Browsing by Subject "PCNA"
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- PublicationOpen AccessExpression of p53 and selected proliferative markers (Ki-67, MCM3, PCNA, and topoisomerase IIα) in borderline ovarian tumors: Correlation with clinicopathological features(Universidad de Murcia. Departamento de Biología Celular e Histología, 2018) Ciepliński, Klaudiusz; Jóźwik, Maciej; Semczuk Sikora, Anna; Gogacz, Marek; Lewkowicz, Dorota; Ignatov, Atanas; Semczuk, AndrzejBackground. The expression of p53 has been studied not only in primary human ovarian carcinomas, but also in borderline ovarian tumors, however, the results were discordant. Expression patterns of proteins involved in cell proliferation and apoptosis have been investigated in various human neoplasms, including female genital tract neoplasms. Objective. The aim of this investigation was to assess the staining pattern and immunolocalization of p53 and selected proliferative markers (Ki-67, MCM3, PCNA, and topoisomerase IIα) in borderline ovarian tumors (BOTs). Design. The study group consisted of 42 women who underwent pelvic surgery between 2006-2015. The median patients’ age was 46 years. The immunoperoxidase technique was employed using antibodies against p53, Ki-67, MCM3, PCNA, and topoisomerase IIα. Results. For p53, nuclear expression was observed in BOTs, however, cytoplasmatic immunoreactivity was also detected. Altogether, 25 (60%) tumors demonstrated positive p53 immunostaining, including overexpression found in 6 (14%). There were no significant differences in p53 expression between subgroups of clinicopathological variables. Immunoexpression of Ki-67, MCM3, PCNA, and topoisomerase IIα was nuclear. Ki67 expression was positive in 12 (29%) cases and there was a trend towards a relationship between patients’ age and Ki-67 staining (P=0.08). Interestingly, a significantly higher Ki-67 expression was found in tumors of ≥10 cm in diameter compared to smaller tumors (P=0.008). MCM3 expression was detected in 38 (90%) tumors, and PCNA expression in 28 (67%), yet none of clinico-pathological factors was related to them. Topoisomerase IIα expression was present in 14 (33%) cases and, interestingly, its significantly higher expression was observed in BOTs of ≥10 cm in diameter compared to smaller tumors (P=0.008). Moreover, Spearman’s correlation revealed highly significant positive associations between Ki-67 and topoisomerase IIα (R=0.403, P=0.008) and Ki-67 and MCM3 (R=0.469, P=0.001). Conclusions. We report a high positive immunostaining rate for p53, suggesting a role of TP53 alterations in the development of BOTs in humans. The new finding of higher topoisomerase IIα immunostaining positivity in BOTs of ≥10 cm may be clinically relevant and requires further studies on larger patient groups.
- PublicationOpen AccessMaternal exposure to the environmental pollutant "BDE-47" impairs the postnatal development of rat cerebellar cortex by modulating neuronal proliferation, synaptogenesis, NGF and BDNF pathways(Universidad de Murcia, Departamento de Biologia Celular e Histiologia, 2022) Mandour, Dalia A.; Tolba, Asmaa M; El Bestawy, Emtethal M.. 2,2’,4,4’-Tetrabromodiphenyl ether (BDE47) is an environmental contaminant that crosses the blood placental barrier and interferes with the homeostasis of fetal thyroid hormones. Aim of work. This study was designed to investigate the perinatal effect of BDE-47 exposure on the postnatal development of the rat cerebellar cortex. Materials and methods. This study was carried out on 20 pregnant rats and 36 of their offspring. The pregnant rats were divided equally into control and BDE-47 treated mother groups; supplemented orally with BDE-47 (0.2 mg/kg/day from day 8 of gestation until the day of weaning). The offspring of both mother groups were subdivided, according to their developmental age, into three subgroups; PND14, PND21and PND42. SerumT3, T4 and TSH were assessed for dams and their offspring. Testing the motor coordination of the offspring via the rotarod test was conducted. Sections of the cerebellar cortex from offspring subgroups were stained with hematoxylin and eosin alongside immunohistochemical reactions and optical density of nerve growth factor (NGF), brain derived neurotrophic factor (BDNF), proliferating cell nuclear antigen (PCNA) and synaptophysin (SYN) were assessed. Also, the thickness of different layers of the cerebellar cortex was histomorphometrically measured. Results. BDE-47 treated mothers and their offspring subgroups showed a significant decrease in the serum free T3, T4 and increased TSH. The BDE-47 offspring displayed incoordination of the motor activity together with disturbed cytoarchitecture of the cerebellar cortical layers, and impaired migration of its germinative neuronal zones, particularly on PND14 and PND21. Moreover, these offspring displayed a decrease of the immune-expression and optical density of NGF, BDNF in the cerebellar cortical layers with impaired proliferation, and synaptogenesis. Conclusion. Maternal exposure to BDE-47 during pregnancy and lactation effectuated a potential deleterious retarding effect on the postnatal development of the rat cerebellar cortex mostly via modulating neuronal proliferation, synaptogenesis, NGF and BDNF pathways secondary to its hypothyroid effect.
- PublicationOpen AccessPeanut lectin-binding sites and mucins in benign and malignant colorectal tissues associated with schistomatosis(Murcia : F. Hernández, 1998) Lin, M.; Hanai, J.; Gui, L.An immunohistochemical and histochemical comparative study was carried out in benign and malignant colorectal tissues with and without schistosomiasis. This included a quantitative determination of peanut lectin (PNA)-binding sites and proliferating cell nuclear antigen (PCNA) expression and histochemical detection of mucin changes. 133 cases were studied, including 70 cases of colorectal carcinoma associated with schistosomiasis (CCS) and 63 cases of colorectal carcinoma without schistosomiasis (CC). Significant differences were found in the type of mucin-containing carcinomas (MC) between CCS and CC. 65% of nontumorous mucosa adjacent to MC of the CCS group expressed PNA-binding sites, significantly higher than those of the MC in the CC group (31%). The nontumorous mucosa in cases of MC of the CCS group also showed a high percentage of sialomucin-predominant secretion (69%, vs 38% in MC of the CC group). Consistently, the presence of PNA-binding sites in MC tumors of the CCS group was increased, compared with that in the same subtype in the CC group (respectively 65% and 31% of strong positivity for PNA). However, no differences in expression of PNA and mucin changes were demonstrated in the surrounding mucosa and tumorous tissues of non-mucin-containing carcinomas (NMC) between CCS and CC. The expression of PCNA was not different between CCS and CC and their subtypes. Our findings suggest a close relationship between mucin-containing colorectal carcinomas and schistosomiasis japonica.
- PublicationOpen AccessQuantitative cell-cycle protein expression in oral cancer assessed by computer-assisted system(Murcia : F. Hernández, 2006) Soares, C.P.; Zuanon, J.A.S.; Teresa, D.B.; Fregonezi, P.A.; Neto, C.B.; Oliveira, M.R.B.; Donadi, E.A.; Martinelli-Kläy, C.P.; Soares, E.G.The knowledge of cell-cycle control has shown that the capacity of malignant growth is acquired by the stepwise accumulation of defects in specific genes regulating cell growth. Histologic diagnosis might be improved by a quantitative evaluation of more specific diagnosis biomarkers, which could help to precisely identify pre-malignant and malignant oral lesions. The aim of the present study is to evaluate whether computer-based quantitative assessment of p53, PCNA and Ki-67 immunohistochemical expression, could be used clinically to foresee the risk of oral malignant transformation. This retrospective study was carried out in ninety-five oral biopsies, 27 were classified as fibrous inflammatory hyperplasia, 40 as leukoplakia and 28 as oral squamous cell carcinoma. Sixteen out of the 40 leukoplakia were diagnosed as non-dysplastic leukoplakia, the other 24 being dysplastic leukoplakia, of which 50.0% were classified as moderate to severe dysplasia. Comparison of the four groups of oral tissues showed significant rises in p53 and Ki-67 positivity index, which increased steadily in the order benign, premalignant, and malignant. In contrast, it was not possible to relate higher PCNA levels with pre-malignant and malignant oral lesions. We therefore conclude that PCNA immunohistochemistry expression is probably an inappropriate marker to identify oral carcinogenesis, whereas joint quantitative evaluation of p53 and Ki-67, appears to be useful as a tumor marker, providing a prediagnostic estimate of the potential for cell-cycle deregulation of the oral proliferate status.
- PublicationOpen AccessSunburn reaction in the dorsal skin of hypotrichotic WBN/ILA-Ht rats(Murcia : F. Hernández, 2003) Okada, T.; Albarenque, S.M.; Yasoshima, A.; Malcotti, V.; Katayama, K.; Uetsuka, K.; Nakayama, Hiroyuki; Doi, K.The dorsal skin responses to a single irradiation with a high-dose of UVB (10kJ/m2) were examined histologically and immunohistochemically in UVB-sensitive Wistar-derived hypotrichotic WBN/ILAHt rats (HtRs). Sunburn cells (SBCs) which were characterized by pyknotic nuclei and eosinophilic cytoplasm and had ultrastructual characteristics of apoptotic cells were first observed in the epidermis at 3 hours (h) after irradiation. The number peaked at 6 h, and then decreased rapidly. The expressions of p53 protein, which is known to be closely related to the formation of SBCs, and of p21 protein, which is one of the transcriptional target genes of p53, were immunohistochemically detected, and their labeling index (LI) in the epidermis peaked at 12 to 24 h (p53) or at 24h (p21) after irradiation. On the other hand, proliferating cell nuclear antigen (PCNA)-LI in keratinocytes was significantly lower than the control group at 6 h after irradiation and thereafter it increased and became significantly higher than the control group from 24 to 48 h. At 48 h, moderate hyperplasia with moderate numbers of mitotic keratinocytes was first observed in the epidermis. In the dermis, mild edema developed from 12 to 36 h and it accompanied mild lymphocyte infiltration at 36 h. Judging from the present results, it was suggested that some factors other than p53 might be involved in SBC formation, and that p53 might induce p21 protein and play an important role in cell growth arrest in keratinocytes after UVB irradiation.
- PublicationOpen AccessThe influence of immunosuppressants on the morphology, proliferating cell nuclear antigen (PCNA) and apoptosis in the rat ventral prostate(F. Hernández y Juan F. Madrid. Universidad de Murcia: Departamento de Biología Celular e Histología, 2015) Grabowska, Marta; Słuczanowska-Głąbowska, Sylwia; Kram, Andrzej; Teresiński, Leszek; Piasecka, Małgorzat; Podhorska-Okołów, Marzenna; Rotter, Iwona; Kędzierska, Karolina; Sporniak-Tutak, Katarzyna; Ciechanowski, Kazimierz; Laszczyńska, MariaAim: Analysis of the impact of immunosuppressants on apoptosis and PCNA in the rat ventral prostate. Method: The studies were performed on 48 male Wistar rats. The animals were divided into a control group and 7 experimental groups. For 6 months, the rats were administered drugs such as: rapamycin (Rapa), cyclosporin A (CsA), tacrolimus (Tac), mycophenolate mofetil (MMF) and glucocorticosteroids (GS). During section of the rats, prostate ventral lobes were obtained. Morphological evaluation (HE, PAS), TUNEL assay, PCNA expression analysis and quantitative image computer analysis were performed. Results: The highest percentage of apoptosis in epithelial cells was observed in groups which received two combinations of drugs: (V) CsA, MMF, GS and (VII) Tac, MMF, GS. A much lower percentage of apoptotic epithelial cells was found in the groups where the treatment schemes included rapamycin throughout the duration of the study. Interestingly, the conversion of the treatment to rapamycin caused a significant reduction of apoptosis in epithelial cells as well as in proliferation in both epithelial and stromal cells. Conclusions: On one hand, the obtained results may explain the anticancer activity of rapamycin in reducing the proliferation of epithelial cells, and on the other hand the adverse effect of rapamycin in the form of reduced regeneration of these cells. Taking into account the prostate in isolation from other organs/systems, the dosage scheme with Rapa, Tac and GS would appear to be the most favorable, due to the smallest morphological changes.