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Inhibitory effect of human melanoma differentiation associated gene7 on chronic myelocytic leukemia cell line K562
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Abstract:
Abstract Objective: To explore the inhibitory effect of human melanoma differentiation associated gene7 (mda7/IL24) on chronic myelocytic leukemia cell line K562 in vitro and in vivo.Methods: The expression of mda7〖STBZ〗 receptor was examined in 11 malignant hematopoietic cell lines（NB4, HL60, CEM, Namalwa, Jurkat, KG1a, U937, K562, Ramos, J61, and HEL） by RTPCR. Then the constructed pTargetIL24 eukaryotic expression vector was transduced into K562 cells via Lipofectamine reagent. The expression of mda7 mRNA and protein was verified by RTPCR and Western blotting. MTT assay, colony forming assay, flow cytometry, AnnexinⅤ/PI and xenograft tumor models in nude mice were used to assess the effects of mda7 on tumor cells proliferation, colony forming, cell cycle, apoptosis, and tumorigenesis, respectively. Results: The expression of mda7 intact receptor was not detected in the 11 malignant hematopoietic cell lines. Significant expression of mda7 mRNA and protein was found in K562 cells stably transfected with mda7. Compared with control cells transfected with plasmid vector or untransfected cells, cells transfected with mda7had decreased tumor cells proliferation (P<0.05), inhibited colony formation (P<0.05), and more cells were arrested in G0/G1 stage (P<0.05). However, there was no significant difference in cells apoptosis between control cells and K562 cells transfected with mda7. Tumor xenograft models in BALB/c nude mice showed that mda7/IL24 significantly inhibited the growth of K562 transplantation tumor (P<0.01). Conclusion: Human mda7/IL24 can efficiently inhibit the proliferation of chronic myelocytic leukemia cell line K562 in vitro and in vivo, possibly by inducing K562 cell arrest in G0/G1 stage.
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Project Supported:
Supported by the National Natural Science Foundation of China (No. 30672364, No.30872983); the Key Program of Applied Basic Research Foundation of Tianjin (No. 07JCZDJC07600)
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