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Masume Behruzi1 
, Mehrdad Ghorbanlou1 , Ronak Shabani1 , Auob Rustamzadeh2 , Enam Alhagh Charkhat Gorgich3 , Elham Seidkhani1 , Farnoosh Usefi1 , Mehdi Mehdizadeh1 , Fatemeh Moradi * 4
, Mehrdad Ghorbanlou1 , Ronak Shabani1 , Auob Rustamzadeh2 , Enam Alhagh Charkhat Gorgich3 , Elham Seidkhani1 , Farnoosh Usefi1 , Mehdi Mehdizadeh1 , Fatemeh Moradi * 4
1- Department of Anatomical Sciences, Faculty of Medicine, Iran University of Medical Sciences, Tehran, Iran.
2- Department of Anatomy, School of Medicine, Qazvin University of Medical Sciences, Qazvin, Iran.
3- Department of Anatomy, School of Medicine, Iranshahr University of Medical Sciences, Iranshahr, Iran.
4- Department of Anatomical Sciences, Faculty of Medicine, Iran University of Medical Sciences, Tehran, Iran
2- Department of Anatomy, School of Medicine, Qazvin University of Medical Sciences, Qazvin, Iran.
3- Department of Anatomy, School of Medicine, Iranshahr University of Medical Sciences, Iranshahr, Iran.
4- Department of Anatomical Sciences, Faculty of Medicine, Iran University of Medical Sciences, Tehran, Iran
Abstract:
Introduction: Targeting SMO has been a remarkable strategy in Shh-dependent cancers, especially medulloblastoma. In this manner, GDC-0449 or vismodegib is used as a potent SMO inhibitor with mild toxicity and high affinity. Thus, the main purpose of the present study was to investigate the anti-medulloblastoma efficacy of vismodegib on the DAOY medulloblastoma cell line.
Methods: Human DAOY medulloblastoma cells were cultured in DMEM.50, 80, 100, and 150μM doses of vismodegib were used to treat cells. MTT, Scratch, and trypan blue assay, and also real-time polymerase chain reaction (RT-PCR) and immunofluorescence studies were performed, 24 and 48 hours post-treatment.
Results: MTT and trypan blue assay showed a significant difference in viability, between the control group and treatment groups. Results of the scratch assay showed that in the control group, the cells managed to repair the lesion, while the scratch was disintegrated in higher doses of vismodegib. The expression of SMO, Gli1, and MYCN genes, as the main components of the SHH signaling pathway, had a significant reduction in the vismodegib-treated groups compared to the control group. Also a notable rise in the activation of metastasis-promoting genes (Bax and p53) while a reduction in metastasis-inhibiting gene (Bcl2), was observed.
Conclusion: The results of the current study confirm that vismodegib is a potent inhibitor of the Shh pathway and could be used as a drug in combination with new therapeutic methods, in order to treat medulloblastoma.
Methods: Human DAOY medulloblastoma cells were cultured in DMEM.50, 80, 100, and 150μM doses of vismodegib were used to treat cells. MTT, Scratch, and trypan blue assay, and also real-time polymerase chain reaction (RT-PCR) and immunofluorescence studies were performed, 24 and 48 hours post-treatment.
Results: MTT and trypan blue assay showed a significant difference in viability, between the control group and treatment groups. Results of the scratch assay showed that in the control group, the cells managed to repair the lesion, while the scratch was disintegrated in higher doses of vismodegib. The expression of SMO, Gli1, and MYCN genes, as the main components of the SHH signaling pathway, had a significant reduction in the vismodegib-treated groups compared to the control group. Also a notable rise in the activation of metastasis-promoting genes (Bax and p53) while a reduction in metastasis-inhibiting gene (Bcl2), was observed.
Conclusion: The results of the current study confirm that vismodegib is a potent inhibitor of the Shh pathway and could be used as a drug in combination with new therapeutic methods, in order to treat medulloblastoma.
Type of Study: Original |
Subject:
Cellular and molecular Neuroscience
Received: 2024/03/17 | Accepted: 2024/04/11
Received: 2024/03/17 | Accepted: 2024/04/11
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