title: Identification of Suitable Reference Genes for mRNA Studies in Bone Marrow in a Mouse Model of Hematopoietic Stem Cell Transplantation.
abstract: BACKGROUND
  Bone marrow micro-environment changes during hematopoietic stem cell transplantation (HSCT) with subsequent alteration of genes expression. Quantitative polymerase chain reaction (q-PCR) is a reliable and reproducible technique for the analysis of gene expression. To obtain more accurate results, it is essential to find a reference during HSCT. However, which gene is suitable during HSCT remains unclear. This study aimed to identify suitable reference genes for mRNA studies in bone marrow after HSCT.
  
  
  METHODS
  C57BL/6 mice were treated with either total body irradiation (group T) or busulfan/cyclophosphamide (BU/CY) (group B) followed by infusion of bone marrow cells. Normal mice without treatments were served as a control. All samples (group T + group B + control) were defined as group G. On days 7, 14, and 21 after transplantation, transcription levels of 7 candidate genes, ACTB, B2M, GAPDH, HMBS, HPRT, SDHA, and YWHAZ, in bone marrow cells were measured by use of real-time quantitative PCR. The expression stability of these 7 candidate reference genes were analyzed by 2 statistical software programs, GeNorm and NormFinder.
  
  
  RESULTS
  Our results showed that ACTB displayed the highest expression in group G, with lowest expression of PSDHA in group T and HPRT in groups B and G. Analysis of expression stability by use of GeNorm or NormFinder demonstrated that expression of B2M in bone marrow were much more stable during HSCT, compared with other candidate genes including commonly used reference genes GAPDH and ACTB.
  
  
  CONCLUSIONS
  ACTB could be used as a suitable reference gene for mRNA studies in bone marrow after HSCT.
authors: Department of Clinical Laboratory, the Affiliated Hospital of Xuzhou Medical College, Xuzhou, China.
authors: H
authors: H
authors: Li
authors: Blood Diseases Institute, Xuzhou Medical College, Xuzhou, China; Key Laboratory of Bone Marrow Stem Cell, Jiangsu Province, Xuzhou, China.
authors: C
authors: C
authors: Chen
authors: Blood Diseases Institute, Xuzhou Medical College, Xuzhou, China; Key Laboratory of Bone Marrow Stem Cell, Jiangsu Province, Xuzhou, China.
authors: H
authors: H
authors: Yao
authors: Blood Diseases Institute, Xuzhou Medical College, Xuzhou, China; Key Laboratory of Bone Marrow Stem Cell, Jiangsu Province, Xuzhou, China.
authors: X
authors: X
authors: Li
authors: Blood Diseases Institute, Xuzhou Medical College, Xuzhou, China; Key Laboratory of Bone Marrow Stem Cell, Jiangsu Province, Xuzhou, China.
authors: N
authors: N
authors: Yang
authors: Blood Diseases Institute, Xuzhou Medical College, Xuzhou, China; Key Laboratory of Bone Marrow Stem Cell, Jiangsu Province, Xuzhou, China; Department of Hematology, the Affiliated Hospital of Xuzhou Medical College, Xuzhou, China.
authors: J
authors: J
authors: Qiao
authors: Blood Diseases Institute, Xuzhou Medical College, Xuzhou, China; Key Laboratory of Bone Marrow Stem Cell, Jiangsu Province, Xuzhou, China; Department of Hematology, the Affiliated Hospital of Xuzhou Medical College, Xuzhou, China. Electronic address: lihmd@163.com.
authors: K
authors: K
authors: Xu
authors: Blood Diseases Institute, Xuzhou Medical College, Xuzhou, China; Key Laboratory of Bone Marrow Stem Cell, Jiangsu Province, Xuzhou, China; Department of Hematology, the Affiliated Hospital of Xuzhou Medical College, Xuzhou, China. Electronic address: zengly2000@163.com.
authors: L
authors: L
authors: Zeng
mesh_terms: D000818:Animals; D001853:Bone Marrow / Q000378:metabolism; D001854:Bone Marrow Cells / Q000502:physiology*; D004195:Disease Models, Animal; D015870:Gene Expression / Q000235:genetics; D020869:Gene Expression Profiling; D005796:Genes / Q000235:genetics*; D018380:Hematopoietic Stem Cell Transplantation; D051379:Mice; D008807:Mice, Inbred BALB C; D008810:Mice, Inbred C57BL; D012333:RNA, Messenger / Q000235:genetics*; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods; D012984:Software
chemical_list: D012333:RNA, Messenger