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

doi:10.1038/s41598-018-19458-z

	Identification of valid reference genes for mRNA and microRNA normalisation in prostate cancer cell lines
	Hui Zhao 1,2; Teng-Fei Ma 1,3; Jie Lin 3; Lin-Lin Liu 1,3; Wei-Jie Sun 1,3; Li-Xia Guo 1,3; Si-Qi Wang 1,3; Newton O. Otecko 4,5; Ya-Ping Zhang1,4
	2018
发表期刊	SCIENTIFIC REPORTS
卷号	8 期号:1 页码:1949
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.
DOI	10.1038/s41598-018-19458-z
语种	英语
引用统计	被引频次：22[WOS] [WOS记录] [WOS相关记录]
文献类型	期刊论文
条目标识符	http://ir.kiz.ac.cn/handle/152453/12396
专题	科研部门_分子进化与基因组多样性（张亚平）
通讯作者	Ya-Ping Zhang
作者单位	1.State Key Laboratory for Conservation and Utilization of Bio-resource, Yunnan University, Kunming, China. 2.Key Laboratory for Animal Genetic Diversity and Evolution of High Education in Yunnan Province, Yunnan University, Kunming, Yunnan, China. 3.School of Life Science, Yunnan University, Kunming, 650091, China. 4.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. 5.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.
推荐引用方式 GB/T 7714	Hui Zhao,Teng-Fei Ma,Jie Lin,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	Hui Zhao.,Teng-Fei Ma.,Jie Lin.,Lin-Lin Liu.,Wei-Jie Sun.,...&Ya-Ping Zhang.(2018).Identification of valid reference genes for mRNA and microRNA normalisation in prostate cancer cell lines.SCIENTIFIC REPORTS,8(1),1949.
MLA	Hui Zhao,et al."Identification of valid reference genes for mRNA and microRNA normalisation in prostate cancer cell lines".SCIENTIFIC REPORTS 8.1(2018):1949.

条目包含的文件
文件名称/大小	文献类型	版本类型	开放类型	使用许可
212.pdf（1814KB）	期刊论文	出版稿	开放获取	CC BY-NC-SA	请求全文