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Intratubular germ cell neoplasia of the human testis: heterogeneous protein expression and relation to invasive potential.

Mitchell RT, E Camacho-Moll M, Macdonald J, Anderson RA, Kelnar CJ, O'Donnell M, Sharpe RM, Smith LB, Grigor KM, Wallace WH, Stoop H, Wolffenbuttel KP, Donat R, Saunders PT, Looijenga LH - Mod. Pathol. (2014)

Bottom Line: We compared these subpopulations with normal human fetal testis and with seminoma cells.OCT4(+)/MAGEA4(-) cells showed a significantly increased rate of proliferation compared with the OCT4(+)/MAGEA4(+) population (12.8 versus 3.4%, P<0.0001) irrespective of histological tumor type, reflected in the predominance of OCT4(+)/MAGEA4(-) cells in the invasive tumor component.Surprisingly, OCT4(+)/MAGEA4(-) cells in patients with preinvasive disease showed significantly higher proliferation compared to those with seminoma or non-seminoma (18.1 versus 10.2 versus 7.2%, P<0.05, respectively).

View Article: PubMed Central - PubMed

Affiliation: 1] MRC Centre for Reproductive Health, The Queen's Medical Research Institute, The University of Edinburgh, Edinburgh, UK [2] Edinburgh Royal Hospital for Sick Children, Edinburgh, UK.

ABSTRACT
Testicular germ cell cancer develops from premalignant intratubular germ cell neoplasia, unclassified cells that are believed to arise from failure of normal maturation of fetal germ cells from gonocytes (OCT4(+)/MAGEA4(-)) into pre-spermatogonia (OCT4(-)/MAGEA4(+)). Intratubular germ cell neoplasia cell subpopulations based on stage of germ cell differentiation have been described, however the importance of these subpopulations in terms of invasive potential has not been reported. We hypothesized that cells expressing an immature (OCT4(+)/MAGEA4(-)) germ cell profile would exhibit an increased proliferation rate compared with those with a mature profile (OCT4(+)/MAGEA4(+)). Therefore, we performed triple immunofluorescence and stereology to quantify the different intratubular germ cell neoplasia cell subpopulations, based on expression of germ cell (OCT4, PLAP, AP2γ, MAGEA4, VASA) and proliferation (Ki67) markers, in testis sections from patients with preinvasive disease, seminoma, and non-seminoma. We compared these subpopulations with normal human fetal testis and with seminoma cells. Heterogeneity of protein expression was demonstrated in intratubular germ cell neoplasia cells with respect to gonocyte and spermatogonial markers. It included an embryonic/fetal germ cell subpopulation lacking expression of the definitive intratubular germ cell neoplasia marker OCT4, that did not correspond to a physiological (fetal) germ cell subpopulation. OCT4(+)/MAGEA4(-) cells showed a significantly increased rate of proliferation compared with the OCT4(+)/MAGEA4(+) population (12.8 versus 3.4%, P<0.0001) irrespective of histological tumor type, reflected in the predominance of OCT4(+)/MAGEA4(-) cells in the invasive tumor component. Surprisingly, OCT4(+)/MAGEA4(-) cells in patients with preinvasive disease showed significantly higher proliferation compared to those with seminoma or non-seminoma (18.1 versus 10.2 versus 7.2%, P<0.05, respectively). In conclusion, this study has demonstrated that OCT4(+)/MAGEA4(-) cells are the most frequent and most proliferative cell population in tubules containing intratubular germ cell neoplasia, which appears to be an important factor in determining invasive potential of intratubular germ cell neoplasia to seminomas.

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Proliferation of intratubular germ cell neoplasia cells based on diagnosis of pre-invasive disease (PRE INV; n=7), seminoma (SEM; n=7) or non-seminoma (NON-SEM; n=8). A) Proliferation of OCT4+ intratubular germ cell neoplasia cells. B) Proliferation of OCT4+/MAGEA4+ intratubular germ cell neoplasia cells. Mean +/− SEM. * p<0.05, ** p<0.01, in comparison with pre-invasive intratubular germ cell neoplasia.
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Figure 7: Proliferation of intratubular germ cell neoplasia cells based on diagnosis of pre-invasive disease (PRE INV; n=7), seminoma (SEM; n=7) or non-seminoma (NON-SEM; n=8). A) Proliferation of OCT4+ intratubular germ cell neoplasia cells. B) Proliferation of OCT4+/MAGEA4+ intratubular germ cell neoplasia cells. Mean +/− SEM. * p<0.05, ** p<0.01, in comparison with pre-invasive intratubular germ cell neoplasia.

Mentions: In order to investigate whether the germ cell expression profile of the putative intratubular germ cell neoplasia cells might determine their proliferation rate, which might in turn affect their invasive potential, triple immunofluorescence for OCT4/MAGEA4/Ki67 was performed (Supp. Fig. 4). Overall, the proportion of proliferating (Ki67+/OCT4+) intratubular germ cell neoplasia cells was 8.1% (Fig. 6A). There was a shift towards increased proliferation in intratubular germ cell neoplasia cells from patients with pre-invasive disease compared with those with a seminoma or non-seminoma, but this was not statistically significant (Supp. Fig. 5). However, when intratubular germ cell neoplasia (identified by expression of OCT4+) cell proliferation was analysed according to whether or not the cells also expressed MAGEA4, a significantly higher proliferation rate was found for the OCT4+/MAGEA4− population compared to OCT4+/MAGEA4+ intratubular germ cell neoplasia cells (12.8 v 3.4%, p<0.0001; Fig. 6B). Moreover, the significant difference in proliferation rate between the two intratubular germ cell neoplasia phenotypes was consistent when the same analysis was performed according to whether the intratubular germ cell neoplasia cells were from patients with pre-invasive disease (child or adult), seminoma or non-seminoma (Fig. 6C-F). Furthermore, when the proliferation rates in the two sub-populations of intratubular germ cell neoplasia cells (OCT4+/MAGEA4− or OCT4+/MAGEA4+) were compared according to the histology of the adjacent testicular germ cell cancer, there was a significantly higher proliferation rate in the OCT4+/MAGEA4− cells in pre-invasive disease compared with these cells in seminoma and non-seminoma (Fig. 7A); in contrast, there was no difference in the proportion of OCT4+/MAGEA4+ cells that were proliferating for the different tumour types (Fig. 7B).


Intratubular germ cell neoplasia of the human testis: heterogeneous protein expression and relation to invasive potential.

Mitchell RT, E Camacho-Moll M, Macdonald J, Anderson RA, Kelnar CJ, O'Donnell M, Sharpe RM, Smith LB, Grigor KM, Wallace WH, Stoop H, Wolffenbuttel KP, Donat R, Saunders PT, Looijenga LH - Mod. Pathol. (2014)

Proliferation of intratubular germ cell neoplasia cells based on diagnosis of pre-invasive disease (PRE INV; n=7), seminoma (SEM; n=7) or non-seminoma (NON-SEM; n=8). A) Proliferation of OCT4+ intratubular germ cell neoplasia cells. B) Proliferation of OCT4+/MAGEA4+ intratubular germ cell neoplasia cells. Mean +/− SEM. * p<0.05, ** p<0.01, in comparison with pre-invasive intratubular germ cell neoplasia.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4012991&req=5

Figure 7: Proliferation of intratubular germ cell neoplasia cells based on diagnosis of pre-invasive disease (PRE INV; n=7), seminoma (SEM; n=7) or non-seminoma (NON-SEM; n=8). A) Proliferation of OCT4+ intratubular germ cell neoplasia cells. B) Proliferation of OCT4+/MAGEA4+ intratubular germ cell neoplasia cells. Mean +/− SEM. * p<0.05, ** p<0.01, in comparison with pre-invasive intratubular germ cell neoplasia.
Mentions: In order to investigate whether the germ cell expression profile of the putative intratubular germ cell neoplasia cells might determine their proliferation rate, which might in turn affect their invasive potential, triple immunofluorescence for OCT4/MAGEA4/Ki67 was performed (Supp. Fig. 4). Overall, the proportion of proliferating (Ki67+/OCT4+) intratubular germ cell neoplasia cells was 8.1% (Fig. 6A). There was a shift towards increased proliferation in intratubular germ cell neoplasia cells from patients with pre-invasive disease compared with those with a seminoma or non-seminoma, but this was not statistically significant (Supp. Fig. 5). However, when intratubular germ cell neoplasia (identified by expression of OCT4+) cell proliferation was analysed according to whether or not the cells also expressed MAGEA4, a significantly higher proliferation rate was found for the OCT4+/MAGEA4− population compared to OCT4+/MAGEA4+ intratubular germ cell neoplasia cells (12.8 v 3.4%, p<0.0001; Fig. 6B). Moreover, the significant difference in proliferation rate between the two intratubular germ cell neoplasia phenotypes was consistent when the same analysis was performed according to whether the intratubular germ cell neoplasia cells were from patients with pre-invasive disease (child or adult), seminoma or non-seminoma (Fig. 6C-F). Furthermore, when the proliferation rates in the two sub-populations of intratubular germ cell neoplasia cells (OCT4+/MAGEA4− or OCT4+/MAGEA4+) were compared according to the histology of the adjacent testicular germ cell cancer, there was a significantly higher proliferation rate in the OCT4+/MAGEA4− cells in pre-invasive disease compared with these cells in seminoma and non-seminoma (Fig. 7A); in contrast, there was no difference in the proportion of OCT4+/MAGEA4+ cells that were proliferating for the different tumour types (Fig. 7B).

Bottom Line: We compared these subpopulations with normal human fetal testis and with seminoma cells.OCT4(+)/MAGEA4(-) cells showed a significantly increased rate of proliferation compared with the OCT4(+)/MAGEA4(+) population (12.8 versus 3.4%, P<0.0001) irrespective of histological tumor type, reflected in the predominance of OCT4(+)/MAGEA4(-) cells in the invasive tumor component.Surprisingly, OCT4(+)/MAGEA4(-) cells in patients with preinvasive disease showed significantly higher proliferation compared to those with seminoma or non-seminoma (18.1 versus 10.2 versus 7.2%, P<0.05, respectively).

View Article: PubMed Central - PubMed

Affiliation: 1] MRC Centre for Reproductive Health, The Queen's Medical Research Institute, The University of Edinburgh, Edinburgh, UK [2] Edinburgh Royal Hospital for Sick Children, Edinburgh, UK.

ABSTRACT
Testicular germ cell cancer develops from premalignant intratubular germ cell neoplasia, unclassified cells that are believed to arise from failure of normal maturation of fetal germ cells from gonocytes (OCT4(+)/MAGEA4(-)) into pre-spermatogonia (OCT4(-)/MAGEA4(+)). Intratubular germ cell neoplasia cell subpopulations based on stage of germ cell differentiation have been described, however the importance of these subpopulations in terms of invasive potential has not been reported. We hypothesized that cells expressing an immature (OCT4(+)/MAGEA4(-)) germ cell profile would exhibit an increased proliferation rate compared with those with a mature profile (OCT4(+)/MAGEA4(+)). Therefore, we performed triple immunofluorescence and stereology to quantify the different intratubular germ cell neoplasia cell subpopulations, based on expression of germ cell (OCT4, PLAP, AP2γ, MAGEA4, VASA) and proliferation (Ki67) markers, in testis sections from patients with preinvasive disease, seminoma, and non-seminoma. We compared these subpopulations with normal human fetal testis and with seminoma cells. Heterogeneity of protein expression was demonstrated in intratubular germ cell neoplasia cells with respect to gonocyte and spermatogonial markers. It included an embryonic/fetal germ cell subpopulation lacking expression of the definitive intratubular germ cell neoplasia marker OCT4, that did not correspond to a physiological (fetal) germ cell subpopulation. OCT4(+)/MAGEA4(-) cells showed a significantly increased rate of proliferation compared with the OCT4(+)/MAGEA4(+) population (12.8 versus 3.4%, P<0.0001) irrespective of histological tumor type, reflected in the predominance of OCT4(+)/MAGEA4(-) cells in the invasive tumor component. Surprisingly, OCT4(+)/MAGEA4(-) cells in patients with preinvasive disease showed significantly higher proliferation compared to those with seminoma or non-seminoma (18.1 versus 10.2 versus 7.2%, P<0.05, respectively). In conclusion, this study has demonstrated that OCT4(+)/MAGEA4(-) cells are the most frequent and most proliferative cell population in tubules containing intratubular germ cell neoplasia, which appears to be an important factor in determining invasive potential of intratubular germ cell neoplasia to seminomas.

Show MeSH
Related in: MedlinePlus