The anterior gradient homologue 2 (AGR2) co-localises with the glucose-regulated protein 78 (GRP78) in cancer stem cells, and is critical for the survival and drug resistance of recurrent glioblastoma: in situ and in vitro analyses

Cancer Cell International

Research

Hans-Juergen Schulten¹  Ahmed M. Al-Hejin²  Adeel Chaudhary³  Mohammed Bangash⁴  Saleh Baeesa⁴  Abdulla Ahmed A. Salwati⁵  Rinad Algehani⁵  Alazouf Alhowity⁵  Reem Alsereihi⁶  Deema Hussein⁷  Torki Al Zughaibi⁷  Mohamad Saka⁷  Adel Abuzenadah⁸  Fahad Alghamdi⁹  Richard Cross^1,10 

[1] Center of Excellence in Genomic Medicine Research, Department of Medical Laboratory Technology, Faculty of Applied Medical Sciences, King Abdulaziz University, 21589, Jeddah, Saudi Arabia;Department of Biological Sciences, Faculty of Science, King Abdulaziz University, 80203, 21589, Jeddah, Saudi Arabia;Department of Medical Laboratory Sciences, Faculty of Applied Medical Sciences, King Abdulaziz University, 21589, Jeddah, Saudi Arabia;Centre of Innovation for Personalized Medicine, Department of Medical Laboratory Technology, Faculty of Applied Medical Sciences, King Abdulaziz University, 21589, Jeddah, Saudi Arabia;Division of Neurosurgery, Faculty of Medicine, King Abdulaziz University, 21589, Jeddah, Saudi Arabia;King Fahd Medical Research Center, King Abdulaziz University, 80216, 21589, Jeddah, Saudi Arabia;King Fahd Medical Research Center, King Abdulaziz University, 80216, 21589, Jeddah, Saudi Arabia;Department of Biological Sciences, Faculty of Science, King Abdulaziz University, 80203, 21589, Jeddah, Saudi Arabia;College of Health Sciences, Al-Rayan Colleges, 41411, Madinah AL-Munawarah, Saudi Arabia;King Fahd Medical Research Center, King Abdulaziz University, 80216, 21589, Jeddah, Saudi Arabia;Department of Medical Laboratory Sciences, Faculty of Applied Medical Sciences, King Abdulaziz University, 21589, Jeddah, Saudi Arabia;King Fahd Medical Research Center, King Abdulaziz University, 80216, 21589, Jeddah, Saudi Arabia;Department of Medical Laboratory Sciences, Faculty of Applied Medical Sciences, King Abdulaziz University, 21589, Jeddah, Saudi Arabia;Center of Excellence in Genomic Medicine Research, Department of Medical Laboratory Technology, Faculty of Applied Medical Sciences, King Abdulaziz University, 21589, Jeddah, Saudi Arabia;Centre of Innovation for Personalized Medicine, Department of Medical Laboratory Technology, Faculty of Applied Medical Sciences, King Abdulaziz University, 21589, Jeddah, Saudi Arabia;Pathology Department, Faculty of Medicine, King Abdulaziz University, 21589, Jeddah, Saudi Arabia;School of Engineering and Sustainable Development, Emerging Technologies Research Centre (EMTERC), De Montfort University, The Gateway, LE1 9BH, Leicester, UK;

关键词: Glioblastoma;    Cancer stem cells (CSCs);    Anterior gradient homologue 2 (AGR2);    Glucose-regulated protein 78 (GRP78);    Drug resistance;   

DOI  :  10.1186/s12935-022-02814-5

 received in 2022-09-12, accepted in 2022-11-28,  发布年份 2022

来源: Springer

【 摘 要 】

BackgroundGlioblastomas (GBs) are characterised as one of the most aggressive primary central nervous system tumours (CNSTs). Single-cell sequencing analysis identified the presence of a highly heterogeneous population of cancer stem cells (CSCs). The proteins anterior gradient homologue 2 (AGR2) and glucose-regulated protein 78 (GRP78) are known to play critical roles in regulating unfolded protein response (UPR) machinery. The UPR machinery influences cell survival, migration, invasion and drug resistance. Hence, we investigated the role of AGR2 in drug-resistant recurrent glioblastoma cells.MethodsImmunofluorescence, biological assessments and whole exome sequencing analyses were completed under in situ and in vitro conditions. Cells were treated with CNSTs clinical/preclinical drugs taxol, cisplatin, irinotecan, MCK8866, etoposide, and temozolomide, then resistant cells were analysed for the expression of AGR2. AGR2 was repressed using single and double siRNA transfections and combined with either temozolomide or irinotecan.ResultsGenomic and biological characterisations of the AGR2-expressed Jed66_GB and Jed41_GB recurrent glioblastoma tissues and cell lines showed features consistent with glioblastoma. Immunofluorescence data indicated that AGR2 co-localised with the UPR marker GRP78 in both the tissue and their corresponding primary cell lines. AGR2 and GRP78 were highly expressed in glioblastoma CSCs. Following treatment with the aforementioned drugs, all drug-surviving cells showed high expression of AGR2. Prolonged siRNA repression of a particular region in AGR2 exon 2 reduced AGR2 protein expression and led to lower cell densities in both cell lines. Co-treatments using AGR2 exon 2B siRNA in conjunction with temozolomide or irinotecan had partially synergistic effects. The slight reduction of AGR2 expression increased nuclear Caspase-3 activation in both cell lines and caused multinucleation in the Jed66_GB cell line.ConclusionsAGR2 is highly expressed in UPR-active CSCs and drug-resistant GB cells, and its repression leads to apoptosis, via multiple pathways.

【 授权许可】

CC BY   
© The Author(s) 2022

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