反义寡核苷酸抑制人恶性黑素瘤A375细胞乙酰肝素酶mRNA及其蛋白的表达  

Inhibitory effects of antisense oligodeoxynucleotide on the expression of heparinase mRNA and protein in human malignant melanoma cell line A375

作　　者：于建斌[1] 张岚[2,3] 高冬玲[2,3] 李琳[1] 陈燕辉[1] 陈奎生[2,3] 张云汉[2,3] 

机构地区：[1]郑州大学第一附属医院皮肤科 ,450052 [2]郑州大学第一附属医院病理科 ,450052 [3]河南省肿瘤病理重点实验室

出　　处：《中华皮肤科杂志》2007年第3期170-172,共3页Chinese Journal of Dermatology

摘　　要：目的 研究乙酰肝素酶反义寡核苷酸（ASODN）对人恶性黑素瘤A375细胞乙酰肝素酶mRNA及其蛋白表达的影响。方法 分空白组、无关序列（N-ODN）组和反义（ASODN）组。以脂质体包埋后分别转染人恶性黑素瘤细胞株A375，采用原位杂交、免疫组化法检测乙酰肝素酶mRNA及其蛋白表达的变化。结果 转染24h后，ASODN组A375细胞乙酰肝素酶mRNA表达较对照组、N-ODN组均显著下调（P均〈0．01）；转染48h后，与对照组、N-ODN组相比，ASODN组A375细胞的乙酰肝素酶蛋白水平均显著下降（P均〈0．01）。结论 乙酰肝素酶ASODN可下调A375细胞乙酰肝素酶mRNA及其蛋白的表达。Objective To study the effects of heparinase antisense oligodeoxynucleotide （ ASODN ） on the expression of heparinase mRNA and protein in human malignant melanoma cell line A375. Methods Heparinase ASODN and its controlled nonsense oligodeoxynucleotide （N-ODN） were designed and synthesized. A375 cells were divided into three groups： ASODN group, transfected with 30 μmol/L of heparinase ASODN and lipofectin; N-ODN group, transfected with 30 μmol/L of heparinase N-ODN and lipofectin; blank control group. Heparinase mRNA and protein were detected by in situ hybridization and immunohistochemistry, respectively. Results At 24 h after the transfection, the expression of heparinase mRNA was significantly lower in the ASODN group than that in the N-ODN group and control group （ 1.250 ±0.500 vs 4.000 ± 0.817, 1.250 ±0.500 vs 4.250 ±0.500, both P〈 0.01 ）. The expression of heparinase protein was also significantly decreased in the ASODN group than that in the control and N-ODN group at 48 h after transfection （ 5.000 ±0.817 vs 1.500± 0.577, 4.500 ± 0.577 vs 1.500 ±0.577, both P 〈 0.01 ）. Conclusion Heparinase ASODN could down-regulate the expression of heparinase mRNA and protein in A375 cells.

关 键 词：黑色素瘤 肝素裂合酶 寡核苷酸类 反义 

分 类 号：R686[医药卫生—骨科学]