Title of article :
Minimal HER1 and HER2 expressions in CHO and HEK-293 cells cause them appropriate negative cells for HERs-related studies
Author/Authors :
Hosseinzadeh Colagar، Abasalt نويسنده Department of Molecular and Cell Biology, Faculty of Basic Sciences, University of Mazandaran, Babolsar, Iran , , Amjadi، Omolbanin نويسنده Department of Molecular and Cell Biology, Faculty of Basic Sciences, University of Mazandaran, Babolsar, Iran , , Rafiei، Alireza نويسنده ,
Issue Information :
فصلنامه با شماره پیاپی سال 2013
Abstract :
Background and Aim: Understanding of cellular and molecular mechanisms
involved in cell/tumor growth and progression has led to molecular-targeted
therapy. HER1 and HER2 are two main oncogenic components of HER-receptor
family considered as targets for cancer research and therapy. Reliable results of
HER- related research and targeted therapies dependent upon cell line with known
HER expression status. But there are no published studies focusing on
determination of HER1 and HER2 receptors expression in CHO and HEK-293 cell
lines.
Methods: Absolute expressions of HER1 and HER2 in CHO, HEK-293, MDAMB-
468 and SKBR3 cells were evaluated using quantitative real-time PCR.
Absolute expression quantification was carried out by serial dilution plasmids
harboring HER1 and HER2 to extrapolate standard curve.
Results: Real-time PCR amplification was successfully optimized by HER1 and
HER2 plasmids with high efficiency and their copy numbers were calculated. The
results showed that HER1 was not expressed in CHO and HEK-293 cells but these
cells express HER2 up to 9.4 × 102 and 1.1 × 105 copy/?g, respectively. MDAMB-
468 and SKBR3 cells express HER1 up to 8.4 × 105 and 9.7 × 104,
respectively. In addition, the expression of HER2 is higher on the surface of
SKBR3 cell (6.5 × 106 copy/?g).
Conclusion: Our study provides a proven insight to HER1 and HER2 expression
status in CHO, HEK-293, MDA-MB-468 and SKBR3 cells. Heterologous
expression of HER1 and HER2 in cell lines is the first step in structural and
functional studies. Since HER1 is not expressed in CHO and HEK-293
endogenously, these cells are ideal for HER receptor related studies.
Journal title :
Research in Molecular Medicine
Journal title :
Research in Molecular Medicine