中国神经再生研究(英文版)

中国神经再生研究(英文版) ›› 2022, Vol. 17 ›› Issue (on line): 1387-1392.

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Selection of suitable internal controls for gene expression normalization in rats with spinal cord injury

  

  • 出版日期:2022-01-01 发布日期:2021-10-08

Wei Liu1, #, Jie Yu2, 3, #, Yi-Fan Wang3, Qian-Qian Shan4, *, Ya-Xian Wang1, *   

  1. 1Key Laboratory of Neuroregeneration of Jiangsu and Ministry of Education, Co-innovation Center of Neuroregeneration, National Medical Products Administration (NMPA) Key Laboratory for Research and Evaluation of Tissue Engineering Technology Products, Nantong University, Nantong, Jiangsu Province, China; 2Department of Nursing, The Affiliated Hospital of Nantong University, Nantong, Jiangsu Province, China; 3Department of Clinical Medicine, Medical School of Nantong University, Nantong, Jiangsu Province, China; 4Department of Radiotherapy and Oncology, The Affiliated Hospital of Nantong University, Nantong, Jiangsu Province, China
  • Online:2022-01-01 Published:2021-10-08
  • Contact: Qian-Qian Shan, shanqianqian1986@126.com; Ya-Xian Wang, MD, wyx1984@ntu.edu.cn.
  • Supported by:
    This study was supported by the National Natural Science Foundation of China, No. 81901257 (to YXW); the Natural Science Foundation of Jiangsu Province of China, No. BK20180951 (to YXW); Postgraduate Research and Practice Innovation Program of Jiangsu Province of China, No. KYCX20_2818 (to WL); and Priority Academic Program Development of Jiangsu Higher Education Institutions (PAPD, to Key Laboratory of Neuroregeneration of Jiangsu and Ministry of Education).

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摘要： https://orcid.org/0000-0001-5854-9352 (Ya-Xian Wang); https://orcid.org/0000-0002-9742-8139 (Wei Liu); 
https://orcid.org/0000-0002-6271-4346 (Qian-Qian Shan)

Abstract: There is a lack of systematic research on the expression of internal control genes used for gene expression normalization in real-time reverse transcription polymerase chain reaction in spinal cord injury research. In this study, we used rat models of spinal cord hemisection to analyze the expression stability of 13 commonly applied housekeeping genes: Actb, Ankrd27, CypA, Gapdh, Hprt1, Mrpl10, Pgk1, Rictor, Rn18s, Tbp, Ubc, Ubxn11, and Ywhaz. Our results show that the expression of Ankrd27, Ubc, and Tbp were stable after spinal cord injury, while Actb was the most unstable internal control gene. Ankrd27, Ubc, Tbp, and Actb were consequently used to investigate the effects of internal control genes with differing stabilities on the normalization of target gene expression. Target gene expression levels and changes over time were similar when Ankrd27, Ubc, and Tbp were used as internal controls but different when Actb was used as an internal control. We recommend that Ankrd27, Ubc, and Tbp are used as internal control genes for real-time reverse transcription polymerase chain reaction in spinal cord injury research. This study was approved by the Administration Committee of Experimental Animals, Jiangsu Province, China (approval No. 20180304-008) on March 4, 2018.

Key words: geNorm analysis, housekeeping genes, internal control genes, normalization, NormFinder analysis, reverse transcription-quantitative polymerase chain reaction,  , spinal cord injury, stability of gene expression