Identification of valid reference genes for gene expression studies of human stomach cancer by reverse transcription-qPCR
作者： Hyun-Wook Rho , Byoung-Chan Lee , Eun-Seok Choi , Il-Ju Choi , Yeon-Su Lee
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摘要: Reverse transcription quantitative real-time polymerase chain reaction (RT-qPCR) is a powerful method for the analysis of gene expression. Target expression levels are usually normalized to consistently expressed reference also known as internal standard, in same sample. However, much effort has not been expended thus far search genes suitable study stomach cancer using RT-qPCR, although selection optimal critical interpretation results. We assessed suitability six possible genes, beta-actin (ACTB), glyceraldehydes-3-phosphate dehydrogenase (GAPDH), hypoxanthine phosphoribosyl transferase 1 (HPRT1), beta-2-microglobulin (B2M), ribosomal subunit L29 (RPL29) and 18S RNA (18S rRNA) 20 normal tumor tissue pairs patients 6 cell lines, by RT-qPCR. Employing stability analyses NormFinder geNorm algorithms we determined order performance these their variation values. This RT-qPCR showed that there statistically significant (p < 0.05) differences HPRT1 rRNA 'normal-' versus 'tumor tissues'. The suggest B2M-GAPDH, best combination 'stomach lines'; RPL29-HPRT1, 'all tissues'; ACTB-18S rRNA, lines identified B2M lines', RPL29-B2M tissues', rRNA-ACTB comparisons target gene, GPNMB, different depend on single or combination. validated RPL29 combination, B2M-GAPDH lines'. Use should provide more exact differential expressions at level cancer.
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