Validation of Reference Genes for Reliable Gene Expression Study in Human Glioblastoma Multiformeand Neural Stem Cell Lines
Journal Title: Journal of Biomedical and Pharmaceutical Research - Year 2016, Vol 5, Issue 5
Abstract
Glioblastoma multiforme (GBM) is the most deadly malignant brain tumor in adults. GBM has a histologically heterogeneous group of neoplastic cells which are postulated to be the mediator of anti-cancer therapy resistance and tumor relapse. Therefore, it is important to design precise therapies targeting the GBM. The tumor homing of human neural stem cells (NSC) allowsselective drug delivery to kill GBM. The therapeutic effects of NSC for GBM can be assessed by reverse transcription real-time polymerase chain reaction (RT-qPCR) usinga validated internal control across GBM and NSC. This study is the first evaluation onthe suitability of nine reference genes as the internal control in human glioblastoma cell line (DBTRG-05MG) andH9-derived human NSC (H9-hNSC) using RT-qPCR. The results were analyzed using geNorm, Norm Finder and Best keeper algorithms. The analyses revealed PPIA as the most stably expressed gene across DBTRG-05MG and H9-hNSC.Both ge Norm and Norm Finder suggested PPIA and RPL13A as the best combination of reference genes for RT-qPCR normalization across DBTRG-05MG and H9-hNSC. HPRT1 was ranked as the least stably expressed gene between DBTRG-05MG and H9-hNSC by all three analyses. In conclusion, PPIA and RPL13A were proposed as the best reference genes for these two cell types.
Authors and Affiliations
Kang In Nee| School of Health Sciences, Health Campus, UniversitiSains Malaysia, 16150 KubangKerian, Kelantan, Malaysia., Abdul Rahman Izaini Ghani| School of Medical Sciences, Health Campus, UniversitiSains Malaysia, 16150 KubangKerian, Kelantan, Malaysia., Shaharum Shamsuddin| School of Health Sciences, Health Campus, UniversitiSains Malaysia, 16150 KubangKerian, Kelantan, Malaysia., Tan Suat Cheng| School of Health Sciences, Health Campus, UniversitiSains Malaysia, 16150 KubangKerian, Kelantan, Malaysia.
Keywords
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