Cell repertoire and proliferation of germinative cells of the model cestode Mesocestoides corti.

The phylum Platyhelminthes shares a unique population of undifferentiated cells responsible for the proliferation capacity needed for cell renewal, growth, tissue repair and regeneration. These cells have been extensively studied in free-living flatworms, whereas in cestodes the presence of a set of undifferentiated cells, known as germinative cells, has been demonstrated in classical morphology studies, but poorly characterized with molecular biology approaches. Furthermore, several genes have been identified as neoblast markers in free-living flatworms that deserve study in cestode models. Here, different cell types of the model cestode Mesocestoides corti were characterized, identifying differentiated and germinative cells. Muscle cells, tegumental cells, calcareous corpuscle precursor cells and excretory system cells were identified, all of which are non-proliferative, differentiated cell types. Besides those, germinative cells were identified as a population of small cells with proliferative capacity in vivo. Primary cell culture experiments in Dulbecco's Modified Eagle Medium (DMEM), Echinococcus hydatid fluid and hepatocyte conditioned media in non-reductive or reductive conditions confirmed that the germinative cells were the only ones with proliferative capacity. Since several genes have been identified as markers of undifferentiated neoblast cells in free-living flatworms, the expression of pumilio and pL10 genes was analysed by qPCR and in situ hybridization, showing that the expression of these genes was stronger in germinative cells but not restricted to this cell type. This study provides the first tools to analyse and further characterise undifferentiated cells in a model cestode.

MYC is expressed in the stromal and epithelial cells of primary breast carcinoma and paired nodal metastases.

The MYC oncogene is directly involved in the proliferation, metabolism, progression and distant metastasis of breast cancer. Since metastatic spread to the lymph nodes is often the first indication of propensity for metastatic dissemination, the MYC status in nodal disease may represent a decision-making variable. However, the analysis of MYC expression in stromal cells, namely cancer-associated fibroblasts (CAFs), which are known to play a critical role in cancer progression, remains poorly reported. The aim of this study was to determine the expression of MYC and other markers, including estrogen receptor (ER), progesterone receptor (PR), human epidermal growth factor receptor 2 (HER2), p53, Ki67, epidermal growth factor receptor (EGFR), phosphorylated AKT (p-AKT) and phospho-mammalian target of rapamycin (p-mTOR) by immunohistochemistry in representative samples from 80 patients with ductal infiltrative breast cancer and 43 paired compromised axillary lymph nodes allocated in tissue microarrays (TMAs). The epithelial and stromal components of primary tumors and respective lymph node metastases were separately analyzed. MYC expression (cytoplasmic and nuclear) was a frequent event in the epithelial and stromal components of the primary tumors. The epithelial cells in the nodal metastases exhibited a trend for decreased MYC expression compared to that in the primary tumors (P=0.08) but retained the original status of the primary tumors for all other markers. The stromal cells were uniformly negative for ER, PR, HER2, p53, Ki67 and EGFR. Comparison of the stromas of primary tumors and respective lymph node metastases revealed a reduced frequency of nuclear MYC in 15% of the cases (P=0.003), whereas p-mTOR followed a similar trend (P=0.09). Analyses of the possible correlations among markers revealed that epithelial nuclear MYC was associated with p53 (P=0.048). This is an original study demonstrating a significant proportion of MYC expression (nuclear or cytoplasmic), as well p-mTOR and p-AKT expression, in the epithelial and stromal components of either the primary tumor or the nodal metastases. CAFs expressing MYC may establish an angiogenic microenvironment supporting cancer survival and facilitating colonization at the nodal metastatic site.

Avaliação da proliferação celular como indicador prognóstico para mastocitomas cutâneos caninos / Evaluation of cellular proliferation as prognostic indicator for canine cutaneous mast cell tumors

Este estudo teve como objetivo avaliar o valor prognóstico de marcadores de proliferação celular em casos de mastocitomas cutâneos caninos. Vinte e três casos foram analisados quanto à expressão imuno-histoquímica de Ki67 e do Antígeno Nuclear de Proliferação Celular (PCNA), sendo subsequentemente acompanhados clinicamente. Observou-se que a expressão de Ki67 mantém relação negativa com a tradicional graduação histopatológica (p= 0,0418; p<0,05 entre os graus I e III), sendo um indicador confiável para o tempo de sobrevida pós-cirúrgica (p=0,0089). A imunoexpressão de PCNA, apesar de estar correlacionada à marcação por Ki67, não apresentou valores estatisticamente significantes na predição da mortalidade em função da doença e do tempo de sobrevida pós-cirúrgico. Os resultados obtidos confirmam que informações sobre a atividade proliferativa tumoral pela detecção imuno-histoquímica de Ki67 podem incrementar a classificação de mastocitomas cutâneos caninos quanto à malignidade.(AU)

This study evaluated the prognostic value of cell proliferation markers for canine cutaneous mast cell tumor cases. Twenty-three cases were analyzed with regard to immuno-histochemical expression of Ki67 and Proliferating Cell Nuclear Antigen (PCNA), and were clinically followed up. Ki67 expression was related to the traditional histopathological grading (p= 0.0418; p<0.05 between grades I and III), and was a reliable indicator of post-surgical survival (p=0.0089). PCNA immunoexpression did not show statistically significant values in the prediction of disease-related mortality and survival, although it is correlated to Ki67 expression. These results confirm that information about tumoral proliferative activity through Ki67 immunohistochemical detection can improve canine cutaneous mast cell tumor grading with regard to malignancy.(AU)

Avaliação da proliferação celular como indicador prognóstico para mastocitomas cutâneos caninos / Evaluation of cellular proliferation as prognostic indicator for canine cutaneous mast cell tumors

Este estudo teve como objetivo avaliar o valor prognóstico de marcadores de proliferação celular em casos de mastocitomas cutâneos caninos. Vinte e três casos foram analisados quanto à expressão imuno-histoquímica de Ki67 e do Antígeno Nuclear de Proliferação Celular (PCNA), sendo subsequentemente acompanhados clinicamente. Observou-se que a expressão de Ki67 mantém relação negativa com a tradicional graduação histopatológica (p= 0,0418; p<0,05 entre os graus I e III), sendo um indicador confiável para o tempo de sobrevida pós-cirúrgica (p=0,0089). A imunoexpressão de PCNA, apesar de estar correlacionada à marcação por Ki67, não apresentou valores estatisticamente significantes na predição da mortalidade em função da doença e do tempo de sobrevida pós-cirúrgico. Os resultados obtidos confirmam que informações sobre a atividade proliferativa tumoral pela detecção imuno-histoquímica de Ki67 podem incrementar a classificação de mastocitomas cutâneos caninos quanto à malignidade.

This study evaluated the prognostic value of cell proliferation markers for canine cutaneous mast cell tumor cases. Twenty-three cases were analyzed with regard to immuno-histochemical expression of Ki67 and Proliferating Cell Nuclear Antigen (PCNA), and were clinically followed up. Ki67 expression was related to the traditional histopathological grading (p= 0.0418; p<0.05 between grades I and III), and was a reliable indicator of post-surgical survival (p=0.0089). PCNA immunoexpression did not show statistically significant values in the prediction of disease-related mortality and survival, although it is correlated to Ki67 expression. These results confirm that information about tumoral proliferative activity through Ki67 immunohistochemical detection can improve canine cutaneous mast cell tumor grading with regard to malignancy.

Diferenciación neuroendocrina y marcadores de proliferación celular en adenocarcinoma de próstata en un grupo de casos con antígeno prostático específico normal vs elevado / Neuroendocrine differentiation and markers of cell proliferation in a group of patients with prostate adenocarcinoma and normal or high serum prostate-specific antigen levels

Objetivo. Investigar la población de células neuroendocrinas y sus características morfológicas en pacientes con cáncer de próstata y antígeno sérico normal versus antígeno sérico elevado. Material y métodos. En 13 años se identificaron 372 casos de cáncer de próstata de los cuales 19 (5.1%) con antígeno sérico normal (Grupo I). Se seleccionaron 16 grupos controles con antígeno sérico elevado y características histopatológicas similares (Grupo II). Se evaluaron porcentaje de necrosis tumoral, invasión vascular y perineural, inmunohistoquímica: sinaptofisina, enolasa neuroespecífica, antígeno prostático específico, Ki-67 y p53. Resultados. En el grupo I, se obtuvieron 61 % de casos positivos para antígeno tisular, 28.6 % sinaptofisina, 7.1 % para enolasa neuroespecífica, 50 % para p53 y 78.6 % para Ki-67. En el grupo II, los resultados fueron: sinaptofisina 13.3%, enolasa-neuroespecífica 26.6%, antígeno tisular 93%, p53 46.6% y Ki-67 66.7%. Con punto de corte de antígeno tisular expresado en < 80% de células neoplásicas, en el grupo I se encontraron 69.2% de casos, y en el grupo II 21.4% (p = 0.02). Conclusiones. El único dato histológico que mostró diferencia significativa fue la expresión tisular de antígeno prostático específico en < 80% de las células neoplásicas en el grupo I. Se asoció el incremento de las células neuroendocrinas con el menor número de células productoras de antígeno tisular; esta situación podría ser más visible al estudiar un mayor número de pacientes con características semejantes.

OBJECTIVE: Study the morphologic characteristics of neuroendocrine cells in prostate cancer with normal versus elevated prostate specific antigen (PSA). MATERIALS AND METHODS: 372 cases of prostate cancer were identified during a 13 year period, of which 19 displayed normal PSA (group I). Sixteen controls with elevated PSA and similar histopathological characteristics (group II) were included. We studied the degree of tumor necrosis, vascular and perineural invasion. Synaptophysin (SP), neuron specific enolase (NSE), PSA, Ki-67 and p53 inmunoreactivity were also analyzed. RESULTS: Group I positive findings were 61% PSA, 28.6% SP. 7.1% NSE, 50%p53, and 78.6% Ki-67. Group II positive findings were 93% PSA, 13.3% SP, 26.6% NSE 46.6% p53, and 66.7% Ki-67. When we used a <80% cut off point for PSA immunoreactivity in tumor cells, 69.2% of group I and 21.4% of group II were found. CONCLUSIONS: The sole histopathological finding that showed statistical significance was the tissular expression of the specific prostatic antigen in 80% of neoplasic cells in group I. The increase of neuroendocrine cells was associated with a smaller number of tissular antigen producing cells, a finding that could be more apparent if we were to study a larger sample size.