Notch3调控Hes2逆转三阴性乳腺癌上皮-间质转化的机制研究

目的 探究Notch3逆转三阴性乳腺癌（TNBC）上皮-间质转化（EMT）及抑制肿瘤转移的分子机制。方法 将Notch3过表达质粒（pcDNA3.1-Notch3-GFP）及其阴性对照质粒（pcDNA3.1-NC-GFP）、Hes2敲低的小分子干扰质粒（si-Hes2-GFP）及其阴性对照质粒（si-NC-GFP）转染...

The mechanism of Notch3 regulating Hes2 to reverse epithelial-mesenchymal transition in triple-negative breast cancer

Objective To explore the molecular mechanism of Notch3 reversing epithelial-mesenchymal transition (EMT) and inhibiting tumor metastasis in triple negative breast cancer (TNBC). Methods The Notch3 overexpression plasmid (pcDNA3.1-Notch3-GFP) and its negative control plasmid (pcDNA3.1-NC-GFP), the small molecule interference plasmid for Hes2 knockdown (si-Hes2-GFP) and its negative control plasmid (si-NC-GFP) were transfected into TNBC MDA-MB-231 cells. Cells were divided into the control group (no transfection was performed), the pc-NC group (transfection pcDNA3.1-NC-GFP), the Notch3 overexpression group (pc-Notch3 group, transfection pcDNA3.1-Notch3-GFP), the Notch3 overexpression+si-NC group (pc-Notch3+si-NC group, co-transfected pcDNA3.1-Notch3-GFP and si-NC-GFP), and the Notch3 overexpression+Hes2 knockdown group (pc-Notch3+si-Hes2 group, co-transfected pcDNA3.1-Notch3-GFP and si-Hes2-GFP). Real-time quantitative PCR and Western blot assay were performed to measure mRNA and protein expression levels of Notch3 and Hes2 in cells to verify the transfection efficiency. The cell counting kit-8 assay was performed to measure cell proliferation activity. Scratch healing assay and Transwell chamber assay were performed to measure cell migration and invasion abilities. Western blot assay was performed to measure EMT-related proteins (E-cadherin and Vimentin) in cells. Results Compared with the control group and the pc-NC group, the Notch3, Hes2 mRNA and protein, E-cadherin protein levels were significantly increased in the pc-Notch3 group and the pc-Notch3+si-NC group, and the proliferation activity, mobility, number of transmembrane cells and Vimentin protein levels were significantly decreased (P<0.05). Compared with the pc-Notch3 group and the pc-Notch3+si-NC group, the Hes2 mRNA and protein levels and E-cadherin protein were significantly decreased in the pc-Notch3+si-Hes2 group, and the proliferation activity, mobility, number of transmembrane cells and Vimentin protein levels were significantly increased (P<0.05). Conclusion Notch3 may reverse the EMT of TNBC and inhibit tumor metastasis by up-regulating Hes2.