Identification of valid reference genes for mRNA and microRNA normalisation in prostate cancer cell lines

doi:10.1038/s41598-018-19458-z

CORC > 昆明动物研究所 > 昆明动物研究所 > 遗传资源与进化国家重点实验室 > 分子进化基因组学

	Identification of valid reference genes for mRNA and microRNA normalisation in prostate cancer cell lines
	Lin-Lin Liu 3,5; Jie Lin 5; Teng-Fei Ma 3,5; Wei-Jie Sun 3,5; Newton O. Otecko 1,4; Ya-Ping Zhang 1,3; Hui Zhao 2,3; Li-Xia Guo 3,5; Si-Qi Wang 3,5
刊名	SCIENTIFIC REPORTS
	2018
卷号	8 期号:1 页码:1949
DOI	10.1038/s41598-018-19458-z
英文摘要	RT-qPCR offers high sensitivity, for accurate interpretations of qPCR results however, normalisationusing suitable reference genes is fundamental. Androgens can regulate transcriptional expressionincluding reference gene expression in prostate cancer. In this study, we evaluated ten mRNA and sixnon-protein coding RNA reference genes in five prostate cell lines under varied dihydrotestosterone(DHT) treatments. We validated the effects of DHT-treatments using media containing charcoal-stripped serum prior to DHT stimulation on the test samples by Western blot experiments. Referencegene expression stability was analysed using three programs (geNorm, NormFinder and BestKeeper),and the recommended comprehensive ranking is provided. Our results reveal that ACTB and GAPDH,and miR-16 and miR-1228-3p are the most suitable mRNA and miRNA reference genes across all celllines, respectively. Considering prostate cancer cell types, ACTB/GAPDH and ACTB/HPRT1 are the mostsuitable reference gene combinations for mRNA analysis, and miR-16/miR-1228-3p and RNU6-2/RNU43for miRNA analysis in AR+, and AR− and normal cell lines, respectively. Comparison of relative targetgene (PCA3 and miR-141) expression reveals different patterns depending on reference genes used fornormalisation. To our knowledge, this is the first report on validation of reference genes under differentDHT treatments in prostate cancer cells. This study provides insights for discovery of reliable DHT-regulated genes in prostate cells.
语种	英语
内容类型	期刊论文
源URL	[http://159.226.149.26:8080/handle/152453/12396]
专题	昆明动物研究所_分子进化基因组学
通讯作者	Ya-Ping Zhang
作者单位	1.State Key Laboratory of Genetic Resources and Evolution & Yunnan Laboratory of Molecular Biology of Domestic Animals & Germplasm Bank of Wild Species, Kunming Institute of Zoology, Chinese Academy of Sciences, Kunming, 650223, China. 2.Key Laboratory for Animal Genetic Diversity and Evolution of High Education in Yunnan Province, Yunnan University, Kunming, Yunnan, China. 3.State Key Laboratory for Conservation and Utilization of Bio-resource, Yunnan University, Kunming, China. 4.Kunming College of Life Science, University of Chinese Academy of Sciences, Kunming, 650204, China. Hui Zhao, Teng-Fei Ma and Jie Lin contributed equally to this work. Correspondence and requests for materials should be addressed to Y.-P.Z. 5.School of Life Science, Yunnan University, Kunming, 650091, China.
推荐引用方式 GB/T 7714	Lin-Lin Liu,Jie Lin,Teng-Fei Ma,et al. Identification of valid reference genes for mRNA and microRNA normalisation in prostate cancer cell lines[J]. SCIENTIFIC REPORTS,2018,8(1):1949.
APA	Lin-Lin Liu.,Jie Lin.,Teng-Fei Ma.,Wei-Jie Sun.,Newton O. Otecko.,...&Si-Qi Wang.(2018).Identification of valid reference genes for mRNA and microRNA normalisation in prostate cancer cell lines.SCIENTIFIC REPORTS,8(1),1949.
MLA	Lin-Lin Liu,et al."Identification of valid reference genes for mRNA and microRNA normalisation in prostate cancer cell lines".SCIENTIFIC REPORTS 8.1(2018):1949.