In order to investigate the changes of vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF) expression in residual hepatocellular carcinoma (HCC) after transcatheter arterial chemoemboli...In order to investigate the changes of vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF) expression in residual hepatocellular carcinoma (HCC) after transcatheter arterial chemoembolization (TACE), the expression levels of VEGF and bFGF expression in specimens surgically removed from 48 HCC patients were detected by immunohistochemical methods, and staining intensity of VEGF and bFGF was assessed by a computer-assisted image-analyzer Among the 48 patients, 25 underwent partial hepatectomy alone (single operating group), and 23 were subjected to second stage surgical resection after TACE (TACE group) The results showed that the average absorbance value (A) of VEGF was higher in TACE group than that in single operating group (0 152±0 021 vs 0 131±0 012, P< 0 01) The Average A of bFGF in TACE group was 0 127±0 023, higher than in single operating group (0 111±0 016, P <0 05) These results suggested that TACE of HCC can up-regulate the expression of VEGF and bFGF in HCC tissues possibly due to anoxia and ischemia展开更多
AIM: To investigate the inhibitory efficacy of <sup>125</sup>I-labeled anti-basic fibroblast growth factor (bFGF) monoclonal antibody (mAb) in hepatocellular carcinoma (HCC).METHODS: bFGF mAb was prepared ...AIM: To investigate the inhibitory efficacy of <sup>125</sup>I-labeled anti-basic fibroblast growth factor (bFGF) monoclonal antibody (mAb) in hepatocellular carcinoma (HCC).METHODS: bFGF mAb was prepared by using the 1G9B9 hybridoma cell line with hybridization technology and extracted from ascites fluid through a Protein G Sepharose affinity column. After labeling with <sup>125</sup>I through the chloramine-T method, bFGF mAb was further purified by a Sephadex G-25 column. Gamma radiation counter GC-1200 detected radioactivity of <sup>125</sup>I-bFGF mAb. The murine H22 HCC xenograft model was established and randomized to interventions with control (phosphate-buffered saline), <sup>125</sup>I-bFGF mAb, <sup>125</sup>I plus bFGF mAb, bFGF mAb, or <sup>125</sup>I. The ratios of tumor inhibition were then calculated. Expression of bFGF, fibroblast growth factor receptor (FGFR), platelet-derived growth factor, and vascular endothelial growth factor (VEGF) mRNA was determined by quantitative reverse transcriptase real-time polymerase chain reaction.RESULTS: The purified bFGF mAb solution was 8.145 mg/mL with a titer of 1:2560000 and was stored at -20 °C. After coupling, <sup>125</sup>I-bFGF mAb was used at a 1: 1280000 dilution, stored at 4 °C, and its specific radioactivity was 37 MBq/mg. The corresponding tumor weight in the control, <sup>125</sup>I, bFGF mAb, <sup>125</sup>I plus bFGF mAb, and <sup>125</sup>I-bFGF mAb groups was 1.88 ± 0.25, 1.625 ± 0.21, 1.5 ± 0.18, 1.41 ± 0.16, and 0.98 ± 0.11 g, respectively. The tumor inhibition ratio in the <sup>125</sup>I, bFGF mAb, <sup>125</sup>I plus bFGF mAb, and <sup>125</sup>I-bFGF mAb groups was 13.6%, 20.2%, 25.1%, and 47.9%, respectively. Growth of HCC xenografts was inhibited significantly more in the <sup>125</sup>I-bFGF mAb group than in the other groups (P < 0.05). Expression of bFGF and FGFR mRNA in the <sup>125</sup>I-bFGF mAb group was significantly decreased in comparison with other groups (P < 0.05). Groups under interventions revealed increased expression of VEGF mRNA (except for <sup>125</sup>I group) compared with the control group.CONCLUSION: <sup>125</sup>I-bFGF mAb inhibits growth of HCC xenografts. The coupling effect of <sup>125</sup>I-bFGF mAb is more effective than the concomitant use of <sup>125</sup>I and bFGF mAb.展开更多
To observe the effects of basic fibroblast growth factor （bFGF） on human adenoid cystic carcinoma ACC-2 cell line proliferation and ERK, cyclin D1/p21^waf/cip1 signaling pathways, human adenoid cystic carcinoma cel...To observe the effects of basic fibroblast growth factor （bFGF） on human adenoid cystic carcinoma ACC-2 cell line proliferation and ERK, cyclin D1/p21^waf/cip1 signaling pathways, human adenoid cystic carcinoma cells （ACC-2） were cultured and the influence of bFGF of different concentrations on cell proliferation was determined by MTT. Protein was detected by immuno-precipitation and ERK activity by using ERK agent kit. p-ERK1/2 and down-stream cyclin D1, p21^waf/cip1 expression were detected by Western blotting and the interfering role of mitogen protein-activated kinase （MEK） suppressor U0126 in the afore-mentioned indicators was examined. MTT demonstrated ACC-2 cell proliferation was substantially enhanced by bFGF, immuo-precipitation displayed ERK activity was up-regulated by bFGF, and immuno-imprinting also showed p-ERK1/2, cyclin D1 expression was greatly enhanced and p21^waf/cip1 expression was inhibited by bFGE U0126 suppressed the effect of bFGF. It is concluded that bFGF can promote the proliferation of human adenoid cystic carcinoma ACC-2 cells, and its pathways are associated with the up-regulated activity and expression of p-ERK1/2, inhibited p21waf/cip1 expression and enhanced cyclin D1 expression.展开更多
文摘In order to investigate the changes of vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF) expression in residual hepatocellular carcinoma (HCC) after transcatheter arterial chemoembolization (TACE), the expression levels of VEGF and bFGF expression in specimens surgically removed from 48 HCC patients were detected by immunohistochemical methods, and staining intensity of VEGF and bFGF was assessed by a computer-assisted image-analyzer Among the 48 patients, 25 underwent partial hepatectomy alone (single operating group), and 23 were subjected to second stage surgical resection after TACE (TACE group) The results showed that the average absorbance value (A) of VEGF was higher in TACE group than that in single operating group (0 152±0 021 vs 0 131±0 012, P< 0 01) The Average A of bFGF in TACE group was 0 127±0 023, higher than in single operating group (0 111±0 016, P <0 05) These results suggested that TACE of HCC can up-regulate the expression of VEGF and bFGF in HCC tissues possibly due to anoxia and ischemia
基金Supported by the National Natural Science Foundation of China,No.81273814Guangdong Province Key Scientific Research Grant,No.2013A022100031
文摘AIM: To investigate the inhibitory efficacy of <sup>125</sup>I-labeled anti-basic fibroblast growth factor (bFGF) monoclonal antibody (mAb) in hepatocellular carcinoma (HCC).METHODS: bFGF mAb was prepared by using the 1G9B9 hybridoma cell line with hybridization technology and extracted from ascites fluid through a Protein G Sepharose affinity column. After labeling with <sup>125</sup>I through the chloramine-T method, bFGF mAb was further purified by a Sephadex G-25 column. Gamma radiation counter GC-1200 detected radioactivity of <sup>125</sup>I-bFGF mAb. The murine H22 HCC xenograft model was established and randomized to interventions with control (phosphate-buffered saline), <sup>125</sup>I-bFGF mAb, <sup>125</sup>I plus bFGF mAb, bFGF mAb, or <sup>125</sup>I. The ratios of tumor inhibition were then calculated. Expression of bFGF, fibroblast growth factor receptor (FGFR), platelet-derived growth factor, and vascular endothelial growth factor (VEGF) mRNA was determined by quantitative reverse transcriptase real-time polymerase chain reaction.RESULTS: The purified bFGF mAb solution was 8.145 mg/mL with a titer of 1:2560000 and was stored at -20 °C. After coupling, <sup>125</sup>I-bFGF mAb was used at a 1: 1280000 dilution, stored at 4 °C, and its specific radioactivity was 37 MBq/mg. The corresponding tumor weight in the control, <sup>125</sup>I, bFGF mAb, <sup>125</sup>I plus bFGF mAb, and <sup>125</sup>I-bFGF mAb groups was 1.88 ± 0.25, 1.625 ± 0.21, 1.5 ± 0.18, 1.41 ± 0.16, and 0.98 ± 0.11 g, respectively. The tumor inhibition ratio in the <sup>125</sup>I, bFGF mAb, <sup>125</sup>I plus bFGF mAb, and <sup>125</sup>I-bFGF mAb groups was 13.6%, 20.2%, 25.1%, and 47.9%, respectively. Growth of HCC xenografts was inhibited significantly more in the <sup>125</sup>I-bFGF mAb group than in the other groups (P < 0.05). Expression of bFGF and FGFR mRNA in the <sup>125</sup>I-bFGF mAb group was significantly decreased in comparison with other groups (P < 0.05). Groups under interventions revealed increased expression of VEGF mRNA (except for <sup>125</sup>I group) compared with the control group.CONCLUSION: <sup>125</sup>I-bFGF mAb inhibits growth of HCC xenografts. The coupling effect of <sup>125</sup>I-bFGF mAb is more effective than the concomitant use of <sup>125</sup>I and bFGF mAb.
文摘To observe the effects of basic fibroblast growth factor （bFGF） on human adenoid cystic carcinoma ACC-2 cell line proliferation and ERK, cyclin D1/p21^waf/cip1 signaling pathways, human adenoid cystic carcinoma cells （ACC-2） were cultured and the influence of bFGF of different concentrations on cell proliferation was determined by MTT. Protein was detected by immuno-precipitation and ERK activity by using ERK agent kit. p-ERK1/2 and down-stream cyclin D1, p21^waf/cip1 expression were detected by Western blotting and the interfering role of mitogen protein-activated kinase （MEK） suppressor U0126 in the afore-mentioned indicators was examined. MTT demonstrated ACC-2 cell proliferation was substantially enhanced by bFGF, immuo-precipitation displayed ERK activity was up-regulated by bFGF, and immuno-imprinting also showed p-ERK1/2, cyclin D1 expression was greatly enhanced and p21^waf/cip1 expression was inhibited by bFGE U0126 suppressed the effect of bFGF. It is concluded that bFGF can promote the proliferation of human adenoid cystic carcinoma ACC-2 cells, and its pathways are associated with the up-regulated activity and expression of p-ERK1/2, inhibited p21waf/cip1 expression and enhanced cyclin D1 expression.
