FOXC2 affects the invasion and migration ability of breast cancer by Hedgehog/Gli signaling pathway

doi:10.3877/cma.j.issn.1674-0793.2020.05.002

Abstract

Abstract:

Objective

To explore the mechanism of FOXC2 mediated Hedgehog/Gli signaling pathway involved in breast cancer invasion and migration by regulating epithelium mesenchymal transformation (EMT).

Methods

Breast cancer MDA-MB-231 cells were treated with different concentrations Cyclopamine-a Hedgehog/Gli signaling pathway inhibitor. MTT method was used to detect the cell proliferation inhibition rate and calculated the drug half inhibitory concentration (IC₅₀). Cyclopamine or silencing FOXC2 expression by RNA interference (siRNA) was used to block the Hedgehog/Gli signaling pathway activation. Transwell chamber invasion assay and scratch assay were used to detect invasion and migration ability of MDA-MB-231 cells after blocking Hedgehog/Gli signal pathway respectively. Western blotting was used to detect the expression of Hedgehog/Gli signaling pathway molecules Smoothened (Smo), Gli1 and EMT-related markers E-cadherin and Vimentin protein before and after blocking Hedgehog/Gli signaling pathway.

Results

Compared with the blank control group, Cyclopamine could significantly inhibit the MDA-MB-231 cells proliferation and showed a time-dose effect relationship. The half inhibition inhibitory concentration was 25 μmol/L after 48 hour. Transwell chamber invasion assay and wound healing assay showed that FOXC2-siRNA group and Cyclopamine group penetrating cells and migration rates were significantly lower, as compared with the untransfected group and Control-siRNA group. The difference was statistically significant (P<0.05). Western blotting results also showed that the expression of Smo, Gli1 and FOXC2 protein was significantly decreased in FOXC2-siRNA group and Cyclopamine group, as compared with the untransfected group and Control-siRNA group (P<0.05). The expression of Vimentin protein was significantly decreased and the expression of E-cadherin protein was increased when silencing FOXC2 expression, the differences were statistically significant (P<0.05).

Conclusions

FOXC2 mediates Hedgehog/Gli signaling pathway involved in breast cancer invasion and migration by regulating EMT. It is suggested that blocking the Hedgehog/Gli signal pathway may be a new targeted therapy of breast cancer.

Key words: FOXC2, Hedgehog/Gli signal pathway, Epithelial-mesenchymal transition, Breast tumor, Invasion and metastasis

Hongbo Qu, Dixian Luo, Lili Duan, Xiongqiang Hu, Chengyi Wu. FOXC2 affects the invasion and migration ability of breast cancer by Hedgehog/Gli signaling pathway[J]. Chinese Archives of General Surgery(Electronic Edition), 2020, 14(05): 326-330.

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URL: https://zhptwkxwx.cma-cmc.com.cn/EN/10.3877/cma.j.issn.1674-0793.2020.05.002

https://zhptwkxwx.cma-cmc.com.cn/EN/Y2020/V14/I05/326

Figures/Tables 5

References 11

[1]	Pietrobono S, Gagliardi S, Stecca B. Non-canonical Hedgehog signaling pathway in cancer: activation of GLI transcription factors beyond Smoothened[J]. Front Genet, 2019, 10: 556.
[2]	周明莉, 唐石伏, 张海龙, 等. Hedgehog信号通路调控Twist高表达乳腺肿瘤干细胞样细胞的富集及迁移[J].肿瘤，2017, 37(1): 1024-1031.
[3]	Neelakantan D, Zhou H, Oliphant M, et al. Publisher Correction: EMT cells increase breast cancer metastasis via paracrine GLI activation in neighbouring tumour cells[J]. Nat Commun, 2018, 9(1): 4720.
[4]	Yang H, Chen T, Xu S, et al. Long noncoding RNA FOXC2-AS1 predicts poor survival in breast cancer patients and promotes cell proliferation[J]. Oncol Res, 2019, 27(2): 219-226.
[5]	Pandolfi S, Stecca B. Cooperative integration between HEDGEHOG-GLI signalling and other oncogenic pathways: implications for cancer therapy[J]. Expert Rev Mol Med, 2015, 17: e5.
[6]	屈洪波, 吴诚义, 范原铭, 等. 沉默FOXC2对TGF-β1诱导的MCF-7细胞上皮-间质转化的逆转作用[J]. 中国病理生理杂志, 2013, 29(5): 850-856.
[7]	Mani SA, Yang J, Brooks M, et al. Mesenchyme Forkhead 1 (FOXC2) plays a key role in metastasis and is associated with aggressive basal-like breast cancers[J]. Proc Natl Acad Sci U S A, 2007, 104(24): 10069-10074.
[8]	Cai J, Tian AX, Wang QS, et al. FOXF2 suppresses the FOXC2-mediated epithelial-mesenchymal transition and multidrug resistance of basal-like breast cancer[J]. Cancer Lett, 2015, 367(2): 129-137.
[9]	Li Y, Yang W, Yang Q, et al. Nuclear localization of GLI1 and elevated expression of FOXC2 in breast cancer is associated with the basal-like phenotype[J]. Histol Histopathol, 2012, 27(4): 475-484.
[10]	Wang L, Li M, Zhou Y, et al. MicroRNA Let-7g directly targets forkhead box C2 (FOXC2) to modulate bone metastasis in breast cancer[J]. Open Med (Wars), 2017, 12: 157-162.
[11]	Pham T, Perez White BE, Zhao H, et al. Protein kinase C α enhances migration of breast cancer cells through FOXC2-mediated repression of p120-catenin[J]. BMC Cancer, 2017, 17(1): 832.

组别	24 h	48 h	72 h
空白对照组	10.32±1.13	10.47±1.25	11.62±1.04
低浓度组(10.0 μmol/L)	12.39±1.43	15.35±1.72	16.03±1.07
中浓度组(20.0 μmol/L)	15.24±1.26	45.08±2.17^a	71.25±2.21^a
高浓度组(40.0 μmol/L)	21.18±1.37^a	64.15±3.24^a	87.37±3.43^a

组别	24 h	48 h	72 h
空白对照组	10.32±1.13	10.47±1.25	11.62±1.04
低浓度组(10.0 μmol/L)	12.39±1.43	15.35±1.72	16.03±1.07
中浓度组(20.0 μmol/L)	15.24±1.26	45.08±2.17^a	71.25±2.21^a
高浓度组(40.0 μmol/L)	21.18±1.37^a	64.15±3.24^a	87.37±3.43^a

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