Validation of Reference Genes for Reliable Gene Expression Study in Human Glioblastoma Multiforme and 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) allows selective 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) using a validated internal control across GBM and NSC. This study is the first evaluation on the suitability of nine reference genes as the internal control in human glioblastoma cell line (DBTRG-05MG) and H9-derived human NSC (H9-hNSC) using RT-qPCR. The results were analyzed using geNorm, NormFinder and BestKeeper algorithms. The analyses revealed PPIA as the most stably expressed gene across DBTRG-05MG and H9-hNSC. Both geNorm and NormFinder 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. Keywords: Glioblastoma multiforme; Neural stem cell; Reverse transcription quantitative real-time polymerase chain reaction; reference genes
Authors and Affiliations
Kang In Nee
Keywords
Related Articles
- EP ID EP418347
- DOI -
- Views 117
- Downloads 0
Strategic Approaches of Controlling Variables in RP-HPLC Method Development and Validation: A Review
High Performance Liquid chromatography is the principal technique in accessing the drug, its various impurities and drug related degradants that can form on synthesis or storage in pharmaceutical products. It results in...
DRUG USE PATTERN IN MANAGEMENT OF RESPIRATORY EMERGENCIES IN MICU-A PROSPECTIVE OBSERVATIONAL STUDY
BACKGROUND: Drug utilization studies are useful tool to facilitate rational prescribing of drugs. The objective of the study is to analyze drug utilization in respiratory emergencies using WHO Prescribing Indicators, in...
Effect of quercetin on estrogen receptor α and β expression in uterine tissues in ovariectomized rats
Objective To explore the effect of quercetin on mRNA expressions of estrogen receptor α (ERα) and estrogen receptor β (ERβ) in ovariectomized (OVX) Sprague-Dawley (SD) rats. Method Thirty-two SD rats were randomly divide...
ISOLATION AND CHARACTERIZATION OF A FLAVONOID FROM ETHANOLIC EXTRACT OF ALTERNANTHERA SESSILIS LINN.
This study was performed to isolate and characterize the flavonoid present in Alternanthera sessilis Linn. Direct soxhlet extraction process was adopted for extraction by using 95% ethanol and vacuum evaporator was used...
OCCLUSION THERAPY TREATMENT IN AMBLYOPIC EYE
Amblyopia is the most common visual disorder of childhood, yet the contributions of the two principal treatments (spectacle wear and occlusion) to outcome are unknown. The MOTAS study investigated the dose-response relat...