Construction of artificial microRNA vector interfering mouse Cbfβ gene and establishment and identification of stable cell line

黄科; 胡宗凯; 喻胜鹏; 傅景曙; 李伟; 董世武; 谢肇

您当前的位置：

首页 >

文章列表页 >

Construction of artificial microRNA vector interfering mouse Cbfβ gene and establishment and identification of stable cell line

更新时间：2026-01-09

- Construction of artificial microRNA vector interfering mouse Cbfβ gene and establishment and identification of stable cell line
- Journal of Army Medical University Vol. 36, Issue 7, Pages: 682-686(2014)
- 作者机构：
- 作者简介：
- 基金信息：
- DOI：
  CLC： R394-33;R394.2
- Published：2014
- 稿件说明：
移动端阅览
黄科,胡宗凯,喻胜鹏,傅景曙,李伟,董世武,谢肇.Cbfβ人工miRNA干扰载体构建及稳定细胞株系的获得与鉴定[J].陆军军医大学学报(原第三军医大学学报),2014,36(07):682-686.

　Huang Ke,Hu Zongkai,Yu Shengpeng,Fu Jingshu,Li Wei,Dong Shiwu,Xie Zhao.Construction of artificial microRNA vector interfering mouse Cbfβ gene and establishment and identification of stable cell line[J].J Amry Med Univ (J Third Mil Med Univ),2014,36(07):682-686.
黄科,胡宗凯,喻胜鹏,傅景曙,李伟,董世武,谢肇.Cbfβ人工miRNA干扰载体构建及稳定细胞株系的获得与鉴定[J].陆军军医大学学报(原第三军医大学学报),2014,36(07):682-686. DOI：

　Huang Ke,Hu Zongkai,Yu Shengpeng,Fu Jingshu,Li Wei,Dong Shiwu,Xie Zhao.Construction of artificial microRNA vector interfering mouse Cbfβ gene and establishment and identification of stable cell line[J].J Amry Med Univ (J Third Mil Med Univ),2014,36(07):682-686. DOI：

摘要

目的构建Cbfβ基因干扰载体，获得Cbfβ人工miRNA干扰载体和稳定细胞株系，检测其对间充质干细胞C3H10T1/2细胞成骨分化的影响。方法设计特异性干扰小鼠Cbfβ基因的人工miRNA序列，构建Cbfβ干扰载体Blockit miRNA-cbfβ-miRNA1/miRNA2，将其转入C3H10T1/2细胞获得稳定细胞株；用BMP-2诱导3 d后采用荧光定量PCR(qRT-PCR)和Western印迹检测Cbfβ、Runx2和成骨标志基因ALP表达水平。结果成功构建成骨转录因子Cbfβ的干扰载体并获得了稳定细胞株系，成骨标志基因ALP和Cbfβ的mRNA和蛋白表达水平明显下降（

0.05），而Runx2下降不显著（

0.05）。结论 miRNA-cbfβ-miRNA1/miRNA2载体可以抑制成骨转录因子Cbfβ的功能，进而降低成骨标志基因ALP蛋白和mRNA的表达，并获得可以影响成骨过程的稳定细胞株系。

Abstract

Objective To construct and identify miRNA expression vectors interfering mouse core binding factor β subunit (Cbfβ) gene and to determine their effects on osteogenic differentiation of mesenchymal stem cell C3H10T1/2. Methods Artificial microRNA sequences specifically interfering mouse Cbfβ gene were designed and Blockit miRNA-Cbf-β-miRNA1/miRNA2 vectors interfering mouse Cbfβ gene were constructed. The vectors were transferred into C3H10T1/2 cells to get stable cell line. Quantitative RT-PCR and Western blotting were used to detect the expression levels of Cbfβ

Runt-related tra

nscription factor 2 (Runx2) and osteogenesis marker gene alkaline phosphatase (ALP) in the cells in 3 d after BMP-2 treatment. Results Artificial microRNA vectors interfering Cbfβ were successfully constructed and stable cell line was obtained. The expression level of ALP and Cbfβ were obviously reduced （

0.05）

while Runx2 expression was not notably decreased （

0.05）. Conclusion miRNA expression vectors miRNA-Cbf-β-miRNA1/miRNA2 targeting Cbfβ gene may inhibit function of osteogenic transcription factor Cbfβ and then decrease protein and mRNA expression of ALP. Stable cell line with influenced osteogenic ability is obtained.

关键词

Keywords

references

Views

下载量

CSCD

Alert me when the article has been cited

提交

Tools

Publicity Resources

Protective effect of placenta-derived mesenchymal stem cell transplantation against severe acute pancreatitis in rats

Expression difference and mineralization correlation of p75NTR in rat ectomesenchymal stem cells at different fusion states

Effect of periostin on proliferation of keloid mesenchymal stem cells in vitro

Expression of ferritin heavy chain 1 in neural differentiation of bone marrow mesenchymal stem cells

Poly(I:C)-pretreated MSCs protect intestinal barrier function in rats after sepsis

Related Author

黄启林

杨屹

罗晨李帅

汤礼军

孙红玉

肖勇

李刚

王莹莹

Related Institution

No data

⁰