The effects of β-NGF controlled delivery system on growth of primary cultured chick embryo dorsal root ganglion axons

Objective To construct β-NGF controlled delivery system and evaluate the biological effects of β-NGF on the growth of chick embryo dorsal root ganglion (DRG) axons in vitro. Methods Delivery systems releasing β-NGF at 50/μL, 100/μg/L and 250 μg/L concentration were constructed. To determine the optimal dose response effects of NGF in the controlled delivery system, DRG were co-cultured with of β-NGF at above concentrations while using DRG basic culture as control. Axonal growth was observed. DRG were also cocultured with the components in the controlled delivery system to detect the effects on growth of DRG axons. The experiment was divided into 5 experimental groups and 1 control group: control group, DRG+ fibrin; Group A,DRG+ fibrin+ peptide + heparin + 100 μg/L β-NGF; Group B, DRG + fibrin + heparin + 100/μg/L β-NGF;Group C, DRG + fibrin + peptide + 100 μg/L β-NGF; Group D, DRG + fibrin + 100 μg/L β-NGF; Group E,DRG + fibrin + peptide + heparin. Results The growth of DRG axons in 50 μg/L, 100μg/L and 250/μg/Lconcentration of β-NGF controlled delivery system was 1.31 ( P > 0. 05), 3.78 ( P < 0. 01 ) and 3.05 ( P <0.01) folds of the control respectively. The growth of DRG axons in 100 μg/L group was significantly better comparing to that in 250 μg/L group. The growth of DRG axons in Groups A, B, C, D and E was 3.75, 1.15,1.12, 1.10 and 1.09 folds of the control group, respectively. The difference was only statistically significant between Group A and the control group ( P < 0. 01 ). Conclusion β-NGF released from the β-NGF controlled delivery system was bioactive. It could promote the growth of DRG axons.     

The effects of β-NGF controlled delivery system on growth of primary cultured chick embryo dorsal root ganglion axons

Objective To construct β-NGF controlled delivery system and evaluate the biological effects of β-NGF on the growth of chick embryo dorsal root ganglion (DRG) axons in vitro. Methods Delivery systems releasing β-NGF at 50/μL, 100/μg/L and 250 μg/L concentration were constructed. To determine the optimal dose response effects of NGF in the controlled delivery system, DRG were co-cultured with of β-NGF at above concentrations while using DRG basic culture as control. Axonal growth was observed. DRG were also cocultured with the components in the controlled delivery system to detect the effects on growth of DRG axons. The experiment was divided into 5 experimental groups and 1 control group: control group, DRG+ fibrin; Group A,DRG+ fibrin+ peptide + heparin + 100 μg/L β-NGF; Group B, DRG + fibrin + heparin + 100/μg/L β-NGF;Group C, DRG + fibrin + peptide + 100 μg/L β-NGF; Group D, DRG + fibrin + 100 μg/L β-NGF; Group E,DRG + fibrin + peptide + heparin. Results The growth of DRG axons in 50 μg/L, 100μg/L and 250/μg/Lconcentration of β-NGF controlled delivery system was 1.31 ( P > 0. 05), 3.78 ( P < 0. 01 ) and 3.05 ( P <0.01) folds of the control respectively. The growth of DRG axons in 100 μg/L group was significantly better comparing to that in 250 μg/L group. The growth of DRG axons in Groups A, B, C, D and E was 3.75, 1.15,1.12, 1.10 and 1.09 folds of the control group, respectively. The difference was only statistically significant between Group A and the control group ( P < 0. 01 ). Conclusion β-NGF released from the β-NGF controlled delivery system was bioactive. It could promote the growth of DRG axons.     

The effects of β-NGF controlled delivery system on growth of primary cultured chick embryo dorsal root ganglion axons

Objective To construct β-NGF controlled delivery system and evaluate the biological effects of β-NGF on the growth of chick embryo dorsal root ganglion (DRG) axons in vitro. Methods Delivery systems releasing β-NGF at 50/μL, 100/μg/L and 250 μg/L concentration were constructed. To determine the optimal dose response effects of NGF in the controlled delivery system, DRG were co-cultured with of β-NGF at above concentrations while using DRG basic culture as control. Axonal growth was observed. DRG were also cocultured with the components in the controlled delivery system to detect the effects on growth of DRG axons. The experiment was divided into 5 experimental groups and 1 control group: control group, DRG+ fibrin; Group A,DRG+ fibrin+ peptide + heparin + 100 μg/L β-NGF; Group B, DRG + fibrin + heparin + 100/μg/L β-NGF;Group C, DRG + fibrin + peptide + 100 μg/L β-NGF; Group D, DRG + fibrin + 100 μg/L β-NGF; Group E,DRG + fibrin + peptide + heparin. Results The growth of DRG axons in 50 μg/L, 100μg/L and 250/μg/Lconcentration of β-NGF controlled delivery system was 1.31 ( P > 0. 05), 3.78 ( P < 0. 01 ) and 3.05 ( P <0.01) folds of the control respectively. The growth of DRG axons in 100 μg/L group was significantly better comparing to that in 250 μg/L group. The growth of DRG axons in Groups A, B, C, D and E was 3.75, 1.15,1.12, 1.10 and 1.09 folds of the control group, respectively. The difference was only statistically significant between Group A and the control group ( P < 0. 01 ). Conclusion β-NGF released from the β-NGF controlled delivery system was bioactive. It could promote the growth of DRG axons.     

The effects of β-NGF controlled delivery system on growth of primary cultured chick embryo dorsal root ganglion axons

Objective To construct β-NGF controlled delivery system and evaluate the biological effects of β-NGF on the growth of chick embryo dorsal root ganglion (DRG) axons in vitro. Methods Delivery systems releasing β-NGF at 50/μL, 100/μg/L and 250 μg/L concentration were constructed. To determine the optimal dose response effects of NGF in the controlled delivery system, DRG were co-cultured with of β-NGF at above concentrations while using DRG basic culture as control. Axonal growth was observed. DRG were also cocultured with the components in the controlled delivery system to detect the effects on growth of DRG axons. The experiment was divided into 5 experimental groups and 1 control group: control group, DRG+ fibrin; Group A,DRG+ fibrin+ peptide + heparin + 100 μg/L β-NGF; Group B, DRG + fibrin + heparin + 100/μg/L β-NGF;Group C, DRG + fibrin + peptide + 100 μg/L β-NGF; Group D, DRG + fibrin + 100 μg/L β-NGF; Group E,DRG + fibrin + peptide + heparin. Results The growth of DRG axons in 50 μg/L, 100μg/L and 250/μg/Lconcentration of β-NGF controlled delivery system was 1.31 ( P > 0. 05), 3.78 ( P < 0. 01 ) and 3.05 ( P <0.01) folds of the control respectively. The growth of DRG axons in 100 μg/L group was significantly better comparing to that in 250 μg/L group. The growth of DRG axons in Groups A, B, C, D and E was 3.75, 1.15,1.12, 1.10 and 1.09 folds of the control group, respectively. The difference was only statistically significant between Group A and the control group ( P < 0. 01 ). Conclusion β-NGF released from the β-NGF controlled delivery system was bioactive. It could promote the growth of DRG axons.     

The effects of β-NGF controlled delivery system on growth of primary cultured chick embryo dorsal root ganglion axons

Objective To construct β-NGF controlled delivery system and evaluate the biological effects of β-NGF on the growth of chick embryo dorsal root ganglion (DRG) axons in vitro. Methods Delivery systems releasing β-NGF at 50/μL, 100/μg/L and 250 μg/L concentration were constructed. To determine the optimal dose response effects of NGF in the controlled delivery system, DRG were co-cultured with of β-NGF at above concentrations while using DRG basic culture as control. Axonal growth was observed. DRG were also cocultured with the components in the controlled delivery system to detect the effects on growth of DRG axons. The experiment was divided into 5 experimental groups and 1 control group: control group, DRG+ fibrin; Group A,DRG+ fibrin+ peptide + heparin + 100 μg/L β-NGF; Group B, DRG + fibrin + heparin + 100/μg/L β-NGF;Group C, DRG + fibrin + peptide + 100 μg/L β-NGF; Group D, DRG + fibrin + 100 μg/L β-NGF; Group E,DRG + fibrin + peptide + heparin. Results The growth of DRG axons in 50 μg/L, 100μg/L and 250/μg/Lconcentration of β-NGF controlled delivery system was 1.31 ( P > 0. 05), 3.78 ( P < 0. 01 ) and 3.05 ( P <0.01) folds of the control respectively. The growth of DRG axons in 100 μg/L group was significantly better comparing to that in 250 μg/L group. The growth of DRG axons in Groups A, B, C, D and E was 3.75, 1.15,1.12, 1.10 and 1.09 folds of the control group, respectively. The difference was only statistically significant between Group A and the control group ( P < 0. 01 ). Conclusion β-NGF released from the β-NGF controlled delivery system was bioactive. It could promote the growth of DRG axons.     

The effects of β-NGF controlled delivery system on growth of primary cultured chick embryo dorsal root ganglion axons

Objective To construct β-NGF controlled delivery system and evaluate the biological effects of β-NGF on the growth of chick embryo dorsal root ganglion (DRG) axons in vitro. Methods Delivery systems releasing β-NGF at 50/μL, 100/μg/L and 250 μg/L concentration were constructed. To determine the optimal dose response effects of NGF in the controlled delivery system, DRG were co-cultured with of β-NGF at above concentrations while using DRG basic culture as control. Axonal growth was observed. DRG were also cocultured with the components in the controlled delivery system to detect the effects on growth of DRG axons. The experiment was divided into 5 experimental groups and 1 control group: control group, DRG+ fibrin; Group A,DRG+ fibrin+ peptide + heparin + 100 μg/L β-NGF; Group B, DRG + fibrin + heparin + 100/μg/L β-NGF;Group C, DRG + fibrin + peptide + 100 μg/L β-NGF; Group D, DRG + fibrin + 100 μg/L β-NGF; Group E,DRG + fibrin + peptide + heparin. Results The growth of DRG axons in 50 μg/L, 100μg/L and 250/μg/Lconcentration of β-NGF controlled delivery system was 1.31 ( P > 0. 05), 3.78 ( P < 0. 01 ) and 3.05 ( P <0.01) folds of the control respectively. The growth of DRG axons in 100 μg/L group was significantly better comparing to that in 250 μg/L group. The growth of DRG axons in Groups A, B, C, D and E was 3.75, 1.15,1.12, 1.10 and 1.09 folds of the control group, respectively. The difference was only statistically significant between Group A and the control group ( P < 0. 01 ). Conclusion β-NGF released from the β-NGF controlled delivery system was bioactive. It could promote the growth of DRG axons.     

The effects of β-NGF controlled delivery system on growth of primary cultured chick embryo dorsal root ganglion axons

Objective To construct β-NGF controlled delivery system and evaluate the biological effects of β-NGF on the growth of chick embryo dorsal root ganglion (DRG) axons in vitro. Methods Delivery systems releasing β-NGF at 50/μL, 100/μg/L and 250 μg/L concentration were constructed. To determine the optimal dose response effects of NGF in the controlled delivery system, DRG were co-cultured with of β-NGF at above concentrations while using DRG basic culture as control. Axonal growth was observed. DRG were also cocultured with the components in the controlled delivery system to detect the effects on growth of DRG axons. The experiment was divided into 5 experimental groups and 1 control group: control group, DRG+ fibrin; Group A,DRG+ fibrin+ peptide + heparin + 100 μg/L β-NGF; Group B, DRG + fibrin + heparin + 100/μg/L β-NGF;Group C, DRG + fibrin + peptide + 100 μg/L β-NGF; Group D, DRG + fibrin + 100 μg/L β-NGF; Group E,DRG + fibrin + peptide + heparin. Results The growth of DRG axons in 50 μg/L, 100μg/L and 250/μg/Lconcentration of β-NGF controlled delivery system was 1.31 ( P > 0. 05), 3.78 ( P < 0. 01 ) and 3.05 ( P <0.01) folds of the control respectively. The growth of DRG axons in 100 μg/L group was significantly better comparing to that in 250 μg/L group. The growth of DRG axons in Groups A, B, C, D and E was 3.75, 1.15,1.12, 1.10 and 1.09 folds of the control group, respectively. The difference was only statistically significant between Group A and the control group ( P < 0. 01 ). Conclusion β-NGF released from the β-NGF controlled delivery system was bioactive. It could promote the growth of DRG axons.     

The effects of β-NGF controlled delivery system on growth of primary cultured chick embryo dorsal root ganglion axons

Objective To construct β-NGF controlled delivery system and evaluate the biological effects of β-NGF on the growth of chick embryo dorsal root ganglion (DRG) axons in vitro. Methods Delivery systems releasing β-NGF at 50/μL, 100/μg/L and 250 μg/L concentration were constructed. To determine the optimal dose response effects of NGF in the controlled delivery system, DRG were co-cultured with of β-NGF at above concentrations while using DRG basic culture as control. Axonal growth was observed. DRG were also cocultured with the components in the controlled delivery system to detect the effects on growth of DRG axons. The experiment was divided into 5 experimental groups and 1 control group: control group, DRG+ fibrin; Group A,DRG+ fibrin+ peptide + heparin + 100 μg/L β-NGF; Group B, DRG + fibrin + heparin + 100/μg/L β-NGF;Group C, DRG + fibrin + peptide + 100 μg/L β-NGF; Group D, DRG + fibrin + 100 μg/L β-NGF; Group E,DRG + fibrin + peptide + heparin. Results The growth of DRG axons in 50 μg/L, 100μg/L and 250/μg/Lconcentration of β-NGF controlled delivery system was 1.31 ( P > 0. 05), 3.78 ( P < 0. 01 ) and 3.05 ( P <0.01) folds of the control respectively. The growth of DRG axons in 100 μg/L group was significantly better comparing to that in 250 μg/L group. The growth of DRG axons in Groups A, B, C, D and E was 3.75, 1.15,1.12, 1.10 and 1.09 folds of the control group, respectively. The difference was only statistically significant between Group A and the control group ( P < 0. 01 ). Conclusion β-NGF released from the β-NGF controlled delivery system was bioactive. It could promote the growth of DRG axons.     

The effects of β-NGF controlled delivery system on growth of primary cultured chick embryo dorsal root ganglion axons

Objective To construct β-NGF controlled delivery system and evaluate the biological effects of β-NGF on the growth of chick embryo dorsal root ganglion (DRG) axons in vitro. Methods Delivery systems releasing β-NGF at 50/μL, 100/μg/L and 250 μg/L concentration were constructed. To determine the optimal dose response effects of NGF in the controlled delivery system, DRG were co-cultured with of β-NGF at above concentrations while using DRG basic culture as control. Axonal growth was observed. DRG were also cocultured with the components in the controlled delivery system to detect the effects on growth of DRG axons. The experiment was divided into 5 experimental groups and 1 control group: control group, DRG+ fibrin; Group A,DRG+ fibrin+ peptide + heparin + 100 μg/L β-NGF; Group B, DRG + fibrin + heparin + 100/μg/L β-NGF;Group C, DRG + fibrin + peptide + 100 μg/L β-NGF; Group D, DRG + fibrin + 100 μg/L β-NGF; Group E,DRG + fibrin + peptide + heparin. Results The growth of DRG axons in 50 μg/L, 100μg/L and 250/μg/Lconcentration of β-NGF controlled delivery system was 1.31 ( P > 0. 05), 3.78 ( P < 0. 01 ) and 3.05 ( P <0.01) folds of the control respectively. The growth of DRG axons in 100 μg/L group was significantly better comparing to that in 250 μg/L group. The growth of DRG axons in Groups A, B, C, D and E was 3.75, 1.15,1.12, 1.10 and 1.09 folds of the control group, respectively. The difference was only statistically significant between Group A and the control group ( P < 0. 01 ). Conclusion β-NGF released from the β-NGF controlled delivery system was bioactive. It could promote the growth of DRG axons.     

神经生长因子对胆管癌细胞67-kDa层黏连蛋白受体表达的影响

Objective To investigate the effects of nerve growth factor (NGF) on the expression of 67-kDa laminin receptor (67LR) in human bile duct carcinoma QBC939 cells, and study the possible mechanism of perineural invasion and metastasis of bile duct carcinoma. Methods ( 1 ) The expression of a high-affinity receptor for NGF, TrkA, was detected by immunofluorescence staining. ( 2 ) QBC393 cells were pretreated by β-NGF at different concentrations ( 1, 10, 100,200 μg/L), and then the mRNA and protein expressions of 67LR were examined by Real-Time PCR and Western blot assay. QBC939 cells were divided into control group and β-NGF (1, 10, 100,200 μg/L) groups. (3) The ideal concentration of β-NGF was selected according to the results of previous tests, and then the mRNA and protein expressions of 67LR were re-examined by adding specific TrkA inhibitor K252a at different concentrations ( 100,200,300 nmol/L). QBC939 cells were divided into control group, β-NGF 100 μg/L group and K252a ( 100,200,300 nmol/L) groups. All data were analyzed by one-way analysis of variance or LSD-test. Results (1) A strong expression of TrkA was detected in the membrane of QBC939 cells. (2) The mRNA and protein expressions of 67LR in QBC939 cells were 0.35 ± 0.06 and 0. 32 ± 0.05 in the control group, 0.38 ±0.14 and 0.50 ±0.09 in the β-NGF 1 μg/L group, 0.62 ±0.14 and 0. 69 ±0. 13 in β-NGF 10 μg/L group, 0.90 ± 0.08 and 0.93 ± 0.07 in the β-NGF 100 μg/L group, and 0. 70 ± 0. 10 and 0. 76 ±0.07 in the β-NGF 200 μg/L group, there were significant differences among the five groups (F = 22. 4, 14. 6,P ＜0.05). The mRNA and protein expressions of 67LR in the β-NGF 100 μg/L group were significantly higher than those in the control group ( t = 19. 0, 21.0, P ＜ 0. 05 ). (3) The mRNA and protein expressions of 67LR in the QBC939 cells were 0.35 ±0.10 and 0.41 ±0.10 in the control group, 0. 88 ±0. 14 and 0.84 ±0.10 in the β-NGF 100 μg/L group, 0.80±0.08 and 0.76 ±0.04 in the K252a 100 nmol/L group, 0.67 ±0.12 and 0.61 ± 0.09 in the K252a 200 nmol/L group, and 0. 43 ± 0.07 and 0. 50 ± 0. 12 in the K252a 300 nmol/L group, there were significant differences among the five groups ( F = 14. 1, 8. 9, P ＜ 0.05 ). There were no significant differences in the mRNA and protein expressions of 67LR between the K252a 300 nmol/L group and the control group (t =1.02, 0. 85, P＞0.05). Conclusion In bile duct carcinoma cells, NGF enhances the expression of 67LR by combining with TrkA, which might be the mechanism of NGF mediating perineural invasion of bile duct carcinoma.     

神经生长因子对胆管癌细胞67-kDa层黏连蛋白受体表达的影响

目的 研究外源性神经生长因子(NGF)对QBC939细胞67-kDa层黏连蛋白受体(67LR)表达的影响,探讨胆管癌神经浸润的机制.方法 (1)细胞免疫荧光染色法检测QBC939细胞表面NGF高亲和力酪氨酸激酶受体A(TrkA)表达情况.(2)用不同浓度的重组人神经生长因子(β-NGF)对QBC939细胞进行处理,采用Real-Time PCR和Western blot检测QBC393细胞67LR mRNA和蛋白表达情况.实验分为对照组、β-NGF(1、10、100、200μg/L)组.(3)根据上述实验结果选定最佳β-NGF浓度,再加入不同浓度的TrkA阻断剂K252a,再次检测QBC939细胞67LR mRNA和蛋白表达情况.实验分为对照组、100μg/Lβ-NGF组、K252a(100、200、300 nmol/L)阻断组.数据采用单因素方差分析,两两比较采用LSD法.结果 (1)QBC939细胞膜上TrkA表达明显.(2)对照组和β-NGF(1、10、100、200 μg/L)组QBC939细胞67LRmRNA和蛋白表达水平分别为0.35±0.06、0.38±0.14、0.62±0.14、0.90±0.08、0.70±0.10和0.32±0.05、0.50±0.09、0.69±0.13、0.93±0.07、0.76±0.07,两者比较,差异有统计学意义(F=22.4,14.6,P＜0.05),其中100ng/ml β-NGF作用最明显(t=19.0,21.0,P＜0.05).(3)对照组、100 μg/L β-NGF组、K252a(100、200、300 nmol/L)阻断组QBC939细胞67LR mRNA和蛋白表达水平分别为0.35±0.10、0.88±0.14、0.80±0.08、0.67±0.12、0.43±0.07和0.41±0.10、0.84±0.10、0.76±0.04、0.61±0.09、0.50±0.12,两者比较,差异有统计学意义(F=14.1,8.9,P＜0.05),其中经300 nmol/L K252a处理后,QBC939细胞67LR mRNA和蛋白表达水平与对照组比较,差异无统计学意义(t=1.02,0.85,P＞0.05).结论 外源性NGF通过与其高亲和力受体TrkA结合,可增强QBC939细胞67LR表达,这可能是促进胆管癌细胞延外周神经纤维间隙浸润的机制之一.

Abstract：

Objective To investigate the effects of nerve growth factor (NGF) on the expression of 67-kDa laminin receptor (67LR) in human bile duct carcinoma QBC939 cells, and study the possible mechanism of perineural invasion and metastasis of bile duct carcinoma. Methods ( 1 ) The expression of a high-affinity receptor for NGF, TrkA, was detected by immunofluorescence staining. ( 2 ) QBC393 cells were pretreated by β-NGF at different concentrations ( 1, 10, 100,200 μg/L), and then the mRNA and protein expressions of 67LR were examined by Real-Time PCR and Western blot assay. QBC939 cells were divided into control group and β-NGF (1, 10, 100,200 μg/L) groups. (3) The ideal concentration of β-NGF was selected according to the results of previous tests, and then the mRNA and protein expressions of 67LR were re-examined by adding specific TrkA inhibitor K252a at different concentrations ( 100,200,300 nmol/L). QBC939 cells were divided into control group, β-NGF 100 μg/L group and K252a ( 100,200,300 nmol/L) groups. All data were analyzed by one-way analysis of variance or LSD-test. Results (1) A strong expression of TrkA was detected in the membrane of QBC939 cells. (2) The mRNA and protein expressions of 67LR in QBC939 cells were 0.35 ± 0.06 and 0. 32 ± 0.05 in the control group, 0.38 ±0.14 and 0.50 ±0.09 in the β-NGF 1 μg/L group, 0.62 ±0.14 and 0. 69 ±0. 13 in β-NGF 10 μg/L group, 0.90 ± 0.08 and 0.93 ± 0.07 in the β-NGF 100 μg/L group, and 0. 70 ± 0. 10 and 0. 76 ±0.07 in the β-NGF 200 μg/L group, there were significant differences among the five groups (F = 22. 4, 14. 6,P ＜0.05). The mRNA and protein expressions of 67LR in the β-NGF 100 μg/L group were significantly higher than those in the control group ( t = 19. 0, 21.0, P ＜ 0. 05 ). (3) The mRNA and protein expressions of 67LR in the QBC939 cells were 0.35 ±0.10 and 0.41 ±0.10 in the control group, 0. 88 ±0. 14 and 0.84 ±0.10 in the β-NGF 100 μg/L group, 0.80±0.08 and 0.76 ±0.04 in the K252a 100 nmol/L group, 0.67 ±0.12 and 0.61 ± 0.09 in the K252a 200 nmol/L group, and 0. 43 ± 0.07 and 0. 50 ± 0. 12 in the K252a 300 nmol/L group, there were significant differences among the five groups ( F = 14. 1, 8. 9, P ＜ 0.05 ). There were no significant differences in the mRNA and protein expressions of 67LR between the K252a 300 nmol/L group and the control group (t =1.02, 0. 85, P＞0.05). Conclusion In bile duct carcinoma cells, NGF enhances the expression of 67LR by combining with TrkA, which might be the mechanism of NGF mediating perineural invasion of bile duct carcinoma.     

The expression of β-adrenoceptors in human pancreatic cancer cell lines and their roles in cell invasiveness

Objective To investigate the effect and mechanism of β-adrenoceptors on norepi-nephrine-induced invasiveness of pancreatic cancer cell lines. Methods The expression of β-adrenocep-tors mRNA in human pancreatic cancer cell lines MiaPaCa-2 and BxPC3 was detected by using RT-PCR. The cells were randomly divided into control group.10 mol/L NE intervention group, 1 mol/L propranolol intervention group and NE + propranolol intervention group. After 48 h , transwell invasiveness test was used to examine the changes in invasive ability of MiaPaCa-2. The expression of MMP-2, MMP-9 and VEGF mRNA was measured by semi-quantitative RT-PCR. The levels of MMP-2 , MMP-9 and VECF proteins were assayed by immunocytochemistry. Results Both MiaPaCa-2 and BxPC3 expressed β1-and β2-adrenocep-tors. The absorbance ( A) values of invasive cells in NE, NE + propranolol, propranolol and control groups were 0.78±0.02 ,0.32±0.03 ,0.26±0.01 and 0.28±0.02 , respectively, and those in NE intervention group were significantly higher than in control and NE + propranolol groups ( P <0.05) . There was no sig-nificant difference in the number of invasive cells between propranolol and control groups ( P > 0. 05) . In NE group , the expression index of MMP-2 , MMP-9 and VEGF mRNA was 0. 87±0.02 , 1.04±0.02 and 0. 92±0. 01 , and the gray value of the protein expression was 131.20±2.34,105.32±7.21 and 115.60 ±5. 03 , respectively, which were higher than those in control and NE + propranolol groups ( P<0.05). There was no significant difference in the expression levels of MMP-2 , MMP-9 and VEGF mRNA and pro-tein between propranolol and control groups ( P>0.05 ) . Conclusion β-adrenoceptors play an important role in the process of norepinephrine-induced invasiveness of pancreatic cancer cells. NE can promote the invasiveness of MiaPaCa-2 through up-regulating the expression of MMP-2 , MMP-9 and VEGF via β-adre-noceptors.     

The expression of β-adrenoceptors in human pancreatic cancer cell lines and their roles in cell invasiveness

Objective To investigate the effect and mechanism of β-adrenoceptors on norepi-nephrine-induced invasiveness of pancreatic cancer cell lines. Methods The expression of β-adrenocep-tors mRNA in human pancreatic cancer cell lines MiaPaCa-2 and BxPC3 was detected by using RT-PCR. The cells were randomly divided into control group.10 mol/L NE intervention group, 1 mol/L propranolol intervention group and NE + propranolol intervention group. After 48 h , transwell invasiveness test was used to examine the changes in invasive ability of MiaPaCa-2. The expression of MMP-2, MMP-9 and VEGF mRNA was measured by semi-quantitative RT-PCR. The levels of MMP-2 , MMP-9 and VECF proteins were assayed by immunocytochemistry. Results Both MiaPaCa-2 and BxPC3 expressed β1-and β2-adrenocep-tors. The absorbance ( A) values of invasive cells in NE, NE + propranolol, propranolol and control groups were 0.78±0.02 ,0.32±0.03 ,0.26±0.01 and 0.28±0.02 , respectively, and those in NE intervention group were significantly higher than in control and NE + propranolol groups ( P <0.05) . There was no sig-nificant difference in the number of invasive cells between propranolol and control groups ( P > 0. 05) . In NE group , the expression index of MMP-2 , MMP-9 and VEGF mRNA was 0. 87±0.02 , 1.04±0.02 and 0. 92±0. 01 , and the gray value of the protein expression was 131.20±2.34,105.32±7.21 and 115.60 ±5. 03 , respectively, which were higher than those in control and NE + propranolol groups ( P<0.05). There was no significant difference in the expression levels of MMP-2 , MMP-9 and VEGF mRNA and pro-tein between propranolol and control groups ( P>0.05 ) . Conclusion β-adrenoceptors play an important role in the process of norepinephrine-induced invasiveness of pancreatic cancer cells. NE can promote the invasiveness of MiaPaCa-2 through up-regulating the expression of MMP-2 , MMP-9 and VEGF via β-adre-noceptors.     

The expression of β-adrenoceptors in human pancreatic cancer cell lines and their roles in cell invasiveness

Objective To investigate the effect and mechanism of β-adrenoceptors on norepi-nephrine-induced invasiveness of pancreatic cancer cell lines. Methods The expression of β-adrenocep-tors mRNA in human pancreatic cancer cell lines MiaPaCa-2 and BxPC3 was detected by using RT-PCR. The cells were randomly divided into control group.10 mol/L NE intervention group, 1 mol/L propranolol intervention group and NE + propranolol intervention group. After 48 h , transwell invasiveness test was used to examine the changes in invasive ability of MiaPaCa-2. The expression of MMP-2, MMP-9 and VEGF mRNA was measured by semi-quantitative RT-PCR. The levels of MMP-2 , MMP-9 and VECF proteins were assayed by immunocytochemistry. Results Both MiaPaCa-2 and BxPC3 expressed β1-and β2-adrenocep-tors. The absorbance ( A) values of invasive cells in NE, NE + propranolol, propranolol and control groups were 0.78±0.02 ,0.32±0.03 ,0.26±0.01 and 0.28±0.02 , respectively, and those in NE intervention group were significantly higher than in control and NE + propranolol groups ( P <0.05) . There was no sig-nificant difference in the number of invasive cells between propranolol and control groups ( P > 0. 05) . In NE group , the expression index of MMP-2 , MMP-9 and VEGF mRNA was 0. 87±0.02 , 1.04±0.02 and 0. 92±0. 01 , and the gray value of the protein expression was 131.20±2.34,105.32±7.21 and 115.60 ±5. 03 , respectively, which were higher than those in control and NE + propranolol groups ( P<0.05). There was no significant difference in the expression levels of MMP-2 , MMP-9 and VEGF mRNA and pro-tein between propranolol and control groups ( P>0.05 ) . Conclusion β-adrenoceptors play an important role in the process of norepinephrine-induced invasiveness of pancreatic cancer cells. NE can promote the invasiveness of MiaPaCa-2 through up-regulating the expression of MMP-2 , MMP-9 and VEGF via β-adre-noceptors.     

The expression of β-adrenoceptors in human pancreatic cancer cell lines and their roles in cell invasiveness

Objective To investigate the effect and mechanism of β-adrenoceptors on norepi-nephrine-induced invasiveness of pancreatic cancer cell lines. Methods The expression of β-adrenocep-tors mRNA in human pancreatic cancer cell lines MiaPaCa-2 and BxPC3 was detected by using RT-PCR. The cells were randomly divided into control group.10 mol/L NE intervention group, 1 mol/L propranolol intervention group and NE + propranolol intervention group. After 48 h , transwell invasiveness test was used to examine the changes in invasive ability of MiaPaCa-2. The expression of MMP-2, MMP-9 and VEGF mRNA was measured by semi-quantitative RT-PCR. The levels of MMP-2 , MMP-9 and VECF proteins were assayed by immunocytochemistry. Results Both MiaPaCa-2 and BxPC3 expressed β1-and β2-adrenocep-tors. The absorbance ( A) values of invasive cells in NE, NE + propranolol, propranolol and control groups were 0.78±0.02 ,0.32±0.03 ,0.26±0.01 and 0.28±0.02 , respectively, and those in NE intervention group were significantly higher than in control and NE + propranolol groups ( P <0.05) . There was no sig-nificant difference in the number of invasive cells between propranolol and control groups ( P > 0. 05) . In NE group , the expression index of MMP-2 , MMP-9 and VEGF mRNA was 0. 87±0.02 , 1.04±0.02 and 0. 92±0. 01 , and the gray value of the protein expression was 131.20±2.34,105.32±7.21 and 115.60 ±5. 03 , respectively, which were higher than those in control and NE + propranolol groups ( P<0.05). There was no significant difference in the expression levels of MMP-2 , MMP-9 and VEGF mRNA and pro-tein between propranolol and control groups ( P>0.05 ) . Conclusion β-adrenoceptors play an important role in the process of norepinephrine-induced invasiveness of pancreatic cancer cells. NE can promote the invasiveness of MiaPaCa-2 through up-regulating the expression of MMP-2 , MMP-9 and VEGF via β-adre-noceptors.     

The expression of β-adrenoceptors in human pancreatic cancer cell lines and their roles in cell invasiveness

Objective To investigate the effect and mechanism of β-adrenoceptors on norepi-nephrine-induced invasiveness of pancreatic cancer cell lines. Methods The expression of β-adrenocep-tors mRNA in human pancreatic cancer cell lines MiaPaCa-2 and BxPC3 was detected by using RT-PCR. The cells were randomly divided into control group.10 mol/L NE intervention group, 1 mol/L propranolol intervention group and NE + propranolol intervention group. After 48 h , transwell invasiveness test was used to examine the changes in invasive ability of MiaPaCa-2. The expression of MMP-2, MMP-9 and VEGF mRNA was measured by semi-quantitative RT-PCR. The levels of MMP-2 , MMP-9 and VECF proteins were assayed by immunocytochemistry. Results Both MiaPaCa-2 and BxPC3 expressed β1-and β2-adrenocep-tors. The absorbance ( A) values of invasive cells in NE, NE + propranolol, propranolol and control groups were 0.78±0.02 ,0.32±0.03 ,0.26±0.01 and 0.28±0.02 , respectively, and those in NE intervention group were significantly higher than in control and NE + propranolol groups ( P <0.05) . There was no sig-nificant difference in the number of invasive cells between propranolol and control groups ( P > 0. 05) . In NE group , the expression index of MMP-2 , MMP-9 and VEGF mRNA was 0. 87±0.02 , 1.04±0.02 and 0. 92±0. 01 , and the gray value of the protein expression was 131.20±2.34,105.32±7.21 and 115.60 ±5. 03 , respectively, which were higher than those in control and NE + propranolol groups ( P<0.05). There was no significant difference in the expression levels of MMP-2 , MMP-9 and VEGF mRNA and pro-tein between propranolol and control groups ( P>0.05 ) . Conclusion β-adrenoceptors play an important role in the process of norepinephrine-induced invasiveness of pancreatic cancer cells. NE can promote the invasiveness of MiaPaCa-2 through up-regulating the expression of MMP-2 , MMP-9 and VEGF via β-adre-noceptors.     

The expression of β-adrenoceptors in human pancreatic cancer cell lines and their roles in cell invasiveness

Objective To investigate the effect and mechanism of β-adrenoceptors on norepi-nephrine-induced invasiveness of pancreatic cancer cell lines. Methods The expression of β-adrenocep-tors mRNA in human pancreatic cancer cell lines MiaPaCa-2 and BxPC3 was detected by using RT-PCR. The cells were randomly divided into control group.10 mol/L NE intervention group, 1 mol/L propranolol intervention group and NE + propranolol intervention group. After 48 h , transwell invasiveness test was used to examine the changes in invasive ability of MiaPaCa-2. The expression of MMP-2, MMP-9 and VEGF mRNA was measured by semi-quantitative RT-PCR. The levels of MMP-2 , MMP-9 and VECF proteins were assayed by immunocytochemistry. Results Both MiaPaCa-2 and BxPC3 expressed β1-and β2-adrenocep-tors. The absorbance ( A) values of invasive cells in NE, NE + propranolol, propranolol and control groups were 0.78±0.02 ,0.32±0.03 ,0.26±0.01 and 0.28±0.02 , respectively, and those in NE intervention group were significantly higher than in control and NE + propranolol groups ( P <0.05) . There was no sig-nificant difference in the number of invasive cells between propranolol and control groups ( P > 0. 05) . In NE group , the expression index of MMP-2 , MMP-9 and VEGF mRNA was 0. 87±0.02 , 1.04±0.02 and 0. 92±0. 01 , and the gray value of the protein expression was 131.20±2.34,105.32±7.21 and 115.60 ±5. 03 , respectively, which were higher than those in control and NE + propranolol groups ( P<0.05). There was no significant difference in the expression levels of MMP-2 , MMP-9 and VEGF mRNA and pro-tein between propranolol and control groups ( P>0.05 ) . Conclusion β-adrenoceptors play an important role in the process of norepinephrine-induced invasiveness of pancreatic cancer cells. NE can promote the invasiveness of MiaPaCa-2 through up-regulating the expression of MMP-2 , MMP-9 and VEGF via β-adre-noceptors.     

The expression of β-adrenoceptors in human pancreatic cancer cell lines and their roles in cell invasiveness

Objective To investigate the effect and mechanism of β-adrenoceptors on norepi-nephrine-induced invasiveness of pancreatic cancer cell lines. Methods The expression of β-adrenocep-tors mRNA in human pancreatic cancer cell lines MiaPaCa-2 and BxPC3 was detected by using RT-PCR. The cells were randomly divided into control group.10 mol/L NE intervention group, 1 mol/L propranolol intervention group and NE + propranolol intervention group. After 48 h , transwell invasiveness test was used to examine the changes in invasive ability of MiaPaCa-2. The expression of MMP-2, MMP-9 and VEGF mRNA was measured by semi-quantitative RT-PCR. The levels of MMP-2 , MMP-9 and VECF proteins were assayed by immunocytochemistry. Results Both MiaPaCa-2 and BxPC3 expressed β1-and β2-adrenocep-tors. The absorbance ( A) values of invasive cells in NE, NE + propranolol, propranolol and control groups were 0.78±0.02 ,0.32±0.03 ,0.26±0.01 and 0.28±0.02 , respectively, and those in NE intervention group were significantly higher than in control and NE + propranolol groups ( P <0.05) . There was no sig-nificant difference in the number of invasive cells between propranolol and control groups ( P > 0. 05) . In NE group , the expression index of MMP-2 , MMP-9 and VEGF mRNA was 0. 87±0.02 , 1.04±0.02 and 0. 92±0. 01 , and the gray value of the protein expression was 131.20±2.34,105.32±7.21 and 115.60 ±5. 03 , respectively, which were higher than those in control and NE + propranolol groups ( P<0.05). There was no significant difference in the expression levels of MMP-2 , MMP-9 and VEGF mRNA and pro-tein between propranolol and control groups ( P>0.05 ) . Conclusion β-adrenoceptors play an important role in the process of norepinephrine-induced invasiveness of pancreatic cancer cells. NE can promote the invasiveness of MiaPaCa-2 through up-regulating the expression of MMP-2 , MMP-9 and VEGF via β-adre-noceptors.     

The expression of β-adrenoceptors in human pancreatic cancer cell lines and their roles in cell invasiveness

Objective To investigate the effect and mechanism of β-adrenoceptors on norepi-nephrine-induced invasiveness of pancreatic cancer cell lines. Methods The expression of β-adrenocep-tors mRNA in human pancreatic cancer cell lines MiaPaCa-2 and BxPC3 was detected by using RT-PCR. The cells were randomly divided into control group.10 mol/L NE intervention group, 1 mol/L propranolol intervention group and NE + propranolol intervention group. After 48 h , transwell invasiveness test was used to examine the changes in invasive ability of MiaPaCa-2. The expression of MMP-2, MMP-9 and VEGF mRNA was measured by semi-quantitative RT-PCR. The levels of MMP-2 , MMP-9 and VECF proteins were assayed by immunocytochemistry. Results Both MiaPaCa-2 and BxPC3 expressed β1-and β2-adrenocep-tors. The absorbance ( A) values of invasive cells in NE, NE + propranolol, propranolol and control groups were 0.78±0.02 ,0.32±0.03 ,0.26±0.01 and 0.28±0.02 , respectively, and those in NE intervention group were significantly higher than in control and NE + propranolol groups ( P <0.05) . There was no sig-nificant difference in the number of invasive cells between propranolol and control groups ( P > 0. 05) . In NE group , the expression index of MMP-2 , MMP-9 and VEGF mRNA was 0. 87±0.02 , 1.04±0.02 and 0. 92±0. 01 , and the gray value of the protein expression was 131.20±2.34,105.32±7.21 and 115.60 ±5. 03 , respectively, which were higher than those in control and NE + propranolol groups ( P<0.05). There was no significant difference in the expression levels of MMP-2 , MMP-9 and VEGF mRNA and pro-tein between propranolol and control groups ( P>0.05 ) . Conclusion β-adrenoceptors play an important role in the process of norepinephrine-induced invasiveness of pancreatic cancer cells. NE can promote the invasiveness of MiaPaCa-2 through up-regulating the expression of MMP-2 , MMP-9 and VEGF via β-adre-noceptors.     

The expression of β-adrenoceptors in human pancreatic cancer cell lines and their roles in cell invasiveness

Objective To investigate the effect and mechanism of β-adrenoceptors on norepi-nephrine-induced invasiveness of pancreatic cancer cell lines. Methods The expression of β-adrenocep-tors mRNA in human pancreatic cancer cell lines MiaPaCa-2 and BxPC3 was detected by using RT-PCR. The cells were randomly divided into control group.10 mol/L NE intervention group, 1 mol/L propranolol intervention group and NE + propranolol intervention group. After 48 h , transwell invasiveness test was used to examine the changes in invasive ability of MiaPaCa-2. The expression of MMP-2, MMP-9 and VEGF mRNA was measured by semi-quantitative RT-PCR. The levels of MMP-2 , MMP-9 and VECF proteins were assayed by immunocytochemistry. Results Both MiaPaCa-2 and BxPC3 expressed β1-and β2-adrenocep-tors. The absorbance ( A) values of invasive cells in NE, NE + propranolol, propranolol and control groups were 0.78±0.02 ,0.32±0.03 ,0.26±0.01 and 0.28±0.02 , respectively, and those in NE intervention group were significantly higher than in control and NE + propranolol groups ( P <0.05) . There was no sig-nificant difference in the number of invasive cells between propranolol and control groups ( P > 0. 05) . In NE group , the expression index of MMP-2 , MMP-9 and VEGF mRNA was 0. 87±0.02 , 1.04±0.02 and 0. 92±0. 01 , and the gray value of the protein expression was 131.20±2.34,105.32±7.21 and 115.60 ±5. 03 , respectively, which were higher than those in control and NE + propranolol groups ( P<0.05). There was no significant difference in the expression levels of MMP-2 , MMP-9 and VEGF mRNA and pro-tein between propranolol and control groups ( P>0.05 ) . Conclusion β-adrenoceptors play an important role in the process of norepinephrine-induced invasiveness of pancreatic cancer cells. NE can promote the invasiveness of MiaPaCa-2 through up-regulating the expression of MMP-2 , MMP-9 and VEGF via β-adre-noceptors.