J Cancer 2020; 11(9):2421-2430. doi:10.7150/jca.37622

Research Paper

FRAT1 Enhances the Proliferation and Tumorigenesis of CD133+Nestin+ Glioma Stem Cells In Vitro and In Vivo

Geng Guo1,*,✉, Jing Liu1,*, Yeqing Ren1,*, Xinggang Mao2,*, Yining Hao1, Chengliang Zhong3, Xiaolin Chen4, Xiaogang Wang1, Yongqiang Wu1, Shizhong Lian1, Lin Mei5, Yuanli Zhao4,✉

1. Department of Neurosurgery, The First Hospital, Shanxi Medical University, No.85 Jiefangnan Road, Taiyuan, Shanxi 030001, People's Republic of China
2. Department of Neurosurgery, Xijing Hospital, Fourth Military Medical University, No.15 Changlexi Road, Xi'an, Shaanxi 710032, People's Republic of China
3. GCP Center, The First Teaching Hospital of Tianjin University of Traditional Chinese Medicine, No.88 Changling Road, Tianjin 300192, People's Republic of China
4. Department of Neurosurgery, Beijing Tiantan Hospital, Capital Medical University, No.6 Tiantan Xili, Beijing 100050, People's Republic of China
5. Department of Orthopedics, The First Hospital, Shanxi Medical University, No.85 Jiefangnan Road, Taiyuan, Shanxi 030001, People's Republic of China
* These authors contributed equally to this work and should be regarded as co-first authors.

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Citation:
Guo G, Liu J, Ren Y, Mao X, Hao Y, Zhong C, Chen X, Wang X, Wu Y, Lian S, Mei L, Zhao Y. FRAT1 Enhances the Proliferation and Tumorigenesis of CD133+Nestin+ Glioma Stem Cells In Vitro and In Vivo. J Cancer 2020; 11(9):2421-2430. doi:10.7150/jca.37622. Available from http://www.jcancer.org/v11p2421.htm

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Abstract

Glioma stem cells (GSCs) are considered the source for development, recurrence, and poor prognosis of glioma, so treatment targeted GSCs is of great interest. The frequently rearranged in advanced T cell lymphomas-1 (FRAT1) gene is an important member of the Wnt/β-catenin signaling transduction pathway, and aberrantly activation of Wnt signaling has been identified to contribute to the tumorigenesis, proliferation, invasion of a variety kinds of cancer stem cells. However, correlations between FRAT1 and GSCs and the specific mechanisms remain unclear. In this study, we aimed to investigate the effect of FRAT1 on GSCs proliferation, colony formation, sphere formation and tumorigenesity in vitro and in vivo and its underlying mechanism. Lentiviral transfection was used to construct GSCs with low FRAT1 expression. The expression of FRAT1 on GSCs proliferation in vitro was assessed by cell counting kit-8(CCK-8). Colony formation and sphere formation assays were conducted to assess the colony and sphere formation ability of GSCs. Then, an intracranial glioma nude mouse model was built to measure the effect of low FRAT1 expression on GSCs proliferation and tumorigenesity in vivo. Real-time PCR, Western blot, and Immunohistochemistry were processed to detect the mRNA and protein expressions of FRAT1, β-catenin in the glioma tissue of xenograft mice to study their correlations. The functional assays verifed that low FRAT1 expression inhibited CD133+Nestin+ GSCs proliferation, colony formation, sphere formation ability in vitro. In vivo GSCs xenograft mice model showed that low FRAT1 expression suppressed the proliferation and tumorigenesity of CD133+Nestin+ GSCs and reduced β-catenin mRNA and protein expression. Furthermore, the expression of FRAT1 and β-catenin were positively correlated. Altogether, results indicate that FRAT1 enhances the proliferation, colony formation, sphere formation and tumorigenesity of CD133+Nestin+ glioma stem cells in vitro and in vivo as well as the expression of β-catenin. Therefore, inhibiting proliferation of GSCs and FRAT1 may be a molecular target to GSCs in treating human glioma in the future.

Keywords: Glioma stem cells (GSCs), Proliferation, The frequently rearranged in advanced T cell lymphomas-1(FRAT1), Wnt/β-catenin pathway