TGFβ 调节 NK1R-Tr 影响乳腺癌细胞的增殖和凋亡。
TGFβ regulates NK1R-Tr to affect the proliferation and apoptosis of breast cancer cells.
机构信息
Department of Clinical Laboratory, Tianjin Medical University Cancer Institute and Hospital, National Clinical Research Center for Cancer, Tianjin's Clinical Research Center for Cancer, Tianjin Key Laboratory of Cancer Prevention and Therapy, Key Laboratory of Breast Cancer Prevention and Therapy of Educational Ministry, Tianjin Medical University, Tianjin, China; Department of Clinical Laboratory, Tianjin Children's Hospital, Tianjin, China.
Department of Clinical Laboratory, Tianjin Medical University Cancer Institute and Hospital, National Clinical Research Center for Cancer, Tianjin's Clinical Research Center for Cancer, Tianjin Key Laboratory of Cancer Prevention and Therapy, Key Laboratory of Breast Cancer Prevention and Therapy of Educational Ministry, Tianjin Medical University, Tianjin, China.
出版信息
Life Sci. 2020 Sep 1;256:117674. doi: 10.1016/j.lfs.2020.117674. Epub 2020 May 5.
OBJECTIVES
TGFβ promotes cancer aggressiveness in advanced stages. NK1R-Tr expression in advanced breast cancer has a pro-carcinogenic effect. In this study, we aimed to investigate the effect of the association of TGFβ with NK1R-Tr expression on the proliferation and apoptosis of breast cancer cells.
METHODS
Immunohistochemical staining and Western blot analysis were used to detect TGFβ and NK1R-Tr in breast cancer and paracancerous tissue samples. MDA-MB-231 and BT549 cells were stimulated with TGFβ after NK1R knockdown or treated with the NK1R antagonist aprepitant, and the effects of TGFβ and NK1R-Tr on proliferation and apoptosis were detected by CCK-8, colony formation and flow cytometry assays. In vivo xenograft models were used to further verify the effects of NK1R-Tr and TGFβ. The regulatory effects of Smad4 on NK1R promoter activity were confirmed by ChIP and dual-luciferase reporter assays.
RESULTS
The expression levels of TGFβ and NK1R-Tr were higher in breast cancer tissues than in adjacent tissues and were positively correlated in human breast cancer tissues. NK1R knockdown or aprepitant treatment in MDA-MB-231 and BT549 cells attenuated the effects of TGFβ on cell proliferation. The proportion of cells in G2/M phase significantly increased, the expression of cyclin B1 decreased, and the expression of P21 increased; these effects were weakened by TGFβ treatment. Apoptosis in breast cancer cells was significantly increased. In vivo xenograft models were used to further verify that NK1R-Tr and TGFβ promoted tumour growth. After TGFβ treatment, the binding capacity of Smad4 to the NK1R promoter, as well as luciferase activity, was enhanced.
CONCLUSIONS
The expression levels of TGFβ and NK1R-Tr were higher in breast cancer tissues than in normal tissues, and both were correlated with a poor patient prognosis. TGFβ and NK1R-Tr promoted cell proliferation and inhibited apoptosis, and TGFβ regulated the expression of NK1R-Tr via Smad4.
目的
TGFβ 在晚期促进癌症侵袭性。NK1R-Tr 在晚期乳腺癌中的表达具有致癌作用。本研究旨在探讨 TGFβ 与 NK1R-Tr 表达的关联对乳腺癌细胞增殖和凋亡的影响。
方法
免疫组化染色和 Western blot 分析用于检测乳腺癌和癌旁组织样本中的 TGFβ 和 NK1R-Tr。在 NK1R 敲低后，用 TGFβ 刺激 MDA-MB-231 和 BT549 细胞，并用 CCK-8、集落形成和流式细胞术检测 TGFβ 和 NK1R-Tr 对增殖和凋亡的影响。体内异种移植模型进一步验证了 NK1R-Tr 和 TGFβ 的作用。用 ChIP 和双荧光素酶报告基因检测证实了 Smad4 对 NK1R 启动子活性的调节作用。
结果
TGFβ 和 NK1R-Tr 在乳腺癌组织中的表达水平高于相邻组织，且在人乳腺癌组织中呈正相关。在 MDA-MB-231 和 BT549 细胞中，NK1R 敲低或 aprepitant 处理减弱了 TGFβ 对细胞增殖的影响。G2/M 期细胞比例显著增加，cyclin B1 表达减少，P21 表达增加；TGFβ 处理减弱了这些作用。乳腺癌细胞凋亡明显增加。体内异种移植模型进一步验证了 NK1R-Tr 和 TGFβ 促进肿瘤生长。TGFβ 处理后，Smad4 与 NK1R 启动子的结合能力以及荧光素酶活性增强。
结论
TGFβ 和 NK1R-Tr 在乳腺癌组织中的表达水平高于正常组织，且均与患者预后不良相关。TGFβ 和 NK1R-Tr 促进细胞增殖，抑制细胞凋亡，TGFβ 通过 Smad4 调节 NK1R-Tr 的表达。
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