Related Experiment Video
Updated: Aug 20, 2026
Analysis of T-cell Receptor-Induced Calcium Influx in Primary Murine T-cells by Full Spectrum Flow Cytometry
Published on: December 16, 2022
Variation of T-type calcium channel protein expression affects cell division of cultured tumor cells
Amith Panner1, Leanne L Cribbs, Gina M Zainelli
1Department of Neurological Surgery and The Brain Tumor Research Center, University of California San Francisco, San Francisco, CA 94115, USA. apanner@cc.ucsf.edu
Abstract:
In this study we investigated the T-type calcium channel and its involvement in the cell division of U87MG cultured glioma cells and N1E-115 neuroblastoma cells. Using Western blot analysis, we found that expression of both alpha1G and alpha1H subunits of the T-type calcium channel decreased during conditions associated with a decrease in proliferation as evidenced by increased expression of cyclin D1, a marker for non-proliferating cells. Both serum starvation and application of mibefradil, a selective T-type calcium channel antagonist, resulted in a 50% decrease in the expression of alpha1G and alpha1H and a 700-900% increase in levels of cyclin D1 in U87MG and N1E-115 cells, respectively. Furthermore, overexpression of the alpha1H subunit resulted in a two-fold increase in cell proliferation compared to control cultures or cultures receiving an empty vector. In contrast, blocking expression of the alpha1G subunit using antisense oligonucleotides lead to a 70% decrease in proliferation of U87MG and N1E-115 cells compared to control cultures or cultures receiving a scrambled oligonucleotide. Our findings suggest that proliferation of U87MG glioma cells and N1E-115 is regulated by T-type calcium channel expression.
Insights
T-type calcium channels regulate cell division in glioma and neuroblastoma cells. Blocking these channels, or reducing their subunits, inhibits proliferation, while increasing alpha1H subunits boosts cell division.
Area of Science:
- Neuroscience
- Molecular Biology
- Cell Biology
Background:
- T-type calcium channels are implicated in various cellular functions.
- Their role in glioma and neuroblastoma cell proliferation requires further elucidation.
Purpose of the Study:
- To investigate the involvement of T-type calcium channels in the cell division of U87MG glioma and N1E-115 neuroblastoma cells.
- To determine the impact of alpha1G and alpha1H T-type calcium channel subunits on cell proliferation.
Main Methods:
- Western blot analysis to assess protein expression levels.
- Utilized serum starvation and mibefradil (a T-type calcium channel antagonist) to modulate channel activity.
- Employed gene manipulation techniques including subunit overexpression and antisense oligonucleotides to alter subunit expression.
Main Results:
- Decreased expression of alpha1G and alpha1H subunits correlated with reduced cell proliferation and increased cyclin D1 levels.
- Serum starvation and mibefradil treatment led to significant downregulation of alpha1G/alpha1H and upregulation of cyclin D1.
- Overexpression of alpha1H subunit doubled cell proliferation, while antisense inhibition of alpha1G subunit reduced proliferation by 70%.
Conclusions:
- T-type calcium channel expression, specifically alpha1G and alpha1H subunits, plays a critical role in regulating the proliferation of U87MG glioma and N1E-115 neuroblastoma cells.
- These findings highlight T-type calcium channels as potential therapeutic targets for glioma and neuroblastoma treatment.
