Malays. Appl. Biol. (2018) 47(1): 173–179
VALIDATION OF ENDOGENOUS REFERENCE GENES FOR
RELATIVE QUANTITATION STUDIES OF GENE EXPRESSION
IN NASOPHARYNGEAL CARCINOMA
ASHLEY EDWARD ROY SOOSAY1*, ISMAIL MAAMAR MAKHZOUM ALHWIJ1,
PARVATHEE PONNIAH1, ALAN SOO-BENG KHOO2 and CHUN-YIING WONG3
1Faculty of Medicine & Health Sciences, Universiti Malaysia Sarawak, Kota Samarahan, Sarawak
2Institute for Medical Research, Kuala Lumpur
3Sarawak General Hospital, Kuching, Sarawak
*E-mail: This e-mail address is being protected from spambots. You need JavaScript enabled to view it
Accepted 27 February 2018, Published online 31 March 2018
ABSTRACT
Reverse transcription real-time quantitative polymerase chain reaction (RT-qPCR) is a useful molecular contraption in translational biomedical research and clinical settings. RT-qPCR requires normalization. Housekeeping gene (HKG) as reference gene (RG) is commonly used for the relative quantification of the target gene (TG) in gene profiling assays. Normalization requires stably expressed endogenous RG. Recently, RGs were found to be regulated in a various experimental milieu in different tissues. Therefore, it is pertinent to identify HKGs that are stably expressed and are independent of factors influencing the cell. To validate 4 endogenous RGs for the relative quantification of TGs in gene expression analysis performed via RTqPCR in nasopharyngeal carcinoma. The qbase+ software utilizing geNorm analysis identified GAPDH, TBP and YARS as stably expressed HKGs. ACTB was the least stable RG in this study. The most suitable set of RG for NPC gene expression studies include GAPDH, TBP and YARS. No single gene was identified as the best RG for expression study. The RG that can be utilized during RT-qPCR on normal and malignant nasopharyngeal tissue samples is a collection of 3 genes (GAPDH, TBP and YARS) used as an average.
Key words: RT-qPCR, nasopharyngeal carcinoma, normalization, gene expression
