Preferential expression of cytochrome CYP CYP2R1 but not CYP1B1 in human cord blood hematopoietic stem and progenitor cells.
10.1016/j.apsb.2014.10.003
- Author:
Shuoqi XU
1
;
Zhihua REN
2
;
Yanan WANG
3
;
Xinxin DING
1
;
Yongping JIANG
4
;
Author Information
1. Biopharmaceutical R&D Center, Chinese Academy of Medical Sciences & Peking Union Medical College, Suzhou 215126, China.
2. Biopharmagen Corp., Suzhou 215126, China.
3. Department of Clinical Laboratory, Suzhou Municipal Affiliated Hospital of Nanjing Medical University, Suzhou 215002, China.
4. Biopharmaceutical R&D Center, Chinese Academy of Medical Sciences & Peking Union Medical College, Suzhou 215126, China
- Publication Type:Journal Article
- Keywords:
CBHSPCs, cord blood HSPCs;
CD34+;
CYP, cytochrome P450;
CYP1B1;
CYP2R1;
Ct, threshold cycle;
Cytochrome P450;
FACS, fluorescence-activated cell sorting;
Gene expression;
HSPCs, hematopoietic stem and early progenitor cells;
Hematopoietic stem cells;
MNCs, mononuclear cells;
OD, optical density;
PCR, polymerase chain reaction;
PE, R-phycoerythrin;
RT, reverse transcription;
bp, base pair;
kbp, kilobase pair
- From:
Acta Pharmaceutica Sinica B
2014;4(6):464-469
- CountryChina
- Language:English
-
Abstract:
Cytochrome P450 (CYP) enzymes metabolize numerous endogenous substrates, such as retinoids, androgens, estrogens and vitamin D, that can modulate important cellular processes, including proliferation, differentiation and apoptosis. The aim of this study is to characterize the expression of CYP genes in CD34+ human cord blood hematopoietic stem and early progenitor cells (CBHSPCs) as a first step toward assessment of the potential biological functions of CYP enzymes in regulating the expansion or differentiation of these cells. CD34+ CBHSPCs were purified from umbilical cord blood via antibody affinity chromatography. Purity of CD34+ CBHSPCs was assessed using fluorescence-activated cell sorting. RNA was isolated from purified CD34+ CBHSPCs and total mononuclear cells (MNCs) for RNA-PCR analysis of CYP expression. Fourteen human CYPs were detected in the initial screening with qualitative RT-PCR in CD34+ CBHSPCs. Further quantitative RNA-PCR analysis of the detected CYP transcripts yielded evidence for preferential expression of CYP2R1 in CD34+ CBHSPCs relative to MNCs; and for greater expression of CYP1B1 in MNCs relative to CD34+ CBHSPCs. These findings provide the basis for further studies on possible functions of CYP2R1 and CYP1B1 in CBHSPCs׳ proliferation and/or differentiation and their potential utility as targets for drugs designed to modulate CD34+ CBHSPC expansion or differentiation.
