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Updated: Feb 17, 2026
Isolation and Flow Cytometric Analysis of Glioma-infiltrating Peripheral Blood Mononuclear Cells
Published on: November 28, 2015
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1The 1st Department of Neurosurgery, The People's Hospital of Yinan, Yinan, Shandong, China. Zhenyanlee@hotmail.com.
Objective:
Gliomas are accompanied with high mortality owning to their invasive peculiarity and vulnerability to drug resistance. miR-21 is a vital oncogenic miRNA that regulates drug resistance of tumor cells. This study aims to elucidate the function of miR-21 in human glioma cells resistant to carmustine (BCNU) and to demonstrate the underlying molecular mechanism.
Materials And Methods:
BCNU-sensitive cells (SWOZ2 cells) were transfected with miR-21 agomir and negative control, and BCNU-resistance cells (SWOZ2-BCNU cells) were transfected with miR-21 antagomir and negative control. The Real-time fluorescence quantitative PCR was used to detect and compare the levels of miR-21expression between SWOZ2-BCNU and SWOZ2 cells. The drug sensitivity of these cells to BCNU was determined by Cell Counting Kit-8 (CCK-8) assay. The protein expression of Spry2 was detected by Western blotting.
Results:
The expression level of miR-21 was remarkably higher in SWOZ2-BCNU cells than that in SWOZ2 cells. The half-maximal inhibitory concentration (IC50) of BCNU was obviously higher for SWOZ2-BCNU cells than that for SWOZ2 cells. Besides, we found that aberrant expression of miR-21 in SWOZ2-BCNU cells is responsible for glioma BCNU-resistance. Consistently, Spry2 protein levels were significantly reduced in SWOZ2-BCNU as well as in miR-21 agomir-transfected cells, inversely correlated to miR-21 expression. The results of si-Spry2 co-transfection suggested that the effect of miR-21 on glioma BCNU-resistance is mediated through Spry2.
Conclusions:
miR-21 enhances the resistance of human glioma cells to BCNU by decreasing the expression of Spry2 protein. Thus, Spry2 may be a novel therapeutic target for treating glioma BCNU-resistance.
Insights
MicroRNA-21 (miR-21) increases resistance in human glioma cells to carmustine (BCNU) by reducing Spry2 protein levels. Targeting Spry2 may offer a new therapeutic strategy for BCNU-resistant gliomas.
Area of Science:
- Oncology
- Molecular Biology
- Genetics
Background:
- Gliomas exhibit high mortality due to invasiveness and drug resistance.
- MicroRNA-21 (miR-21) is an oncogenic miRNA implicated in tumor cell drug resistance.
Purpose of the Study:
- To investigate the role of miR-21 in human glioma cells resistant to carmustine (BCNU).
- To elucidate the molecular mechanism by which miR-21 influences BCNU resistance.
Main Methods:
- Real-time fluorescence quantitative PCR to measure miR-21 expression.
- Cell Counting Kit-8 (CCK-8) assay to determine BCNU drug sensitivity.
- Western blotting to assess Spry2 protein levels.
Main Results:
- miR-21 expression was significantly higher in BCNU-resistant glioma cells (SWOZ2-BCNU) compared to sensitive cells (SWOZ2).
- BCNU-resistant cells exhibited higher IC50 values for BCNU.
- Reduced Spry2 protein levels were observed in BCNU-resistant cells and correlated inversely with miR-21 expression.
- Silencing Spry2 confirmed its role in mediating miR-21's effect on BCNU resistance.
Conclusions:
- miR-21 enhances BCNU resistance in human glioma cells by downregulating Spry2 protein expression.
- Spry2 represents a potential therapeutic target for overcoming BCNU resistance in gliomas.
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