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.

/ / Recommend

Add to citation manager EndNote|Ris|BibTeX

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

[1]	Kang Li, Liang Ji, Wei Zhao, Lemin Lin. Dual role of autophagy in biological progression of breast cancer [J]. Chinese Journal of Breast Disease(Electronic Edition), 2023, 17(04): 195-202.
[2]	Changsheng Lin, Jun Zhan, Xue Xiao. Genetic testing and precision molecular targeted therapy in diagnosis and treatment of epithelial ovarian caner [J]. Chinese Journal of Obstetrics & Gynecology and Pediatrics(Electronic Edition), 2023, 19(05): 505-510.
[3]	Dan Luo, Weimin Kong, Shuning Chen, Xiaoling Zhao, Yunkai Xie. Differences in epithelial mesenchymal transition-related biological markers between in-situ and ectopic endometrial epithelial cells in endometriosis patients [J]. Chinese Journal of Obstetrics & Gynecology and Pediatrics(Electronic Edition), 2023, 19(05): 530-539.
[4]	Yunying Zhu, Xiaolin Gao, Yanping Ge, Zhangsong Wang, Zhaoyu Lin, Jinsong Li, Donghui Wu. The expression and role of hypoxia-induced long noncoding RNA LINC00970 in salivary adenoid cystic carcinoma cells [J]. Chinese Journal of Stomatological Research(Electronic Edition), 2023, 17(03): 210-217.
[5]	Fengjin Shang, Luyao Chen, Yaxing Liu, Haoran Zhang, Changhong Lian. Research progress on tumor associated neutrophils in the occurrence, development, and treatment of gastric cancer [J]. Chinese Archives of General Surgery(Electronic Edition), 2024, 18(01): 58-61.
[6]	Hui Zhang, Yi Li, Meiling Dai. Progress in the application of implants in breast reconstruction [J]. Chinese Journal of Operative Procedures of General Surgery(Electronic Edition), 2024, 18(03): 349-352.
[7]	Yongning Li, Xueqin Fu, Ying Li, Peng Liu, Songbai Liu, Yaozhen Pan. Gene similar sequence family member 126A targets vimentin to promote invasion and migration of pancreatic cancer cells and its mechanism [J]. Chinese Journal of Operative Procedures of General Surgery(Electronic Edition), 2023, 17(02): 139-144.
[8]	Qiang Wang, Xin Chen, Xiaoxue Weng, Yongze Zhuang, Guoqing Yu. Indoxyl sulfate induces transdifferentiation of peritoneal mesenchymal cells through TGF-β1 [J]. Chinese Journal of Cell and Stem Cell(Electronic Edition), 2022, 12(06): 329-334.
[9]	Zhuolin Li, Ruxue Jia, Yating Wu, Shenghang Zhang, Shuiliang Wang. Advances in molecular mechanisms underlying tumor metastasis and its targeted intervention [J]. Chinese Journal of Cell and Stem Cell(Electronic Edition), 2022, 12(01): 51-58.
[10]	Zhelong Jiang, Zhihong Wei, Junjie Du, Yi Jiang, Lizhi Lyu, Fang Yang. Effect of FOXC2 on prognosis of liver transplant patients with primary liver cancer [J]. Chinese Journal of Hepatic Surgery(Electronic Edition), 2022, 11(03): 309-314.
[11]	Changhui Geng, Chunling Zhang, Ping Gao, Miao He, Huifeng Li, Yini Yao, Yingxin Liu. LncRNA SNHG17 promotes the metastasis of colorectal cancer by inducing epithelial-mesenchymal transition [J]. Chinese Journal of Colorectal Diseases(Electronic Edition), 2021, 10(04): 367-370.
[12]	Zhiyuan Zhang, Wenwen Li, Shuai Wang, Jiaojiao Gu, Jiaru Gao, Linjiao Jia, Wentao Li. Advancements in application of ultrasound imaging omics in breast tumors [J]. Chinese Journal of Clinicians(Electronic Edition), 2024, 18(01): 87-90.
[13]	Xue Zhang, Miao Yu, Jing Han, Xuhua Hu, Jiayin Liu, Dan Li, Yan Liu, Li Feng, Guiying Wang. Expression of DUOX2 in colon cancer and its effect on malignant behavior of SW620 and HCT116 cells [J]. Chinese Journal of Clinicians(Electronic Edition), 2021, 15(11): 897-903.
[14]	Shihao Li, Yujiao Wang, Zihao Li, Bin Wu, Yinliang Sheng, Yu Qi. Single cell transcriptome analysis of macrophage cap protein on proliferation and metastasis of esophageal squamous cell carcinoma cells [J]. Chinese Journal of Thoracic Surgery(Electronic Edition), 2023, 10(02): 98-105.
[15]	Weichang Ma, Shupeng Zhang. Efficacy of meglumine diatrizoate injection in the treatment of elderly patients with axillary effusion after modified radical mastectomy [J]. Chinese Journal of Geriatrics Research(Electronic Edition), 2020, 07(03): 26-28.

Please choose a citation manager

Content to export