中华皮肤科杂志 ›› 2007, Vol. 40 ›› Issue (3): 170-172.

• 论著 • 上一篇    下一篇

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

于建斌1, 张岚2, 高冬玲2, 李琳1, 陈燕辉2, 陈奎生1, 张云汉2   

  1. 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、河南省肿瘤病理重点实验室, 450052
  • 收稿日期:2006-03-28 出版日期:2007-03-15 发布日期:2007-03-15
  • 通讯作者: 张云汉,email:yhzhang@zzu.edu.cn E-mail:yhzhang@zzu.edu.cn

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

YU Jian-bin1, ZHANG Lan2, GAO Dong-ling2, LI Lin1, CHEN Yan-hui2, CHEN Kui-sheng1, ZHANG Yun-han2   

  1. Department of Dermatology, First Affiliated Hospital of Zhengzhou University Zhengzhou 450052, China
  • Received:2006-03-28 Online:2007-03-15 Published:2007-03-15

PDF (PC)

40

赞

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24 h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48 h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.

关键词: Marshall-White综合征, 黑色素瘤, 肝素裂合酶, 寡核昔酸类,反义

Abstract: Objective To study the effects of heparinase antisense oligodeoxynucleotide(ASODI)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.

Key words: Melanoma, Heparin lyase, Oligonuclcotidcs, antisense