title: BCL11B suppression does not influence CD34(+) cell differentiation and proliferation.
abstract: The B-cell chronic lymphocytic leukemia (CLL)/lymphoma 11B (BCL11B) gene plays a critical role in T-cell differentiation and proliferation. However, little is understood about the role of BCL11B in human hematopoietic stem/progenitor cells. Small interfering RNA (siRNA)-mediated  suppression of the BCL11B was shown to induce apoptosis in human T-cell acute lymphoblastic leukemia cells. To further characterize the role of BCL11B in hematopoietic stem/progenitor cells and assess the safety of siRNA-mediated targeted therapy, the in vitro differentiation  and proliferation of CD34(+) cells after BCL11B-siRNA935 treatment were studied. CD34(+) cells were sorted from three cases of umbilical cord blood by the magnetic activated cell sorting technique, and the purity was identified by flow cytometry. BCL11B-siRNA935  was delivered into CD34(+) cells by nucleofection and the BCL11B expression level was analyzed by quantitative real-time polymerase chain reaction. Erythroid burst-forming units (BFU-E), granulocyte/macrophage colony-forming units (CFU-GM), and megakaryocyte colony-forming  units (CFU-Meg) were assessed using BCL11B-siRNA935-treated CD34(+) cells by the methylcellulose semi-solid culture method. The BCL11B expression level in CD34(+) cells was significantly lower than that in Molt-4 cells and peripheral blood mononuclear cells  from healthy individuals. An approximate one-fold reduction in the BCL11B mRNA level was observed 24 hours post-transfection with BCL11B-siRNA935. However, there was no significant difference on the colony formation ability of BFU-E, CFU-GM, and CFU-Meg for CD34(+) cells  between the BCL11B-siRNA935-treated and mock-transfected groups (P > 0.05). BCL11B suppression by RNA interference had no significant influence on the differentiation and proliferation of CD34(+) cells. In conclusion, the BCL11B-siRNA935 used  in this study may be safe, and BCL11B may be considered a new candidate for targeted gene therapy in T-cell malignancies.
authors.affiliation: Institute of Hematology, Medical College, Jinan University, Guangzhou, PR China.
authors.forename: Qi
authors.initials: Q
authors.lastname: Shen
authors.forename: Xin
authors.initials: X
authors.lastname: Huang
authors.forename: Si
authors.initials: S
authors.lastname: Chen
authors.forename: Lijian
authors.initials: L
authors.lastname: Yang
authors.forename: Shaohua
authors.initials: S
authors.lastname: Chen
authors.forename: Bo
authors.initials: B
authors.lastname: Li
authors.forename: Xiuli
authors.initials: X
authors.lastname: Wu
authors.forename: Piotr
authors.initials: P
authors.lastname: Grabarczyk
authors.forename: Grzegorz K
authors.initials: GK
authors.lastname: Przybylski
authors.forename: Christian A
authors.initials: CA
authors.lastname: Schmidt
authors.forename: Yangqiu
authors.initials: Y
authors.lastname: Li
mesh_terms: D000328:Adult; D018952:Antigens, CD34 / Q000378:metabolism*; D002454:Cell Differentiation / Q000235:genetics*; D002460:Cell Line; D049109:Cell Proliferation; D005260:Female; D015870:Gene Expression; D006412:Hematopoietic Stem Cells / Q000166:cytology* / Q000378:metabolism*; D006801:Humans; D008297:Male; D008875:Middle Aged; D011247:Pregnancy; D034622:RNA Interference; D012097:Repressor Proteins / Q000235:genetics* / Q000378:metabolism; D014162:Transfection; D025521:Tumor Suppressor Proteins / Q000235:genetics* / Q000378:metabolism; D055815:Young Adult
chemical_list: D018952:Antigens, CD34; C479740:BCL11B protein, human; D012097:Repressor Proteins; D025521:Tumor Suppressor Proteins