR结果显示RWPE-1中miR-200C-3p表达量是DU145的2.5倍左右。DU145转染miR-200C-3p mimics后CCK-8实验组的增殖能力明显弱于对照组，划痕修复实验实验组[(20.33±1.45)%]与对照组[(46.67±2.40)%]相比愈合能力明显减弱(P < 0.001)，Transwell实验结果表明实验组(114.30±5.21)与对照组(212.00±6.49)相比迁移能力显著下降(P < 0.001)。沉默ZEB2可抑制DU145细胞的增殖和迁移能力(P < 0.05)，双荧光素酶报告和蛋白免疫印迹实验证实miR-200C-3p在DU145中过表达降低了ZEB2的蛋白表达(P < 0.05)。 结论 miR-200C-3p通过介导ZEB2抑制DU145的增殖和迁移。 -
关键词:
- 前列腺癌 /
- miR-200C-3p /
- E盒结合锌指蛋白2 /
- 增殖 /
- 迁移
Abstract:Objective To explore the effect of miR-200C-3p on prostate cancer cell lines and analyze the potential mechanisms of miR-200C-3p and Zinc finger E-box-binding homeobox 2 (ZEB2). Methods The expression of miR-200C-3p in prostate cancer cell lines PC-3, DU145, and prostate epithelial cells RWPE-1 was detected by qRT-PCR. After DU145 was transfected with miR-200C-3p, the proliferation and migration ability were determined by CCK-8, scratch repair test and Transwell test. After silencing ZEB2, the proliferation and migration ability of DU145 was analyzed, and the potential relationship between miR-200C-3p and ZEB2 was determined by double-luciferase reporter and western blotting. Results The results of qRT-PCR showed that the expression of miR-200C-3p in RWPE-1 was about 2.5 times that of DU145. After DU145 was transfected with miR-200C-3p mimics, the proliferation ability of experimental group was significantly weaker than that of the control group. The healing ability of the experimental group (20.33±1.45) % was significantly lower than that of the control group (46.67±2.40) % (P < 0.001), and the migration ability of the experimental group (114.30±5.21) was significantly lower than that of the control group (212.00±6.49, P < 0.001). Silencing ZEB2 inhibited the proliferation and migration ability of DU145 cells (P < 0.05). The dual luciferase report and western blot experiments confirmed that the overexpression of miR-200C-3p in DU145 reduced the protein expression of ZEB2 (P < 0.05). Conclusion miR-200C-3p inhibits the proliferation and migration of DU145 by mediating ZEB2. -
Key words:
- Prostate cancer /
- miR-200C-3p /
- Zinc finger E-box-binding homeobox 2 /
- Proliferation /
- Migration
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图 2 划痕修复实验和Transwell小室实验检测前列腺癌细胞迁移能力
注：A为2组愈合率比较，aP < 0.05；B为Transwell迁移实验组与对照组比较(×200)。
图 3 沉默ZEB2对DU145细胞增殖和迁移的影响
注：A为沉默ZEB2后CCCK-8检测转染后细胞的增殖能力比较；与si-ZEB2比较，aP < 0.05。B为沉默ZEB2后划痕修复实验组与对照组比较。C为Transwell迁移实验组与对照组比较(×200)。
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