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The objective of present experiment was to reveal influence of taurine, nanoparticles of gold or nanoparticles of gold conjugated with taurine on molecular and morphological status of muscle’s fibre proliferation activity at the end of embryogenesis. The expression of Proliferating Cell Nuclear Antigen (PCNA) was determined in embryonic chicken pectoral muscle using real time PCR and immunohistochemical method. Furthermore, we calculated the average number of nuclei within the muscle fibre fasciculus in cross sectional image. Taurine, supplemented in ovo to chicken embryo, enhanced organization of breast muscle by activating molecular mechanisms evaluated as an expression of PCNA. Nanoparticles of gold did not influence examined parameters.
Retinoic acid and transforming growth factor-ß (TGF-ß) affect differentiation, pro­liferation and carcinogenesis of epithelial cells. The effect of both compounds on the proliferation of cells of the hormone sensitive human breast cancer cell line (ER+) MCF-7 was assessed in the presence of estradiol and tamoxifen. The assay was based on [ 3H] thymidine incorporation and the proliferative activity of PCNA- and Ki 67-positive cells. The apoptotic index and expression of the Bcl-2 and p53 antigens in MCF-7 cells were also determined. Exogenous TGF-ß1 added to the cell culture showed antiproliferative activity within the concentration range of 0.003-30 ng/ml. Irrespective of TGF-ß 1 concentrations, a marked reduction in the stimulatory action of estradiol (10-9 and 10-8 M) was observed whereas in combination with tamoxifen (10 -7 and 10 -6 M) only 30 ng/ml TGF-ß 1 caused a statistically significant reduction to aproximately 30% of the proliferative cells. In further experiments we examined the effect of exposure of breast cancer cells to retinoids in combination with TGF-ß 1. The incorporation of [3H]thymidine into MCF-7 cells was inhibited to 52 ± 19% (con­trol = 100%) by 3 ng/ml TGF-ß1, and this dose was used throughout. It was found that addition of TGF-ß1 and isotretinoin to the culture did not decrease proliferation, while TGF-ß1 and tretinoin at low concentrations (3 x 10 -8 and 3 x 10-7 M) reduced the percentage of proliferating cells by aproximately 30% (67±8% and 67±5%, P < 0.05 compared to values in the tretinoin group). Both retinoids also led to a sta­tistically significant decrease in the stimulatory effect of 10-9 M estradiol, attenu­ated by TGF-ß1. In addition, the retinoids in combination with TGF-ß1 and tamoxifen (10–6 M) caused a further reduction in the percentage of proliferating cells. Immunocytochemical analysis showed that all the examined compounds gave a statistically significant reduction in the percentage of cells with a positive reaction to PCNA and Ki 67 antigen. TGF-ß1, isotretinoin and tretinoin added to the culture resulted in the lowest percentage of PCNA positive cells. However, the lowest fraction of Ki 67 positive cells was observed after addition of isotretinoin. The obtained results also confirm the fact that the well-known regulatory proteins Bcl-2 and p53 play an important role in the regulation of apoptosis in the MCF-7 cell line, with lowered Bcl-2 expression accompanying easier apoptotic induction. The majority of the examined compounds act via the p53 pathway although some bypass this important proapoptotic factor.
Enoant® is a nutritional reinforcement produced from the grape’s stem, peel and seeds. In recent years grape products such as wine and grape juice have acquired a great importance because of their polyphenol components, which have a strong antioxidant effect. Those antioxidant effects of polyphenols have an ability to inhibit proliferation and induce apoptosis in cancer depending on the types of cancer and application doses. In our study we have aimed to investigate the tumor regressive effects of Enoant® in solid EAC tumor model in Balb-c mice. Animals were randomly divided into 2 groups in this study. 0.5 ml of Enoant® was administered daily to ENTgroup and the same volume of NaCl 0.9% was administered to the control group. Animals continued to receive those applications till sacrification day. On the 8th day 2 × 10⁶ EAC cells in 0.5 ml NaCl 0.9% were injected subcutaneously into the mice’s napes. On day 22 all animals were sacrificed under ether anesthesia. Using PCNA immunohistochemical staining, TUNEL technique, we observed the proliferative and apoptotic cell density changes in tumor tissues as well as the effect of Enoant® on these two phenomena. Dietary Enoant® significantly regressed tumor development in mice. It has been observed that the administration of Enoant® displayed positive effects on EAC tumor’s weight and size when compared with control group animals. Mean tumor weights’ meaningfulness was p < 0.01 and mean short-long diameters’ meaningfulness were p < 0.05 and p<0.01, respectively. It has been determined that while the PCNA index was low (p < 0.05) in the Enoant® administered group, the apoptotic index that has been established with TUNEL technique was high (p < 0.01). As a result, Enoant® has a regressive function on EAC tumor cells. By inducing apoptosis, ENT inhibited the development of tumors. It is thought that ability of ENT was welded from its strong antioxidant polyphenol component. Because of that the use of Enoant® as a dietary supplement is thought to be a factor for inhibiting cancer development.
The aim of the study was to assess the prognostic value of cell proliferation for the continuing course of the disease after the surgical excision of malignant mammary tumours in female dogs. The expression of the proliferating cell nuclear antigen (PCNA) and Ki-67 antigen has been determined by means of immunohistochemistry in formalin-fixed, paraffin-embedded tumour tissue samples taken during the mastectomy. Differences in the intensity of cell nuclei staining have been observed between the PCNA and Ki-67 antigens. The PCNA index was generally twice as high as that of Ki-67; however, the Ki-67 antigen index achieved higher values in predicting the remission time and survival time than the PCNA index. At the same time, only the mean value of the Ki-67 antigen index was statistically higher in the group of dogs with disease progression, than in the groups of dogs without recurrence and metastases. The PCNA index values demonstrated a positive correlation with the degree of histologic malignancy. The results confirm that the determination of the Ki-67 antigen index has a higher prognostic value, especially in the context of its introduction into a standard histopathological examination.
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