TET1蛋白对人乳腺癌MDA-MB-231细胞株增殖和侵袭能力的影响及其相关机制

发布时间：2018-06-22 02:35

本文选题：TET + 乳腺癌　；参考：《肿瘤防治研究》2017年07期

【摘要】：目的探讨TET1对乳腺癌MDA-MB-231细胞增殖、转移、侵袭等生物学行为的影响,并初步探究其相关分子机制。方法应用慢病毒载体建立TET1过表达稳转细胞株(MDA-MB-231-TET1)及阴性对照组稳转细胞株(MDA-MB-231-NC),Real-time PCR及Western blot法检测TET1的表达情况。CCK-8法、流式细胞术检测细胞的增殖及细胞周期分布,细胞划痕实验及Transwell小室检测细胞迁移及侵袭能力;Western blot及免疫荧光实验检测EMT相关蛋白(E-cadherin、N-cadherin、Vimentin、β-catenin)分子的表达。结果成功构建过表达TET1细胞株(P=0.03)。相较于阴性对照组和空白对照组细胞,MDA-MB-231-TET1组细胞增殖、迁移及侵袭明显受到抑制(P0.001),G2/M期细胞周期阻滞(P=0.002);同时,伴随E-cadherin表达升高(P0.001),N-cadherin(P=0.003)、Vimentin(P=0.041)表达降低,β-catenin(P0.001)表达降低,并且TET1可抑制β-catenin向细胞核内转移,进而抑制EMT的发生。结论 TET1可抑制MDA-MB-231细胞增殖,并能抑制其迁移及侵袭,其机制可能与通过Wnt/β-catenin通路抑制EMT的发生相关。
[Abstract]:Objective to investigate the effects of TET1 on proliferation, metastasis and invasion of breast cancer MDA-MB-231 cells, and to explore its molecular mechanism. Methods the expression of TET1 was detected by real-time PCR and Western blot, and the proliferation and cell cycle distribution were detected by flow cytometry (FCM), and the expression of Tet 1 was detected by real-time PCR and Western blot assay respectively by using lentivirus vector and MDA-MB-231-TET1 and MDA-MB-231-NC, respectively. The expression of E-cadherin N-cadherin (尾 -catenin) was detected by cell scratch assay, transwell chamber assay, and the expression of E-cadherin N-cadherin (尾 -catenin) by Western blot. Results TET1 cell line was successfully constructed (P0. 03). The cell proliferation, migration and invasion of MDA-MB-231-TET1 cells were significantly inhibited (P0.001) in G _ 2 / M phase cell cycle arrest (P _ (0.002), and the expression of 尾 -catenin (P _ (0.001) decreased with the increase of E-cadherin expression (P _ (0.001), N-cadherin (P _ (0.003) Vimentin (P _ (0.041) expression decreased, and the expression of 尾 -catenin (P _ (0.001) decreased in MDA-MB-231-TET1 group. Moreover, TET1 inhibited 尾-catenin transfer to the nucleus, which inhibited the occurrence of EMT. Conclusion TET1 can inhibit the proliferation, migration and invasion of MDA-MB-231 cells, and its mechanism may be related to the inhibition of EMT through Wnt- 尾 -catenin pathway.
【作者单位】：河北医科大学第四医院乳腺中心;
【基金】：河北省自然科学基金(H2015206466)
【分类号】：R737.9

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