Abstract
Objective: To investigate the effects of anti-PML (promyelocytic leukemia) or anti-PML/RARα (promyelocytic leukemia/retionic acid receptorα) antisense oligonucleotides on cell growth, expression of PML-RARα mRNA and PML-RARα/PML protein location of NB4 cell lines. Methods: RT-PCR was used for detecting PML-RARα mRNA expression, trypan blue exclusion for cell count, methylcellose assay for leukemic colony forming unit detection, immunofluorescence for PML-RARα/PML protein location. Results: Both anti-PML start codon region antisence (STAS) and anti-PML-RARα fusion region antisence (FUAS) could inhibit cell growth and the formation of acute myelocytic colony forming unit of cells(AML-CFU). Cells become partial differentiated at days 5, being more obvious in FUAS-treated cells than in STAS ones. Down regulation of PML-RARα mRNA expression occurred at 24 hours in STAS and FUAS-treated cells and maintained for up to 72 hours. Immuno-fluorescence analysis with anti-PML monoclonal antibody showed a remarkable decrease even complete disappearance of microgranules. The residual granules became enlarged as discrete dots (<10 per cell), similar to normal POD structure in some STAS-treated cells at 24 hours. At 72 hours, nearly all the granules disappeared. Similar changes were observed in FUAS-treated cells. Conclusion: Both PML and PML-RARα antisence oligonucleotides can specially block the expression of PML-RARα at mRNA and protein levels. PML protein is implicated in the regulations of cell differentiation.
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Foundation item: This work was supported by the National Natural Science Foundation of China(No. 39590291).
Now works in Affiliated hospital of Nan Tong Medical College, Nantong 226001, China;
Biography: Chen Ye, (1962–), doctor of medicine, vice-chief physician and associate professor, majors in pathogenesis and treatment of leukemia.
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Chen, Y., Miao, Jm., Fang, Zw. et al. Effects of PML and PML/RMRα antisence oligonucleotide on promyelocytic leukemia cell NB4. Chin. J. Cancer Res. 13, 92–95 (2001). https://doi.org/10.1007/s11670-001-0022-z
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DOI: https://doi.org/10.1007/s11670-001-0022-z