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Nuclear morphometric analysis in tissue as an objective tool with potential use to improve melanoma staging

Nunes, Tatiana W.N.a,,*; Filippi-Chiela, Eduardo C.b,,*; Callegari-Jacques, Sídia M.c; da Silva, Vinicius D.g; Sansonowicz, Tatianad; Lenz, Guidoe; Roehe, Adriana V.f

doi: 10.1097/CMR.0000000000000594
Original Articles: Translational Research
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Alterations in nuclear size and shape are commonly observed in cancers, and its objective evaluation may provide valuable clinical information about the outcome of the disease. Here, we applied the nuclear morphometric analysis in tissues in hematoxylin and eosin-digitized slides of nevi and melanoma, to objectively contribute to the prognostic evaluation of these tumors. To this, we analyzed the nuclear morphometry of 34 melanomas classified according to the TNM stage. Eight cases of melanocytic nevi were used as non-neoplastic tissues to set the non-neoplastic parameters of nuclear morphology. Our samples were set as G1 (control, nevi), G2 (T1T2N0M0), G3 (T3T4N0M0), G4 (T1T2N1M1), and G5 (T3T4N1M1). Image-Pro Plus 6.0 software was used to acquire measurements related to nuclear size (variable: Area) and shape (variables: Aspect, AreaBox, Roundness, and RadiusRatio, which were used to generate the Nuclear Irregularity Index). From these primary variables, a set of secondary variables were generated. All the seven primary and secondary variables related to the nuclear area were different among groups (Pillai’s trace P<0.001), whereas Nuclear Irregularity Index, which is the variable related to nuclear shape, did not differ among groups. The secondary variable ‘Average Area of Large Nuclei’ was able to differ all pairwise comparisons, including thin nonmetastatic from thin metastatic tumors. In conclusion, the objective quantification of nuclear area in hematoxylin and eosin slides may provide objective information about the risk stratification of these tumors and has the potential to be used as an additional method in clinical decision making.

aLaboratory of Pathology of the Hospital Nossa Senhora da Conceição de Porto Alegre

bDepartment of Morphological Sciences, Federal University of Rio Grande do Sul

cDepartment of Statistics, Institute of Mathematics and Statistics of the Federal University of Rio Grande do Sul

dProgram of Medical Residency and Pathology of Hospital Nossa Senhora da Conceição de Porto Alegre

eDepartment of Biophysics and Biotechnology Center, Federal University of Rio Grande do Sul

fDepartment of Pathology, Federal University of Health Sciences of Porto Alegre, Porto Alegre

gDepartment of Pathology, Barretos Cancer Hospital, Barretos, Brazil

* Tatiana W.N. Nunes and Eduardo C. Filippi-Chiela contributed equally to the writing of this article.

Received 13 September 2018 Accepted 24 January 2019

Supplemental Digital Content is available for this article. Direct URL citations appear in the printed text and are provided in the HTML and PDF versions of this article on the journal’s website, www.melanomaresearch.com.

Correspondence to Tatiana W.N. Nunes, PhD, Hospital Conceição, Av. Francisco Trein, 596 - Cristo Redentor, Porto Alegre, Porto Alegre, Rio Grande do Sul 91350-200, Brazil Tel/fax: +55 51 3357 2000; e-mail: tatianawnn@gmail.com

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