Cell proliferation and cell loss in nodule-in-nodule hepatocellular carcinoma.
Ken YamamotoKatsunobu TakenakaKiyoshi KajiyamaMuneaki ShimadaK ShirabeAkinobu TaketomiTetsuya MaedaK Sugimachi
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Abstract:
In order to clarify the significance of the balance between cell proliferation and cell loss during the progression of hepatocellular carcinoma, 16 operative specimens of nodule-in-nodule hepatocellular carcinoma were investigated.In 16 specimens, cell proliferation was evaluated by the expression of Ki-67 nuclear antigen, and cell loss was also examined by the method of terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling (TUNEL). The expressions of p53 protein, bcl-2 protein and Fas antigen were also investigated to clarify the relationship between their expression and cell kinetics.The Ki-67 labeling index of the inner nodules was higher than that for the outer nodules (18.9% vs. 7.2%; p < 0.05) and the TUNEL labeling index of the inner nodules was also higher than that for the outer nodules (12.8% vs. 6.6%; p < 0.05). The increasing rate of the Ki-67 labeling index from Edmondson's grade I to II was 3.9 +/- 3.0, that from grade II to III was 3.9 +/- 2.4, while the increasing rate of the TUNEL labeling index from grade I to II was 2.7 +/- 0.3 and that from grade II to III was 1.7 +/- 0.2 (p < 0.05). p53 Protein was observed in 5 cases, while bcl-2 protein was found in 4 cases in the border area of the inner nodule. However, Fas antigen was found in none of the examined cases. Regarding the Ki-67 positive rate in the inner nodule, the Ki-67 positive rate in the p53 protein positive cases was significantly higher than that in the negative cases (30.3 +/- 15.4 vs. 11.9 +/- 9.2; p < 0.05). However, the TUNEL labeling index was not affected by the expression of those proteins.This study suggested that tumor progression depends on a disturbance in the cell kinetic balance caused not by a decrease in the absolute amount of cell loss but in the chaotic balance between cell loss and cell proliferation.Keywords:
Nodule (geology)
Ki-67
Terminal deoxynucleotidyl transferase
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To explore the effect of Yiqi Jianpi (YQJP, supplementing Qi and invigorating Spleen) recipe on vascular smooth muscle cells (VSMC) apoptosis and it's mechanisms.VSMC apoptosis was detected by terminal deoxynucleotidyl transferase mediated dUTP nicked labeling (TUNEL) analysis, transmission electron microscope (TEM), flow cytometry and DNA agarose electrophoresis. The expression activity of bcl-2, c-myc and p53 genes were determined using Northern blotting.The typical characteristics of apoptotic VSMC was observed following treatment by the YQJP drug serum. The DNA extracted from VSMC revealed a ladder pattern in electrophoresis. Flow cytometric analysis revealed G0/G1 stage cell blocking phenomena, the hypodiploid apoptotic cell increased, displayed typical peak of apoptosis, the rate of cell apoptosis and YQJP serum concentration were in a dose-dependent manner. The result of hybridization showed that 30% of YQJP serum could inhibit apoptotic gene bcl-2 expression, upregulated the level of apoptogenous gene c-myc and p53 mRNA.VSMC apoptosis could be induced by YQJP recipe and its mechanism might be through affecting the apoptosis related gene expression activity.
Agarose gel electrophoresis
Terminal deoxynucleotidyl transferase
Apoptosis Induction
Apoptotic body
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The detergent Triton X-100 was used to establish a model for apoptosis in hepatoma cell lines. The electrophoresis of DNA extracted from 0.01% Triton X-100 treated hepatoma cell lines showed DNA ladder formation, a hallmark of apoptosis. The DNA fragmentation appeared within less than 60 min of the Triton X-100 treatment. Chromatin condensation and apoptotic bodies were observed by hematoxylin and eosin (H & E) stain, and fragmented nucleosome was detected by terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labelling (TUNEL) test. Apoptosis was semi-quantitated by measuring the lactate dehydrogenase (LDH) level for cytotoxity. It was found that apoptosis had been induced in more than 90% of the cells treated with Triton X-100 for 150 min. These data show that Triton X-100 efficiently induces the apoptotic cell death in hepatoma cell lines.
Terminal deoxynucleotidyl transferase
Fragmentation
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Recent molecular studies suggest that the expression of high-risk but not low-risk human papillomavirus (HPV) oncoproteins E6 and E7 can significantly alter normal cell cycle regulation. The alterations in cell cycle regulation may be reflected by changes in the balance between cell growth and cell loss through apoptosis in cell populations expressing E6 and/or E7. We evaluated the kinetic indices of cell proliferation and apoptosis in a histopathological spectrum of cervical neoplasia and compared low-versus high-risk HPV-associated lesions. The cell proliferation index, as determined by detection of the nuclear antigen Ki67, increased with increasing lesion grade. Apoptotic cells were identified with terminal deoxynucleotidyl transferase-labeling of the 3'-hydroxyl ends of DNA nucleosomes. No apoptosis was observed in normal epithelium, and only occasional apoptotic cells were seen in low-grade lesions. However, there was a low but measurable apoptotic index in the higher grade lesions, which increased with lesion grade. There was no significant difference in the proliferative and apoptotic indices in similar grade lesions when stratified into low-versus high-risk HPV types. These findings suggest that apoptosis in HPV-infected lesions correlates with proliferative activity rather than HPV type.
Terminal deoxynucleotidyl transferase
Ki-67
Proliferation index
Papillomaviridae
Proliferative index
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Objective To determine relationship between the expression of Fas antigen and apoptosis in human gastric carcinoma tissues.Method Using immunohistochemical staining and terminal deoxynucleotidyl transferase mediated dUTP nick end labeling(TUNEL) technique,we observed the apoptotic cells and Fas antigen expression in 40 cases of gastric carcinoma tissues.Results The positive rate of Fas antigen expression in gastric carcinoma tissues was 52.5%.The apoptotic index in Fas positive tissues was significantly higher than that in Fas negative tissues( t=3.35,P 0.01).Conclusions The result suggested that cellular apoptosis was closely related to the Fas antigen expression in gastric carcinoma tissues,the abnormal regulation of apoptosis may play an important role in the pathogenesis of gastric carcinoma.
Terminal deoxynucleotidyl transferase
Gastric carcinoma
Fas ligand
Pathogenesis
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AIM To observe the variation of lung cell proliferation and apoptosis before and after LVRS and to investigate the relationship between the lung cell proliferation and apoptosis during chronic anoxia. METHODS Pulmonary emphysema animal models were established and operated a long side LVRS. The animals were sacrificed at 4 wk after LVRS and lungs were removed. The lung cell proliferation and apoptosis were determined quantitatively and qualitatively by immunohistochemical and terminal deoxynucleotidyl transferase mediated dUTP nick end labeling (TUNEL) techniques. RESULTS TUNEL examination showed scattered apoptosis cells with brown nucleus and concentrated or crescent shaped nucleus chromatin under microscope. Proliferated cells displayed Ki 67 nucleus positive reaction. The apoptosis cells and Ki 67 nucleus positive cells were (5.9±1.8)% and (1.7±0.8)% before LVRS, and (3.6±1.2)% and (2.7± 0.9)% after LVRS respectively. CONCLUSION Destruction of the balance between proliferation and apoptosis of the lung cell might result in the increased lung cell apoptosis due to chronic anoxia before LVRS. The lung cell apoptosis and proliferation could gradually reach their normal status following anoxia improvement after LVRS.
Terminal deoxynucleotidyl transferase
Alveolar Wall
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Fas ligand
Terminal deoxynucleotidyl transferase
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Objective To study The relationship between apoptosis and expression of P27 protein in hepatocellular carcinoma(HCC).Methods P27 protein and apoptotic cells were detected by immunohistochemical S-P method and TUNEL method in 86 paraffin-embedded tissues of HCC.Results The positive rate of P27 protein in these tissues was 37.2%(32/86). The mean apoptotic index in the HCC with positive expression of P27 protein (6.72‰) was significantly higher than that in the negative group (2.83‰). Conclusion Expression of P27 protein related significantly with cell apotosis of HCC. Promoting apoptosis may be one of the effective anti-cancer mechanisms of P27 protein.
BAX Protein
Hepatic carcinoma
P53 protein
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Objective To study the toxic effect of glumatic acid on nerve cell of Sprague-Dawley rat by injecting different concentration glumatic acid and to determine the degree of nerve cell apoptosis in different time by using immunohistochemistry technique.Methods Seting SD rat model and observing the toxic effect of glumatic acid on nerve cell at different time by using cross immunity immunohistochemistry technique in situ and TUNEL staining method.Results There was remarkably difference between glumatic acid group and the normal sodium group by counting the number of nerve cell apoptosis(P0.05).Conclusion Specified dosage of glumatic acid can cause nerve cell apoptosis.
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Objective:To study the relation between cell proliferation,apoptosis and the expression of P16 and tumour differentiation in primary hepatocellular carcinoma (HCC). Methods:The study was performed on 50 paraffin fixed hepatic tissue which were randomly collected from HCC patients.By immunohistochemistry methods,Ki 67 and P16 expression were analyzed.Tunel method was adoped to analyze the apoptosis occurrence.x 2 text and correlation analysis were adopted for statistical analysis.Results:Proliferation/apoptosis ratio has a positive correlation to HCC cell differenation,P16 appears in cell nucleus.Conclusion:The positive relation between Ki 67 expression and apoptosis exists.Proliferation and apoptosis can affect HCC in growth and differentiation of HCC.P16 is related to cell proliferation and apoptosis.Inducing apoptosis and inhabit cell proliferation can improve the clinical therapy effects of HCC.
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