bcl-2硫代反义寡核苷酸抑制人恶性黑色素瘤A375细胞bcl-2mRNA及蛋白的表达

Expression of bcl-2 mRNA and protein suppressed by antisense oligodexyonucleotide with phosphorothiote-modification in human melanoma A375 cells line

目的 观察bcl-2硫代反义寡核苷酸（ASODN）对人恶性黑色素瘤A375细胞bcl-2mRNA及蛋白表达的影响。方法 对ASODN进行硫代修饰、脂质体转染人黑色素瘤A375细胞。实验分反义ASODN组、正义SODN组和对照组，采用逆转录聚合酶链反应（RT—PCR）和免疫组织化学检测bcl-2mRNA及蛋白水平。结果 免疫组织化学检测显示，反义组Bcl-2蛋白表达水平显著低于对照组和正义组（分别为53．14±4．26、138．22±8．45和141．08±7．83，P〈0．01）；RT—PCR结果示，反义组bcl-2mRNA水平明显低于对照组和正义组（分别为0．38±0．1l、0．964-0．13和O．97±0．14，P〈0．01）。结论 bcl-2硫代反义寡核苷酸能下调人黑色素瘤A375细胞bcl-2mRNA基因水平，阻断蛋白表达。

Objective To observe the effect of bcl-2 antisense oligodexyonucleotide （ASODN） by phosphorothiote-modification on the expression of bcl-2 mRNA and protein in human melanoma A375 cells line. Methods With the phosphorothiote-modification and liposome-encapsulation of ASODN, A375 cells were divided into ASODN group, nonsense oligodexyonucleotide （SODN） group and control group. Bcl-2 mRNA and protein were detected by RT-PCR and immunohistochemistry, respectively. Results The expression of bcl-2 protein was remarkably decreased in ASODN group than that in SODN group and control group （53.14 ±4.26 vs 138.22 ± 8.45, 53.14 ±4.26 vs 141.08± 7.83, both P 〈 0.01 ）. The level of bcl-2 mRNA was significantly lower than that in SODN group and control group （ 0.38 ± 0. 11 vs 0.96 ± 0.13, 0.38±0.11 vs 0.97 ±0. 14, both P 〈0.01）. Conclusion Bcl-2 antisense oligodexyonucleotide could down-regulate the bcl-2 level and block its protein expression.