特异性反义寡核苷酸抑制白血病细胞株HLA－DPB_1基因表达

INHIBITION OF HLA-DPB1GENE EXPRESSION BYANTISENSE OLIGONUCLEOTIDES IN LEUKEMIA CELLLINES

反义寡核苷酸可以特异性地抑制基因表达，将ＨＬＡ－ＤＰＢ＿１特异性反义寡核苷酸加入细胞培养基中，对ＨＬ－６０和Ｋ５６２白血病细胞株由γ－干扰素诱导的ＨＬＡ－ＤＰＢ＿１表达有特异性抑制作用。当反义寡核苷酸浓度在４０μｇ／ｍｌ时，１小时后即出现ＨＬＡ－ＤＰＢ＿１基因表达抑制现象，持续时间达４８小时，７２小时后表达又逐渐恢复；反义寡核苷酸抑制基因表达有很高的特异性，在ＨＬＡ－ＤＰＢ＿１表达完全被封闭的情况下，与它链锁紧密的ＤＱＢ＿１、ＤＲＢ＿１表达依然如常。另外，不同浓度的反义寡核苷酸，从５μｇ／ｍｌ到６０μｇ／ｍｌ，对两种细胞株￣３Ｈ－ＴｄＲ掺入无明显影响，说明它对细胞生长影响不大。

ntisense oligonucleotides of HLA-DPB1 second ex-tron could inhibit HLA-DPB1 expression in HL-60 andK562 cell lines. When the cell lines were exposed to theantisenee oligonucleotides (40μg/ml in culture medial),the inhibition of DPB1 gene expression could continuefor 48 hours, and in 72 hours the gene expression re-turned to the untreated level. While the expression ofHLA-DPB1 gene was suppressed undetectable, the ex-prestsion of HLA-DRB1 and DQB1 showed no significantchanges, suggesting that the antisense oligonucleotidesare highly specific. In addition, different concentrationsof antisense oligonucleotides did not affect the incorpora-tion of ￣3H-TdR, indicating that they did not interfere theproliferation of the cell lines.