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

Neural Regeneration Research 2022年第6期17卷 1387-1392页

作者： Wei Liu Jie Yu Yi-Fan Wang Qian-Qian Shan Ya-Xian Wang Key Laboratory of Neuroregeneration of Jiangsu and Ministry of Education Co-innovation Center of NeuroregenerationNational Medical Products Administration(NMPA)Key Laboratory for Research and Evaluation of Tissue Engineering Technology ProductsNantong UniversityNantongJiangsu ProvinceChina Department of Nursing The Affiliated Hospital of Nantong UniversityNantongJiangsu ProvinceChina Department of Clinical Medicine Medical School of Nantong UniversityNantongJiangsu ProvinceChina Department of Radiotherapy and Oncology The Affiliated Hospital of Nantong UniversityNantongJiangsu ProvinceChina

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 *** this study,we used rat models of spinal cord hemisection to analyze the expression stability of 13 commonly applied reference genes:Actb,Ankrd27,CypA,Gapdh,Hprt1,Mrpl10,Pgk1,Rictor,Rn18s,Tbp,Ubc,Ubxn11,and *** results show that the expression of Ankrd27,Ubc,and Tbp were stable after spinal cord injury,while Actb was the most unstable internal control ***27,Ubc,Tbp,and Actb were consequently used to investigate the effects of internal control genes with differing stabilities on the normalization of target gene *** 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 *** recommend that Ankrd27,Ubc,and Tbp are used as internal control genes for real-time reverse transcription polymerase chain reaction in spinal cord injury *** study was approved by the Administration Committee of Experimental Animals,Jiangsu Province,China(approval No.20180304-008)on March 4,2018.

关键词： geNorm analysis reference genes internal control genes normalization NormFinder analysis reverse transcription-quantitative polymerase chain reaction spinal cord injury stability of gene expression

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Real-time quantitative PCR array to study drug-induced changes of gene expression in tumor cell lines

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Journal of Cancer Metastasis and Treatment 2017年第1期3卷 90-99页

作者： Stefano Amatori Giuseppe Persico Mirco Fanelli Molecular Pathology Laboratory“PaoLa” Department of Biomolecular SciencesUniversity of Urbino“Carlo Bo”61032 Fano(PU)Italy Department of Experimental Oncology European Institute of Oncology20139 MilanItaly.

Aim:reverse transcription-quantitative polymerase chain reaction(RT-qPCR)is still the“gold standard”for quantitative analysis of mRNA and the study of differentially expressed ***:The authors describe a RT-qPCR array that exploits SYBR Green dye-based detection to perform reliable gene expression analysis on 41 genes involved in several pathways linked to DNA damage response,cell cycle progression,cellular senescence,and programmed cell *** validate the RT-qPCR array,the authors investigated changes of the gene expression profile of HeLa cells treated with two well-characterized antiproliferative molecules such as cisplatin(CDDP)and sodium butyrate(NaBu).Results:The results showed a gene expression profile compatible with both biological and gene expression data already reported in ***:Importantly,the assay allowed the monitoring of additional and not reported gene regulations,indicating that this custom-made RT-qPCR array is a cheap,robust,and rapid tool for the study of drug-induced effects in human biological models.

关键词： reverse transcription-quantitative polymerase chain reaction gene expression cancer treatment

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