Enhancing effect of DLL3 gene silencing on the sensibility in human leukemia K562/ADM cells to adriamycin

WANG Shi-guang* SI Xu-yan WANG Li-jun TONG Lei LI Yang-jie

doi:10.16098/j.issn.0529-1356.2018.05.010

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Acta Anatomica Sinica ›› 2018, Vol. 49 ›› Issue (5) : 624-629. DOI: 10.16098/j.issn.0529-1356.2018.05.010

Cancer Biology

Enhancing effect of DLL3 gene silencing on the sensibility in human leukemia K562/ADM cells to adriamycin

WANG Shi-guang ^1* SI Xu-yan² WANG Li-jun¹ TONG Lei³ LI Yang-jie⁴

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Abstract

Objective To investigate whether Delta-like ligand 3 (DLL) gene silencing could influence the sensibility of human leukemia K562/ADM cells to adriamycin (ADM) in vitro . Methods K562/ADM cells were stably transfected with specific shRNA interference plasmid vector targeting for DLL3. The mRNA expression level of DLL3 were measured by RT-PCR. CCK-8 assay was employed to detect the cytotoxic effect of ADM in K562 and K562/ADM cells. The apoptosis and intracellular ADM concentration were analyzed by flow cytometry. Western blotting was performed to determine the protein expression levels of DLL3, glutathione S transferases-π, (GST-π) and P-glycoprotein (P-gp). Results K562/ADM cells had a significantly higher mRNA and protein expression level of DLL3 than K562 cells (P<0.05). The IC₅₀ value of K562 and K562/ADM cells to ADM were 1.08 and 34.93 mg/L respectively. After DLL3 gene silencing, the resistant factor of K562/ADM cells declined to 13.12 with a reversal fold of 2.47. DLL3 gene silencing had no effect on apoptosis of K562/ADM cells, but a significant effect on downregulating protein levels of P-gp and GST-π (P<0.05) and increasing intracellular ADM concentration (P<0.05), and resulting in an enhance of ADM-induced apoptosis (P<0.05). Conclusion DLL3 gene silencing may enhance the sensibility of K562/ADM cells to ADM by down-regulating P-gp and GST-π, and increasing intracellular ADM concentration.

Key words

DLL3 gene / K562/ADM cell / P-glycoprotein / Glutathione S transferases-π / RT-PCR / Leukemia / Human

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WANG Shi-guang SI Xu-yan WANG Li-jun TONG Lei LI Yang-jie. Enhancing effect of DLL3 gene silencing on the sensibility in human leukemia K562/ADM cells to adriamycin[J]. Acta Anatomica Sinica. 2018, 49(5): 624-629 https://doi.org/10.16098/j.issn.0529-1356.2018.05.010

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