AI Article Synopsis
- The study investigates the impact of warm ischemia time (WIT) on the expression of HER2, a protein important in breast cancer diagnostics, hypothesizing that hypoxia during WIT may increase HER2 IHC scores.
- Researchers measured HER2 mRNA levels in breast cancer cell lines and found that hypoxia increased the presence of HER2 on the cell membrane but did not affect its mRNA levels.
- The conclusion emphasizes that while HER2 mRNA isn’t upregulated by hypoxia, the amount of HER2 protein on the membrane surfaces does increase, suggesting an important distinction in how HER2 is expressed in hypoxic conditions.
Article Abstract
Background/aim: Tissue specimen quality is becoming increasingly important for basic research and routine clinical results. Warm ischemia time (WIT) affects human epidermal growth factor receptor 2 (HER2) immunohistochemistry (IHC) scores. However, the role of WIT on HER2 modulation remains unclear. We hypothesized that the WIT-mediated increase in HER2 IHC scores was caused by hypoxia. Therefore, this study aimed to determine the mechanism by which WIT mediates the increase in HER2.
Materials And Methods: HER2 mRNA expression was measured in 4T1, SKBR3, and HCC1954 breast cancer cell lines using real-time PCR following hypoxia exposure. The membrane proteins were isolated and extracted using the Mem-PER™ Plus Membrane Protein Extraction Kit (Thermo Fisher Scientific, Waltham, MA, USA) or evaluated through non-permeabilized immunofluorescent analysis.
Results: Hypoxic conditions up-regulated GLUT1 mRNA expression but not HER2 expression. The HER2 membrane protein fraction increased in response to hypoxic conditions. Nonpermeabilized immunofluorescence analysis showed that membrane-bound HER2 was also promoted under hypoxic conditions.
Conclusion: HER2 is not regulated at the mRNA level; however, the level of membrane-bound HER2 increases in response to hypoxia.
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| http://dx.doi.org/10.21873/anticanres.17344 | DOI Listing |
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