| 丁媛 吴秀娟 向芳 康晓静 于世荣 冯燕艳 王红娟 普雄明.miR-181b-5p 对于卡波西肉瘤细胞SLK细胞增殖和凋亡的影响[J].现代生物医学进展英文版,2015,15(21):4005-4008. |
| miR-181b-5p 对于卡波西肉瘤细胞SLK细胞增殖和凋亡的影响 |
| The Effects of MicroRNA-181b-5p on Proliferation and Apoptosis of HumanKaposi's Sarcoma Cell Line SLK |
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| DOI： |
| 中文关键词: miR-181b-5p 卡波氏肉瘤 增殖 凋亡 |
| 英文关键词: miR-181b-5p Kaposi's sarcoma Proliferation Apoptosis |
| 基金项目:国家自然科学基金地区科学基金项目（81260311）；新疆维吾尔自治区自然科学基金项目（2014211A059） |
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| 中文摘要: |
| 目的：探讨Dppa2 基因5' 端启动子区Oct4 结合位点突变对Dppa2基因启动子活性的影响。方法：PCR 扩增包括Oct4 结合
位点的Dppa2 基因5' 端转录起始点上游-2439～+293 bp 的启动子序列，片段长度为2732 bp。将该片段连接到pGL3-Basic 载体，
构建野生型pGL3-2439表达载体。采用定点突变法，将-1959～-1957 位碱基的GCA突变成TAG，构建Oct4 结合位点突变型
pGL3-mo2439 表达载体。用上述两种表达载体、PGL3-basic 载体和Oct4 表达载体分别瞬时转染HEK 293 细胞。细胞培养48 h
后，利用双荧光素酶报告系统测定各组细胞表达的荧光素酶的相对活性。结果：经琼脂糖凝胶电泳及测序鉴定，证实野生型
(pGL3-2439)和突变型(pGL3-mo2439)载体构建成功。荧光素酶活性测定结果显示，转染Dapp2 基因启动子野生型pGL3-2439 表
达载体的细胞组荧光素酶的相对活性为16.307，突变型pGL3-mo2439 表达载体的细胞组荧光素酶的相对活性为10.634。Oct4 结
合位点突变后，Dppa2 基因启动子区转录活性较野生型降低了35 %。结论：Dppa2基因5'端启动子区-1959～-1957 位的Oct4 结
合位点突变可能导致Dppa2 基因启动子活性下降。 |
| 英文摘要: |
| Objective:miR-181b-5p mimics and inhibitors were transfected into Kaposi's sarcoma cell line SLK through liposomal
transfection. The effect of miR-181b-5p on SLK cell proliferation and apoptosis of biological activities were studied to provide theoretical
basis for further investigation on the role of miR-181b-5p in the pathogenesis of Kaposi's sarcoma.Methods:Human Kaposi's sarcoma
cell line SLK was transfected by liposomes. The proliferation of SLK cell was assessed by MTT assay; apoptosis rates of each group
were determined by flow cytometry at 48 hours after transfection.Results:Compared to the negative control group, proliferative capacity
of miR-181b-5p mimics has increased significantly after transfection (P＜0.01), proliferative capacity of miR-181b-5p inhibitor has decreased
significantly after transfection (P＜0.01). Flow cytometry showed that, apoptosis rate of the SLK cells has decreased significantly
after transfection with miR-181b-5p mimics (5.5± 0.6)%compared to that of corresponding negative control (7.6± 0.4)% (P＜0.01).
Apoptosis rate of the SLK cells has increased significantly after transfection with miR-181b-5p inhibitor (14.8± 1.0)compared to that of
corresponding negative control(7.8± 0.5) % (P＜0.01).Conclusion:Transfection with miR-181b-5p mimics/inhibitors significantly affected
SLK cell proliferation and apoptosis. Further researches on biological characteristics of miR-181b-5p and validation of the associated
target genes are therefore of great significance. |
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