2012Chinese Journal of NeuromedicineRequires access

Small interfering RNA -mediated cathepsin D knock-down impairing biological behavior of glioma cells

Jinqiao Zhou, Jingtao Wang, Qiu-hong Liu, Xin-bing Guo, Laijun Song

Open publisher page 0 citations

Abstract

Objective To explore the effect of small interfering RNA (siRNA)-mediated cathepsin D (CD) knock-down on biological behavior of U87 cells. Methods CD-siRNA,scramble siRNA and control siRNA mediated by lipofectamin were introduced into the U87 cells,respectively.The silencing effect of CD was verified by RT-PCR and Western blotting in terms of CD mRNA and protein expressions. The cell viability of scramble and CD siRNA treated U87 cells was measured by MTT on 1-5 d after transfection. The invasive potential of scramble and CD siRNA treated U87 cells was measured by Transwell coated with Matrigel 48 h after transfection. Results Forty-eight h after transfection, mRNA and protein expressions of CD in CD siRNA treated U87 cells were remarkably reduced as compared with those in the scramble siRNA and control siRNA treated U87 cells (P<0.05).The absorbance (A value) of CD siRNA treated U87 cells was significantly lower than that in the scramble siRNA treated U87 cells 2,3,4 and 5 d after transfection (P<0.05); and the invasive potential of CD siRNA treated U87 cells was suppressed significantly as compared with that in the scramble siRNA treated U87 cells (P<0.05). Conclusion CD has the potential to be a promising therapeutic target. Key words: Glioma; Small interfering RNA; Cathepsin D

About this research paper

What this paper is about

Objective To explore the effect of small interfering RNA (siRNA)-mediated cathepsin D (CD) knock-down on biological behavior of U87 cells. Methods CD-siRNA,scramble siRNA and control siRNA mediated by lipofectamin were introduced into the U87 cells,respectively.The silencing effect of CD was verified by RT-PCR and Western blotting in terms of CD mRNA and protein expressions. The cell viability of scramble and CD siRNA treated U87 cells was measured by MTT on 1-5 d after transfection. The invasive potential of scramble and CD siRNA treated U87 cells was measured by Transwell coated with Matrigel 48 h after transfection. Results Forty-eight h after transfection, mRNA and protein expressions of CD in CD siRNA treated U87 cells were remarkably reduced as compared with those in the scramble siRNA and control siRNA treated U87 cells (P<0.05).The absorbance (A value) of CD siRNA treated U87 cells was significantly lower than that in the scramble siRNA treated U87 cells 2,3,4 and 5 d after transfection (P<0.05); and the invasive potential of CD siRNA treated U87 cells was suppressed significantly as compared with that in the scramble siRNA treated U87 cells (P<0.05). Conclusion CD has the potential to be a promising therapeutic target. Key words: Glioma; Small interfering RNA; Cathepsin D

Why it matters

A significance statement is not available in the OpenAlex record.

Key contribution

A contribution statement is not available in the OpenAlex record.

Method / approach

Method details are not available in the OpenAlex metadata.

Main findings

Findings are not separately available in the OpenAlex metadata.

Limitations

Limitations are not available in the OpenAlex metadata.

Applications

Application details are not available in the OpenAlex metadata.

Available abstract

Objective To explore the effect of small interfering RNA (siRNA)-mediated cathepsin D (CD) knock-down on biological behavior of U87 cells. Methods CD-siRNA,scramble siRNA and control siRNA mediated by lipofectamin were introduced into the U87 cells,respectively.The silencing effect of CD was verified by RT-PCR and Western blotting in terms of CD mRNA and protein expressions. The cell viability of scramble and CD siRNA treated U87 cells was measured by MTT on 1-5 d after transfection. The invasive potential of scramble and CD siRNA treated U87 cells was measured by Transwell coated with Matrigel 48 h after transfection. Results Forty-eight h after transfection, mRNA and protein expressions of CD in CD siRNA treated U87 cells were remarkably reduced as compared with those in the scramble siRNA and control siRNA treated U87 cells (P<0.05).The absorbance (A value) of CD siRNA treated U87 cells was significantly lower than that in the scramble siRNA treated U87 cells 2,3,4 and 5 d after transfection (P<0.05); and the invasive potential of CD siRNA treated U87 cells was suppressed significantly as compared with that in the scramble siRNA treated U87 cells (P<0.05). Conclusion CD has the potential to be a promising therapeutic target. Key words: Glioma; Small interfering RNA; Cathepsin D

Key concepts: Transfection, Small interfering RNA, Chemistry, Gene silencing, Molecular biology, Matrigel, Gene knockdown, Glioma

Related papers

Back to paper searchBrowse research topicsOriginal source
Small interfering RNA -mediated cathepsin D knock-down impairing biological behavior of glioma cells — Research Paper | ScholarLens