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Inhibition of the CyclinD1 promoter in response to sonic hedgehog signaling pathway transduction is mediated by Gli1

Medulloblastoma (MB) is the most common malignant tumor of the central nervous system in children. Accumulating evidence suggests a major role for the activation of the sonic hedgehog (SHH) signaling pathway in the development of MB cells; however, the mechanisms underlying the effect of this pathwa...

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Autores principales: Lin, Zhongxiao, Sheng, Hansong, You, Chaoguo, Cai, Ming, Zhang, Yiping, Yu, Li Sheng, Yu, Xiaoming, Lin, Jian, Zhang, Nu
Formato: Online Artículo Texto
Lenguaje:English
Publicado: D.A. Spandidos 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5244851/
https://www.ncbi.nlm.nih.gov/pubmed/28123507
http://dx.doi.org/10.3892/etm.2016.3969
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author Lin, Zhongxiao
Sheng, Hansong
You, Chaoguo
Cai, Ming
Zhang, Yiping
Yu, Li Sheng
Yu, Xiaoming
Lin, Jian
Zhang, Nu
author_facet Lin, Zhongxiao
Sheng, Hansong
You, Chaoguo
Cai, Ming
Zhang, Yiping
Yu, Li Sheng
Yu, Xiaoming
Lin, Jian
Zhang, Nu
author_sort Lin, Zhongxiao
collection PubMed
description Medulloblastoma (MB) is the most common malignant tumor of the central nervous system in children. Accumulating evidence suggests a major role for the activation of the sonic hedgehog (SHH) signaling pathway in the development of MB cells; however, the mechanisms underlying the effect of this pathway on tumor survival and growth remain poorly understood. The Gli family zinc finger 1 (Gli1) transcription factor is considered as a mediator of the SHH signaling pathway in MB cells. Therefore, the present study investigated whether the SHH signaling pathway promotes the apoptosis of MB cells via downregulation of Gli1. GANT61, a novel Gli1 inhibitor, is known to have an in vitro activity against tumors. In the current study, Daoy cells were treated with different concentrations of GANT61 for 24 h, and the effect on cell proliferation was assayed by cell counting kit-8 assay. In addition, the cell cycle progression and apoptosis were assayed by flow cytometry analysis and hematoxylin-eosin (HE) staining. The effects of GANT61 treatment on SHH signaling pathway at the mRNA level were assayed by polymerase chain reaction (PCR). To further elucidate the inhibitory effects of GANT61 on the expression of Gli1 and CyclinD1, their protein levels were examined by western blot and immunofluorescence. The results indicated that GANT61 significantly inhibited the proliferation of Daoy cells in a dose-dependent manner, compared with the control group (P<0.05). HE staining revealed that cells had increasingly abnormal protuberance with increasing GANT61 concentration. Flow cytometry analysis also demonstrated that GANT61 induced G1/S arrest and apoptosis of Daoy cells in a dose-dependent manner (P<0.05). Gli1 and CyclinD1 mRNA expression levels were downregulated by GANT61 treatment (P<0.05); similarly, their protein levels were downregulated by GANT61 treatment in a dose-dependent manner (P<0.05). In conclusion, Gli1 expression was significantly associated with CyclinD1 expression in MB. These data demonstrated that Gli1 is an important mediator of the SHH pathway activity in MB, and may be a novel agent for use in combined chemotherapeutic regimens.
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spelling pubmed-52448512017-01-25 Inhibition of the CyclinD1 promoter in response to sonic hedgehog signaling pathway transduction is mediated by Gli1 Lin, Zhongxiao Sheng, Hansong You, Chaoguo Cai, Ming Zhang, Yiping Yu, Li Sheng Yu, Xiaoming Lin, Jian Zhang, Nu Exp Ther Med Articles Medulloblastoma (MB) is the most common malignant tumor of the central nervous system in children. Accumulating evidence suggests a major role for the activation of the sonic hedgehog (SHH) signaling pathway in the development of MB cells; however, the mechanisms underlying the effect of this pathway on tumor survival and growth remain poorly understood. The Gli family zinc finger 1 (Gli1) transcription factor is considered as a mediator of the SHH signaling pathway in MB cells. Therefore, the present study investigated whether the SHH signaling pathway promotes the apoptosis of MB cells via downregulation of Gli1. GANT61, a novel Gli1 inhibitor, is known to have an in vitro activity against tumors. In the current study, Daoy cells were treated with different concentrations of GANT61 for 24 h, and the effect on cell proliferation was assayed by cell counting kit-8 assay. In addition, the cell cycle progression and apoptosis were assayed by flow cytometry analysis and hematoxylin-eosin (HE) staining. The effects of GANT61 treatment on SHH signaling pathway at the mRNA level were assayed by polymerase chain reaction (PCR). To further elucidate the inhibitory effects of GANT61 on the expression of Gli1 and CyclinD1, their protein levels were examined by western blot and immunofluorescence. The results indicated that GANT61 significantly inhibited the proliferation of Daoy cells in a dose-dependent manner, compared with the control group (P<0.05). HE staining revealed that cells had increasingly abnormal protuberance with increasing GANT61 concentration. Flow cytometry analysis also demonstrated that GANT61 induced G1/S arrest and apoptosis of Daoy cells in a dose-dependent manner (P<0.05). Gli1 and CyclinD1 mRNA expression levels were downregulated by GANT61 treatment (P<0.05); similarly, their protein levels were downregulated by GANT61 treatment in a dose-dependent manner (P<0.05). In conclusion, Gli1 expression was significantly associated with CyclinD1 expression in MB. These data demonstrated that Gli1 is an important mediator of the SHH pathway activity in MB, and may be a novel agent for use in combined chemotherapeutic regimens. D.A. Spandidos 2017-01 2016-12-13 /pmc/articles/PMC5244851/ /pubmed/28123507 http://dx.doi.org/10.3892/etm.2016.3969 Text en Copyright: © Lin et al. This is an open access article distributed under the terms of the Creative Commons Attribution-NonCommercial-NoDerivs License (https://creativecommons.org/licenses/by-nc-nd/4.0/) , which permits use and distribution in any medium, provided the original work is properly cited, the use is non-commercial and no modifications or adaptations are made.
spellingShingle Articles
Lin, Zhongxiao
Sheng, Hansong
You, Chaoguo
Cai, Ming
Zhang, Yiping
Yu, Li Sheng
Yu, Xiaoming
Lin, Jian
Zhang, Nu
Inhibition of the CyclinD1 promoter in response to sonic hedgehog signaling pathway transduction is mediated by Gli1
title Inhibition of the CyclinD1 promoter in response to sonic hedgehog signaling pathway transduction is mediated by Gli1
title_full Inhibition of the CyclinD1 promoter in response to sonic hedgehog signaling pathway transduction is mediated by Gli1
title_fullStr Inhibition of the CyclinD1 promoter in response to sonic hedgehog signaling pathway transduction is mediated by Gli1
title_full_unstemmed Inhibition of the CyclinD1 promoter in response to sonic hedgehog signaling pathway transduction is mediated by Gli1
title_short Inhibition of the CyclinD1 promoter in response to sonic hedgehog signaling pathway transduction is mediated by Gli1
title_sort inhibition of the cyclind1 promoter in response to sonic hedgehog signaling pathway transduction is mediated by gli1
topic Articles
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5244851/
https://www.ncbi.nlm.nih.gov/pubmed/28123507
http://dx.doi.org/10.3892/etm.2016.3969
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