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== Immunohistochemical evaluation ==
The immunohistochemical detection of proliferation related proteins such as [[Ki-67 (protein)|Ki-67]] and proliferating cell nuclear antigen is a commonly used method to determine the proliferation index. Ki-67 is a nuclear antigen expressed in proliferating cells that is coded by the MKI67 gene on chromosome 10, and is expressed during the GI, S, G2, and M phases of the cell cycle. Cells are then stained with a Ki-67 antibody, and the number of stained nuclei is then expressed as a percentage of total tumor cells. It is recommended to count at least 500 tumor cells in the highest labeled area. The Ki-67 score closely correlates with other proliferation markers, and has been shown to have prognostic and predictive value for many different tumor types.<ref>{{Cite journal|last=de Azambuja|first=E|last2=Cardoso|first2=F|last3=Castro|first3=G de|last4=Colozza|first4=M|last5=Mano|first5=M S|last6=Durbecq|first6=V|last7=Sotiriou|first7=C|last8=Larsimont|first8=D|last9=Piccart-Gebhart|first9=M J|date=May 2007|title=Ki-67 as prognostic marker in early breast cancer: a meta-analysis of published studies involving 12 155 patients|url=http://www.nature.com/articles/6603756|journal=British Journal of Cancer|language=En|volume=96|issue=10|pages=1504–1513|doi=10.1038/sj.bjc.6603756|issn=1532-1827|pmc=2359936}}</ref> Similarly, [[proliferating cell nuclear antigen]] (PCNA) is a protein associated with cell proliferation that is upregulated in proliferating cells, making it another useful antigen for immunostaining.<ref>{{Cite journal|last=Wang|first=Xing|last2=Wang|first2=Dong|last3=Yuan|first3=Na|last4=Liu|first4=Fanxiao|last5=Wang|first5=Fu|last6=Wang|first6=Bomin|last7=Zhou|first7=Dongsheng|date=2017-10-01|title=The prognostic value of PCNA expression in patients with osteosarcoma|url=http://Insights.ovid.com/crossref?an=00005792-201710130-00023|journal=Medicine|language=ENGLISH|volume=96|issue=41|doi=10.1097/md.0000000000008254|issn=0025-7974}}</ref> It is associated with [[DNA polymerase|DNA polymerase alpha]], which is expressed throughout the phases of the cell cycle. The expression of PCNA also correlates well with other proliferation markers such as mitotic count, S-phase fraction and Ki-67 staining.
== Diagnostic role of proliferation index ==
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