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目的合成针对长型ＰＭＬＲＡＲα融合基因ｍＲＮＡ起始部位和融合位点的反义核酸（ＡＳ），探讨ＡＳ的稳定性、特异性及其对ＮＢ４细胞生长、凋亡的影响。方法以ＮＢ４细胞株为研究对象，采用台盼蓝拒染法进行细胞计数；聚丙烯酰胺凝胶进行寡核苷酸的电泳；流式细胞仪、ＤＮＡ电泳、细胞荧光染色进行细胞凋亡检测。结果合成的ＡＳ稳定、耐细胞内外核酸酶、无非特异性作用；起始部位ＡＳ（ＳＴＡＳ）和融合位点ＡＳ（ＦＵＡＳ）明显抑制细胞增殖，且呈剂量依赖性。浓度２０μｇ／ｍｌ时增殖抑制率０％～１１．３％；８０μｇ／ｍｌ时，抑制率５０．０％～６７．７％。ＦＵＡＳ（６０μｇ／ｍｌ）处理第７天、第９天出现明显的凋亡细胞群，处理后３天、５天、７天、９天的凋亡细胞率分别为９．３％、２４．５％、４１．０％、３４．２％。结论针对长型ＰＭＬＲＡＲα融合基因的ＡＳ设计、合成是成功的；抗ＰＭＬＲＡＲαＡＳ能抑制细胞增殖，诱导细胞凋亡。
Objective To synthesize antisense oligonucleotide (AS) targeting the initiation site and fusion site of long PMLRARα fusion gene, and to investigate the stability and specificity of AS and its effect on the growth and apoptosis of NB4 cells. Methods The NB4 cell line was used as the research object. Cell counting was performed by trypan blue exclusion method. Polyacrylamide gel electrophoresis was performed by using oligonucleotide. Cell apoptosis was detected by flow cytometry, DNA electrophoresis and fluorescence staining. Results The AS was stable and resistant to non-specific endonucleases. ASAS (STAS) and AS (FUAS) inhibited the cell proliferation in a dose-dependent manner. When the concentration was 20μg / ml, the proliferation inhibition rate was 0% ~ 11.3%. When the concentration was 80μg / ml, the inhibition rate was 50.0% ~ 67.7%. FUAS (60μg / ml) for 7 days, the apoptotic cell population appeared on the 9th day. The apoptotic cell rates at 3, 5, 7 and 9 days after treatment were 9.3% and 24.5 %, 41.0%, 34.2%. Conclusion AS design and synthesis of long PMLRARα fusion gene were successful. AntiPMLRARαAS could inhibit cell proliferation and induce apoptosis.