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[Expression of interleukin-18 in human leukemia cell line J6-1 and its significance]
1Institute of Hematology, Chinese Academy of Medical Sciences and Peking Union Medical College. Tianjin 300020, China.
Objective:
To elucidate the expression and regulation of interleukin-18 (IL-18) in human leukemia cell line J6-1, and investigate its possible role in leukemogenesis.
Methods:
IL-18 mRNA expression was detected using reverse transcriptase-ploymerase chain reaction (RT-PCR), and the specificity of IL-18 cDNA was verified by restriction endonucleases analysis. To sequence and analyze its cDNA alignment, a recombinant plasmid of IL-18 was constructed. Employing the antisense technique, we ionvestigated the effects of IL-18 antisense oligodeoxynucleiotide (ASODN) on the proliferation of J6-1 cells.
Results:
J6-1 cells constitutively expresses IL-18 mRNA, and its cDNA sequence shared 99% homology with the published IL-18 sequence,except that a synonymous point mutation (Codon35 UCA to UCC) was confirmed by sequencing and alignment analysis. Peripheral blood mononuclear cells (PBMC) of healthy volunteers weakly expresses IL-18 mRNA (0.13 +/- 0.05), while CpG-ODN significantly up-modulated IL-18 expression in PBMC (0.82 +/- 0.24), but not in J6-1 cells. Compared with K562, HL-60, U937 and LCL cells, IL-18 mRNA level expression of in J6-1 cells was the highest (0.98 +/- 0.29). ASODN of IL-18 markedly inhibited the proliferation of J6-1 cells (inhibition rate 43.3%), but did not show significant effects on HL-60 cells (inhibition rate 16.7%).
Conclusions:
J6-1 cells constitutively expresses IL-18 mRNA, and no significant mutation was detected coding region of IL-18 cDNA. IL-18 might be a positive regulation factor for the proliferation of J6-1 cells after in an autocrine model.