Protein and gene expression of estrogen receptor alpha and nuclear morphology of two breast cancer cell lines after different fixation methods.
We assessed morphology and ERalpha protein and gene expression of two breast cancer cell lines after three different fixatives: neutral-buffered 10% formaldehyde, LN-FIX and FineFIX and varying fixation times. We found that the cell morphology was best preserved in cells fixed with LN-FIX. Two commercial fixatives used in this study shrank cells less than formalin. In immunohistochemical assay samples were stained with two different ERalpha antibodies, clone 1D5 and clone SP1. All tested fixatives were suitable for immunohistochemistry. Staining was more intensive and the number of stained cells was larger with the clone 1D5 than with the clone SP1. Our gene expression analysis showed that formalin and LN-FIX preserve the ERalpha better than FineFIX, which is advertised to be optimal for molecular analysis. Our study suggests that tissues fixed with formalin are suitable also for molecular biology assays. This makes possible to research formalin-fixed paraffin-embedded archival tissues also with molecular techniques.
Nykänen
Marjukka
M
Department of Pathology, Central Finland Central Hospital, Keskussairaalantie 19, 40620 Jyväskylä, Finland. marjukka.nykanen@ksshp.fi
Kuopio
Teijo
T
D001943:Breast Neoplasms / Q000235:genetics* / Q000378:metabolism* / Q000473:pathology; D018929:Cell Culture Techniques / Q000379:methods; D045744:Cell Line, Tumor; D002467:Cell Nucleus / Q000235:genetics / Q000378:metabolism*; D047628:Estrogen Receptor alpha / Q000235:genetics* / Q000378:metabolism*; D005260:Female; D005557:Formaldehyde; D015972:Gene Expression Regulation, Neoplastic; D006801:Humans; D007150:Immunohistochemistry / Q000379:methods; D016133:Polymerase Chain Reaction; D012334:RNA, Neoplasm / Q000235:genetics / Q000302:isolation & purification; D048348:Reverse Transcription; D013194:Staining and Labeling / Q000379:methods; D016707:Tissue Fixation / Q000379:methods*
D047628:Estrogen Receptor alpha; D012334:RNA, Neoplasm; D005557:Formaldehyde