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目的人白血病细胞系ＨＬ－６０细胞高表达凋亡抑制基因ｂｃｌ－２蛋白，是研究细胞增殖、凋亡的良好模型。通过脂质体转染ｂｃｌ－２反义核酸，观察其对细胞增殖的抑制，对ｂｃｌ－２蛋白表达影响，及诱导细胞凋亡出现。方法ＨＬ－６０细胞在３７℃、５％ＣＯ２条件下，培养于含１５％小牛血清及含青、链霉素（每毫升各１００Ｕ）的ＲＰＭＩ１６４０完全培养基中，细胞接种密度为１０７／Ｌ，反义核酸终浓度为５μｍｏｌ／Ｌ，反义核酸与脂质体比例为１．０ＯＤ：５０μｌ。脱离小牛血清转染２４ｈ，同时设置空白组、无义组、反义组、脂质体组、脂质体＋无义组，再连续培养４天，做免疫组化，检测ｂｃｌ－２蛋白表达。提取总ＲＮＡ，行ＲＴ－ＰＣＲ分析ｂｃｌ－２ｍＲＮＡ表达，以及透射电镜检查，观察细胞凋亡情况及脂质体转染情形。结果脂质体转染ｂｃｌ－２反义核酸的作用：（１）抑制ＨＬ－６０细胞增殖，在转染后第五天最明显，经统计学处理，反义组与空白及无义对照组有显著性差别，脂质体转染反义核酸组与其余组均有统计上显著性差别。（２）降低ｂｃｌ－２蛋白表达水平，处理前ｂｃｌ－２免疫组化阳性率为６２±１．４８％，经转染ｂｃｌ－２反义核酸处理后，ｂｃｌ－２表达率为３２．５±１．?
The target human leukemia cell line HL-60 cells highly express apoptosis-inhibiting gene bcl-2 protein, which is a good model to study cell proliferation and apoptosis. Bcl-2 antisense oligonucleotides were transfected by lipofectamine to observe the effect of bcl-2 antisense RNA on bcl-2 protein expression and cell apoptosis. Methods HL-60 cells were cultured in complete medium of RPMI1640 containing 15% fetal bovine serum and streptomycin (100U / ml each) under the conditions of 37 ℃ and 5% CO2. The cell seeding density was 107 / L , Antisense nucleic acid final concentration of 5μmol / L, antisense nucleic acid and liposome ratio 1.0OD: 50μl. After transfection for 24 hours, the blank control group, non-sense group, antisense group, lipofectamine group and liposome + non-sense group were transplanted for 24 hours. After 4 days of continuous culture, immunohistochemistry and bcl-2 protein expression. The total RNA was extracted and the expression of bcl-2 mRNA was analyzed by RT-PCR. The apoptosis and the transfection of liposome were observed by transmission electron microscopy. Results Liposome transfected bcl-2 antisense nucleic acid: (1) Inhibition of HL-60 cell proliferation, the most obvious on the fifth day after transfection, the statistical analysis, antisense group and blank control group There was a significant difference between the liposome transfected antisense nucleic acid group and the remaining groups were statistically significant differences. (2) The expression of bcl-2 protein was decreased, the positive rate of bcl-2 immunohistochemistry before treatment was 62 ± 1.48%, and the bcl-2 expression rate was 32.5 ± 1. ?