Membranebound stem cell factor increases proliferation and colonyformation of leukemia cell line K562

doi:10.3872/j.issn.1007-385X.2009.1.003

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Archive > Volume 16 Issue 1 > 2009,16(1):6-11. DOI:10.3872/j.issn.1007-385X.2009.1.003 Prev Next

Basic Research

Membranebound stem cell factor increases proliferation and colonyformation of leukemia cell line K562

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Abstract:

Objective: To explore the role of membranebound stem cell factor (mSCF) in the pathogenesis of leukemia. Methods: The eukaryotic expression plasmid of soluble stem cell factor（sSCF）precursor (pTARGETs) was constructed. Overlap PCR was used to obtain mSCF sequence with the deletion of exon 6, and pTARGETm was constructed. After verified by DNA sequencing, pTARGETm and control pTARGET vector were transfected into K562 cells via Lipofetamine 2000, and the positive cells were screened by G418. K562 cells stably transfected with pTARGETm were verified by RTPCR and Western blotting. Proliferation and colonyformation of these stably transfected cells were studied. Results: The sSCF and mSCF eukaryotic expression vectors were successfully constructed. The stable transfectants K562V, K562S, and K562M were obtained. Under Ubottom culture condition, proliferation ability of K562M cells was significantly stronger than those of K562V or K562S (both P<0.01). Colonyformation ability of K562M was significantly higher than those of K562V and K562S (both P<0.01). Furthermore, the colony size of K562M was larger than those of the other two kinds of cells. Conclusion: mSCF significantly enhances proliferation and colonyformation of leukemia cells by a juxtacrine mechanism.

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Supported by the Major State Basic Research Development Program of China (2008CB517301); China Medical Board (CMB) (No.A2007001)

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