AIM: To evaluate the efficacy of telomerase activity assay and peritoneal lavage cytology (PLC) examination in peritoneal lavage fluid for the prediction of peritoneal metastasis in gastric cancer patients, and to ...AIM: To evaluate the efficacy of telomerase activity assay and peritoneal lavage cytology (PLC) examination in peritoneal lavage fluid for the prediction of peritoneal metastasis in gastric cancer patients, and to explore the relationship between telomerase activity and proliferating cell nuclear antigen expression.METHODS: Telomeric repeated amplification protocol (TRAP)-enzyme-linked immunosorbent assay (ELISA) was performed to measure the telomerase activity in 60 patients with gastric cancer and 50 with peptic ulcer. PLC analysis of the 60 patients with gastric cancer was used for comparison. The proliferating cell nuclear antigen (PCNA) in gastric carcinoma was immunohistochemically examined.RESULTS: The telomerase activity and PLC positive rate in peritoneal lavage fluid from patients with gastric cancer was 41.7% (25/60), and 25.0% (15/60), respectively. The positive rate of telomerase activity was significantly higher than that Qf PLC in the group of pT, (15/16 vs 9/16, P 〈 0.05), P1-3 (13/13 vs 9/13, P 〈 0.05) and diffuse type (22/42 vs 13/42, P 〈 0.05). The patients with positive telomerase activity, peritoneal metastasis, and serosal invasion had significantly higher levels of average PCNA proliferation index (PI), (55.00 ± 6.59 vs 27.43 ± 7.72, 57.26 ±10.18 vs 29.15 ±8.31, and 49.82 ± 6.74 vs 24.65 ±7.33, respectively, P 〈 0.05).CONCLUSION: The TRAP assay for telomerase activity is a useful adjunct for cytologic method in the diagnosis of peritoneal micrometastasis and well related to higher proliferating activity of gastric cancer. The results of this study also suggest a promising future therapeutic strategy for treating peritoneal dissemination based on telomerase inhibition.展开更多
Objective: The aim of our study was to investigate the effect of Cinobufacini injection on the proliferation and cell cycle of human hepatoma HepG-2 cells. Methods: Cell proliferation was assessed by MTT assay, cell...Objective: The aim of our study was to investigate the effect of Cinobufacini injection on the proliferation and cell cycle of human hepatoma HepG-2 cells. Methods: Cell proliferation was assessed by MTT assay, cell cycle distribution was detected by the flow cytometry (FCM). The expression of Cyclin A, CDK2 mRNA levels were examined by RT-PCR. Quantitative colorimetric assay was used to analyze Cyclin NCDK2 activity in HepG-2 cells. Results: Cinobufacini injection significantly inhibited HepG-2 cells proliferation in dose- and time-dependent ways; FCM analysis showed Cinobufacini injection induced cell cycle arrest at S phase; RT-PCR assay showed Cinobufacini injection down-regulated Cyclin A, CDK2 expression at mRNA levels; Quantitative colorimetric assay showed Cinobufacini injection deceased Cyclin NCDK2 activity in HepG-2 cells. Conclusion: Cinobufacini injection can inhibit human hepatoma HepG-2 cells growth, induce cell apoptosis and induce cell cycle arrest at S phase, the mechanism of which might be partly related to the down-regulation of Cyclin A, CDK2 mRNA expression and inhibition of Cyclin A/CDK2 activity.展开更多
Objective: The aim of our study was to investigate the effect of Cinobufacini injection on the proliferat(on and apoptosis of human hepatocarcinoma HepG-2 cells. Methods: Cells proliferation was assessed by MTT ass...Objective: The aim of our study was to investigate the effect of Cinobufacini injection on the proliferat(on and apoptosis of human hepatocarcinoma HepG-2 cells. Methods: Cells proliferation was assessed by MTT assay, cells morphologic was observed by the inverted microscopy, Annexin V/PI stain was used to detect the apoptosis and necrosis of the tumor ceils. The expression of TOPOI mRNA and TOPO Ⅱ mRNAwere examined by RT-PCR. Results: Cinobufacini injection significantly inhibited HepG-2 cells proliferation in dose- and time-dependent ways. After Cinobufacini injection intervention, HepG-2 cells showed typical morphological changes: cells changed from polygon into round, chromatin looseness and karyolysis were observed. The percentages of apoptosis were 88.49%, 76.02%, 61.73% corresponding to the 48 h interference of 0.42 μg/mL, 0.21 μg/mL, 0.105 μg/mL Cinobufacini injection, perspectively. RT-PCR assay showed that Cinobufacini injection down-regulated TOPOI and TOPO Ⅱ expression at mRNA level. Conclusion: Cinobufacini can inhibit human hepatocarcinoma HepG-2 cells growth and induce tumor cells apoptosis, the mechanism of which might partly relate to the down-regulation of TOPOI mRNA and TOPO Ⅰ mRNA induced by Cinobufacini injection.展开更多
Supernatants from human primary thymic epithelial cell culture were collected. The proliferationpromoting effect of the supernatant on mouse thymocyte, as well as on Tcell line 85, Be 13, HDMar, Peer, Loucy, Molt4, R...Supernatants from human primary thymic epithelial cell culture were collected. The proliferationpromoting effect of the supernatant on mouse thymocyte, as well as on Tcell line 85, Be 13, HDMar, Peer, Loucy, Molt4, Reh and Jurkat, were observed. The results demonstrated that the supernatants could increase spontaneous 3 HTdR intake of mouse thymocyte, promote ConAinduced thymocyte proliferation and stimulate the proliferation of 85 cell or the cells of HDMar, Loucy and Jurkat at stationary phase, but did not display any effect on Be13, Peer, Molt4 and Reh cells.展开更多
基金the National Natural Science Foundation of China, No. 30370639
文摘AIM: To evaluate the efficacy of telomerase activity assay and peritoneal lavage cytology (PLC) examination in peritoneal lavage fluid for the prediction of peritoneal metastasis in gastric cancer patients, and to explore the relationship between telomerase activity and proliferating cell nuclear antigen expression.METHODS: Telomeric repeated amplification protocol (TRAP)-enzyme-linked immunosorbent assay (ELISA) was performed to measure the telomerase activity in 60 patients with gastric cancer and 50 with peptic ulcer. PLC analysis of the 60 patients with gastric cancer was used for comparison. The proliferating cell nuclear antigen (PCNA) in gastric carcinoma was immunohistochemically examined.RESULTS: The telomerase activity and PLC positive rate in peritoneal lavage fluid from patients with gastric cancer was 41.7% (25/60), and 25.0% (15/60), respectively. The positive rate of telomerase activity was significantly higher than that Qf PLC in the group of pT, (15/16 vs 9/16, P 〈 0.05), P1-3 (13/13 vs 9/13, P 〈 0.05) and diffuse type (22/42 vs 13/42, P 〈 0.05). The patients with positive telomerase activity, peritoneal metastasis, and serosal invasion had significantly higher levels of average PCNA proliferation index (PI), (55.00 ± 6.59 vs 27.43 ± 7.72, 57.26 ±10.18 vs 29.15 ±8.31, and 49.82 ± 6.74 vs 24.65 ±7.33, respectively, P 〈 0.05).CONCLUSION: The TRAP assay for telomerase activity is a useful adjunct for cytologic method in the diagnosis of peritoneal micrometastasis and well related to higher proliferating activity of gastric cancer. The results of this study also suggest a promising future therapeutic strategy for treating peritoneal dissemination based on telomerase inhibition.
文摘Objective: The aim of our study was to investigate the effect of Cinobufacini injection on the proliferation and cell cycle of human hepatoma HepG-2 cells. Methods: Cell proliferation was assessed by MTT assay, cell cycle distribution was detected by the flow cytometry (FCM). The expression of Cyclin A, CDK2 mRNA levels were examined by RT-PCR. Quantitative colorimetric assay was used to analyze Cyclin NCDK2 activity in HepG-2 cells. Results: Cinobufacini injection significantly inhibited HepG-2 cells proliferation in dose- and time-dependent ways; FCM analysis showed Cinobufacini injection induced cell cycle arrest at S phase; RT-PCR assay showed Cinobufacini injection down-regulated Cyclin A, CDK2 expression at mRNA levels; Quantitative colorimetric assay showed Cinobufacini injection deceased Cyclin NCDK2 activity in HepG-2 cells. Conclusion: Cinobufacini injection can inhibit human hepatoma HepG-2 cells growth, induce cell apoptosis and induce cell cycle arrest at S phase, the mechanism of which might be partly related to the down-regulation of Cyclin A, CDK2 mRNA expression and inhibition of Cyclin A/CDK2 activity.
文摘Objective: The aim of our study was to investigate the effect of Cinobufacini injection on the proliferat(on and apoptosis of human hepatocarcinoma HepG-2 cells. Methods: Cells proliferation was assessed by MTT assay, cells morphologic was observed by the inverted microscopy, Annexin V/PI stain was used to detect the apoptosis and necrosis of the tumor ceils. The expression of TOPOI mRNA and TOPO Ⅱ mRNAwere examined by RT-PCR. Results: Cinobufacini injection significantly inhibited HepG-2 cells proliferation in dose- and time-dependent ways. After Cinobufacini injection intervention, HepG-2 cells showed typical morphological changes: cells changed from polygon into round, chromatin looseness and karyolysis were observed. The percentages of apoptosis were 88.49%, 76.02%, 61.73% corresponding to the 48 h interference of 0.42 μg/mL, 0.21 μg/mL, 0.105 μg/mL Cinobufacini injection, perspectively. RT-PCR assay showed that Cinobufacini injection down-regulated TOPOI and TOPO Ⅱ expression at mRNA level. Conclusion: Cinobufacini can inhibit human hepatocarcinoma HepG-2 cells growth and induce tumor cells apoptosis, the mechanism of which might partly relate to the down-regulation of TOPOI mRNA and TOPO Ⅰ mRNA induced by Cinobufacini injection.
文摘Supernatants from human primary thymic epithelial cell culture were collected. The proliferationpromoting effect of the supernatant on mouse thymocyte, as well as on Tcell line 85, Be 13, HDMar, Peer, Loucy, Molt4, Reh and Jurkat, were observed. The results demonstrated that the supernatants could increase spontaneous 3 HTdR intake of mouse thymocyte, promote ConAinduced thymocyte proliferation and stimulate the proliferation of 85 cell or the cells of HDMar, Loucy and Jurkat at stationary phase, but did not display any effect on Be13, Peer, Molt4 and Reh cells.