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Gene expression profiling and in vitro functional studies reveal RAD54L as a potential therapeutic target in multiple myeloma

Background: Current advances in the molecular biology of multiple myeloma (MM) are not sufficient to fully delineate the genesis and development of this disease. Objective: This study aimed to identify molecular targets underlying MM pathogenesis. Methods: mRNA expression profiling for 29 samples (1...

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Main Authors: Bong, Ivyna Pau Ni, Ng, Ching Ching, Othman, Norodiyah, Esa, Ezalia
Format: Article
Published: Springer Verlag (Germany) 2022
Subjects:
QH301 Biology
Online Access:http://eprints.um.edu.my/43624/
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spelling my.um.eprints.436242023-10-23T03:50:37Z http://eprints.um.edu.my/43624/ Gene expression profiling and in vitro functional studies reveal RAD54L as a potential therapeutic target in multiple myeloma Bong, Ivyna Pau Ni Ng, Ching Ching Othman, Norodiyah Esa, Ezalia QH301 Biology Background: Current advances in the molecular biology of multiple myeloma (MM) are not sufficient to fully delineate the genesis and development of this disease. Objective: This study aimed to identify molecular targets underlying MM pathogenesis. Methods: mRNA expression profiling for 29 samples (19 MM samples, 7 MM cell lines and 3 controls) were obtained using microarray. We evaluated the in vitro effects of RAD54L gene silencing on the proliferation, apoptosis and cell cycle distribution in KMS-28BM human MM cells using siRNA approach. Cell proliferation was determined by MTS assay while apoptosis and cell cycle distribution were analysed with flow cytometry. Gene and protein expression was evaluated using RT-qPCR and ELISA, respectively. Results: Microarray results revealed a total of 5124 differentially expressed genes (DEGs), in which 2696 and 2428 genes were up-regulated and down-regulated in MM compared to the normal controls, respectively (fold change ≥ 2.0; P < 0.05). Up-regulated genes (RAD54L, DIAPH3, SHCBP1, SKA3 and ANLN) and down-regulated genes (HKDC1, RASGRF2, CYSLTR2) have never been reported in association with MM. Up-regulation of RAD54L was further verified by RT-qPCR (P < 0.001). In vitro functional studies revealed that RAD54L gene silencing significantly induced growth inhibition, apoptosis (small changes) and cell cycle arrest in G0/G1 phase in KMS-28BM (P < 0.05). Silencing of RAD54L also decreased its protein level (P < 0.05). Conclusions: This study has identified possible molecular targets underlying the pathogenesis of MM. For the first time, we reveal RAD54L as a potential therapeutic target in MM, possibly functioning in the cell cycle and checkpoint control. © 2022, The Author(s). Springer Verlag (Germany) 2022-06 Article PeerReviewed Bong, Ivyna Pau Ni and Ng, Ching Ching and Othman, Norodiyah and Esa, Ezalia (2022) Gene expression profiling and in vitro functional studies reveal RAD54L as a potential therapeutic target in multiple myeloma. Genes & Genomics, 44 (8). 957 -966. ISSN 1976-9571, DOI https://doi.org/10.1007/s13258-022-01272-7 <https://doi.org/10.1007/s13258-022-01272-7>. 10.1007/s13258-022-01272-7
institution Universiti Malaya
building UM Library
collection Institutional Repository
continent Asia
country Malaysia
content_provider Universiti Malaya
content_source UM Research Repository
url_provider http://eprints.um.edu.my/
topic QH301 Biology
spellingShingle QH301 Biology
Bong, Ivyna Pau Ni
Ng, Ching Ching
Othman, Norodiyah
Esa, Ezalia
Gene expression profiling and in vitro functional studies reveal RAD54L as a potential therapeutic target in multiple myeloma
description Background: Current advances in the molecular biology of multiple myeloma (MM) are not sufficient to fully delineate the genesis and development of this disease. Objective: This study aimed to identify molecular targets underlying MM pathogenesis. Methods: mRNA expression profiling for 29 samples (19 MM samples, 7 MM cell lines and 3 controls) were obtained using microarray. We evaluated the in vitro effects of RAD54L gene silencing on the proliferation, apoptosis and cell cycle distribution in KMS-28BM human MM cells using siRNA approach. Cell proliferation was determined by MTS assay while apoptosis and cell cycle distribution were analysed with flow cytometry. Gene and protein expression was evaluated using RT-qPCR and ELISA, respectively. Results: Microarray results revealed a total of 5124 differentially expressed genes (DEGs), in which 2696 and 2428 genes were up-regulated and down-regulated in MM compared to the normal controls, respectively (fold change ≥ 2.0; P < 0.05). Up-regulated genes (RAD54L, DIAPH3, SHCBP1, SKA3 and ANLN) and down-regulated genes (HKDC1, RASGRF2, CYSLTR2) have never been reported in association with MM. Up-regulation of RAD54L was further verified by RT-qPCR (P < 0.001). In vitro functional studies revealed that RAD54L gene silencing significantly induced growth inhibition, apoptosis (small changes) and cell cycle arrest in G0/G1 phase in KMS-28BM (P < 0.05). Silencing of RAD54L also decreased its protein level (P < 0.05). Conclusions: This study has identified possible molecular targets underlying the pathogenesis of MM. For the first time, we reveal RAD54L as a potential therapeutic target in MM, possibly functioning in the cell cycle and checkpoint control. © 2022, The Author(s).
format Article
author Bong, Ivyna Pau Ni
Ng, Ching Ching
Othman, Norodiyah
Esa, Ezalia
author_facet Bong, Ivyna Pau Ni
Ng, Ching Ching
Othman, Norodiyah
Esa, Ezalia
author_sort Bong, Ivyna Pau Ni
title Gene expression profiling and in vitro functional studies reveal RAD54L as a potential therapeutic target in multiple myeloma
title_short Gene expression profiling and in vitro functional studies reveal RAD54L as a potential therapeutic target in multiple myeloma
title_full Gene expression profiling and in vitro functional studies reveal RAD54L as a potential therapeutic target in multiple myeloma
title_fullStr Gene expression profiling and in vitro functional studies reveal RAD54L as a potential therapeutic target in multiple myeloma
title_full_unstemmed Gene expression profiling and in vitro functional studies reveal RAD54L as a potential therapeutic target in multiple myeloma
title_sort gene expression profiling and in vitro functional studies reveal rad54l as a potential therapeutic target in multiple myeloma
publisher Springer Verlag (Germany)
publishDate 2022
url http://eprints.um.edu.my/43624/
_version_ 1781704685137166336
score 13.211869

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