Nerve Growth Factor on the Proliferation of Human Pancreatic Carcinoma Cells
0
Citation
0
Reference
20
Related Paper
Abstract:
BACKGROUND AND AIM:To investigate the effect of nerve growth factor(β-NGF) on the proliferative ability and cell cycle of human pancreatic carcinoma cells MIA PaCa-2.MATERIALS AND METHODS:Colony forming efficiency of MIA PaCa-2 cells treated with β-NGF was higher than normal group(P0.05),especially with the 100 ng/ml β-NGF group.Absorbence gradually increased,along with β-NGF concentration enhancement and culture time prolongation.Proliferation rate reached 48.68% when cultured 72 hours in 100 ng/ml β-NGF group.However,absorbence gradually decreased in K252a group at 24,48,72 h and 96 h.Inhibition rate was reduced to 18.05% in 100 ng/ml K252a group.Effect of different concentrations of β-NGF and K252a on MIA PaCa-2 cells proliferation was evaluated by MTT and Flow Cytometry.RESULTS:β-NGF promoted MIA PaCa-2 proliferation and growth which were markedly inhibited by K252a(P0.05).CONCLUSION:β-NGF could enhance the proliferation of MIA PaCa-2 pancreatic carcinoma cells.Keywords:
Pancreatic carcinoma
MTT assay
Cite
Objective To evaluate the effects of somatostatin analogue (octreotide) on the expression ofvascular endothelial growth factor (VEGF), cell proliferation, and apoptosis in hepatoma carcinoma cells. Methods 40 male mice which had received subcutaneous injection of murine hepatoma carcinoma cells were divided into 2 groups(each n=20). Group A received peritoneal injection of 100 μg/(kg·d) of octreotide for 20 days and Group B receivedperitoneal injection of 100 μg / (kg·d) of normal saline for 14 days 24 h later. All mice were sacrificed 20 days later,and the tumor size and the expression of VEGF were tested. Hepatoma carcinoma cell line SMMC-7721 was cultured invitro and received 10 μg/mL of octreotide arrangement, after being digested by RNase and stained by DNA-Stain, thecontent of DNA was measured by flow cytometry and the cell cycle were analyzed to calculate the apoptosis rate.Results The size of subcutaneous tumor was significantly smaller and the expression of VEGF was significantly lower inGroup A than those in Group B at day 7 and day 14 after implantation respectively (P0.05 or P0.01).Microvessel density in Group A was lower than that in Group B, but without significant difference (P=0.075). SMMC-7721 cells stayed in G0/G1 stage after being treated with octreotide, the cells in S and G2/M stage were decreased,and the apoptosis rate was increased. Conclusion Octreotide can inhibit the expression of VEGF, decrease theproliferation, and induce the apoptosis of hepatocellular carcinoma cells.
Subcutaneous injection
Cite
Citations (0)
Objective To investigate the effect of Iodine 125 seeds short time low dose rate irradiation on perineural invasion (PNI) in pancreatic cancer Capan-2 cells,and explore its molecular mechanism.Methods The co-culture model was established by co-culturing the dorsal root ganglion (DRG) of SD rat and Capzn-2 cells line,while Capan-2 culture model and DRG culture model was also established.Iodine 125 seeds short time low dose rate irradiation tablet was used for the 3 models,and the model without irradiation was used as control.Cancer cell and DRG growth was observed under inverted microscopy,surface of neurite and cell colony growth was determined by image analysis software.The concentration of nerve growth factor (NGF),transforming growth factor-α (TGF-α) in cell culture supernatant and matrigel solution was tested by ELISA,and the expression of neurotrophin-3 (NT-3) mRNA was detected by semi-quantitative reverse transcriptase polymerase chain reaction (RT-PCR).Results In the co-culture model,neurite of DRG showed a direction to cancer cells and had a concentrated growth towards cancer cells.And Capan-2 cells formed more colonies towards neurite.However,in irradiation groups,the symbiotic phenomenon was inhibited to some degree.Increased surface of neurite in co-culture model at 5th day was 290.15 ± 12.08,which was significantly higher than that in DRG group (124.83 ± 6.96,P ＜ 0.01),but the surface of neurite was decreased to 201.53 ± 12.20 after irradiation (P ＜0.01).Increased surface of Capan-2 cell was 300.47 ± 12.99,which was significantly higher than that in Capan-2 group (199.30 ± 8.60,P ＜ 0.01),but the surface of Capan-2 was decreased to 202.35 ± 7.97 after irradiation (P ＜ 0.01).NT-3 mRNA was seldom or not expressed in supernatant of co-culture model,but it was strongly expressed (0.68 ± 0.04) after irradiation (P ＜ 0.05).The concentration of NGF and TGF-α in supernatant of co-culture model were (27.56 ± 13.73),(40.86 ± 20.73) ng/ml,after irradiation they were increased to (94.98 ± 33.80),(157.54 ± 83.76) ng,/ml,and the difference between the two groups was statistically significant (P＜0.05 or ＜0.01).The concentration of NGF and TGF-α in matrigel lysate of co-culture model were (60.42 ± 33.03),(64.39 ± 21.52)ng/ml,after irradiation they were increased to (132.52 ±53.01),(138.38 ±83.58)ng/ml,and the difference of NGF concentration between the two groups was statistically significant (P ＜ 0.05).Conclusions Iodine-125 seeds short-time low-dose rate irradiation could inhibit interactions between nerve and Capan-2 cells,and the mechanism may be related to up-regulation of cancer cells perineural invasion promoter NGF,TGF-α and NT-3.
Key words:
Pancreatic neoplasms ; Iodine isotopes ; Low-level irradiation ; Neoplasm invasiveness
Neurite
Dorsal root ganglion
Matrigel
Cite
Citations (0)
Objective To investigate the effects of Plumbapin on the proliferation and apoptosis of tongue cancer cell line Tca.Methods The tongue cancer cells were put into 96-well plates(5×104 cells /well) and cultivated at 37℃,5% CO2 for 48 h.When cells grew to 80% confluence of the plate,the Plumbapin was added into each group respectively(with 0.025,0.1,0.2,0.3 and 0.4 mg/ml).After the treated cell cultured for 48,72,96 and 120 h,cell proliferation was evaluated by ELISA Reader at 490 nm with MTT assay.At mean time,apoptotic cell was detected by immunofluorescence technique with Hoechst33342 staining.Results Inhibition rate of tongue cancer cell was obviously enhanced with the increase of Plumbapin concentration.When the cells were treated with Plumbapin(0.4 mg/ml) for 48 h,statistical significance was shown compared with the other treated groups(P0.05).When the cells were cultured with Plumbapin(0.3 mg/ml)for 96h,cell apoptosis was significantly decreased compared with the groups of less than 0.3 mg/ml(P0.05).Hoechst33342 staining revealed that with the concentration increased,cell apoptosis rate was raised after 72 h.When the concentration of Plumbapin over 0.4 g/ml,complete nucleus was seldom seen under Fluorescence microscope instead of a large number of incomplete cell nucleus,nucleus debris and apoptotic body.Conclusion The Plumbapin can effectively inhibit tumor cell growth and induce tumor cell apoptosis.
Cell counting
Immunofluorescence
Cite
Citations (1)
Platelet-derived growth factor
MTT assay
Cite
Citations (0)
Objective To investigate the effect on proliferation and apoptosis of vascular endothelial cells exposed to high glucose. Methods The cultured human umbilical vein endothelial cells (HUVEC) were treated with high glucose at concentrations of 10,20,30,40,50 mmol/L for 2,4,6,8,10,12,14 days,cpm value was studied by using tritium-labelled thymidine deoxyribose( 3H-TdR)incorporation assay.Flow cytometry was used to detect apoptosis index,proliferation index and cell cycle. Results Treated with high glucose,the proliferation index and cpm were significantly reduced in a dose and time dependent manner than the control groups(P0.05).The apoptosis index of HUVEC were higher compared with the control groups(P0.05).The percent of the cultured cells in G1 phase in all high glucose groups were increased, the percent in S phase were largely decreased(P0.05). Conclusion Exposed to high glucose,the apoptosis of HUVEC was accelerated and the suppressive effect of high glucose on the proliferation of HUVEC was observed.Endothelial cells were possibly arrested in G1/S checkpoint.
Proliferation index
Thymidine
Cite
Citations (0)
Objective To observe proliferative effect of keratinocyte growth factor (KGF) on 1C6 cells and 4C18 cells originated from thymic epithelial cell. Methods 1C6 cells and 4C18 cells originated from the medulla and cortex of mouse thymic epithelial,and RAW cells from mouse peritoneal macrophage were cultured in complete culture medium with different concentrations (25,50,100,200,400,800 ng/ml) of KGF and without KGF (control group) for 24,48 and 72 h respectively,and cell proliferation rate was analyzed by MTT assay,at the same time proliferation curves were made. Results MTT assay showed that when KGF concentration was less than 200 ng/ml,the proliferation rate of 1C6 cells increased as the concentration raising,when KGF concentration was 200 ng/ml,the proliferation rate of 1C6 cells reached the peak value,and when KGF concentration was more than 200 ng/ml,the proliferation rate of 1C6 cells decreased gradually as the concentration declining. Compared with the control group respectively,the proliferative effects on 1C6 cells affected by concentrations of 100,200,400 ng/ml of KGF for 24,48 and 72 h were obvious (all P 0.05). When KGF concentration was less than 100 ng/ml,the proliferation rate of 4C18 cells increased as the concentration raising,when KGF concentration was 100 ng/ml,the proliferation rate of 4C18 cells reached the peak value,and when KGF concentration was more than 100 ng/ml,the proliferation rate of 4C18 cells decreased gradually as the concentration declining. Compared with the control group respectively,the proliferative effects on 4C18 cells affected by concentrations of 50,100,200 and 400 ng/ml of KGF for 24,48 and 72 h were obvious (all P 0.05). But different concentration of KGF had no proliferation effect on RAW cells. Conclusion KGF could promote the proliferation of 1C6 cells and 4C18 cells originated from thymic epithelial cells.
Keratinocyte growth factor
MTT assay
Cell counting
Cite
Citations (0)
AIM: To evaluate the effect of hepatocyte growth factor (HGF) on the proliferation and apoptosis in HepG2 cells. METHODS: HepG2 cells were cultured for 2, 4, 6 d in the presence of HGF at different concentrations of 0, 1, 10, 50 μg/L. The effects of HGF on the proliferation of HepG2 cells was evaluated by MTT assay. The flow cytometry (FCM) was used to detect phase distribution of the cycles and apoptosis rate. Expression of proliferating cell nuclear antigen (PCNA) was detected with immunocytochemical technique. RESULTS: HGF inhibited the cell proliferation and induced the apoptosis in HepG2 in a time- and concentration-dependent manner. FCM also showed a sub-G1 cell apoptotic peak and a significant increace of apoptosis rate induced by HGF(P0.05). At the concentration of 50 μg/L, HGF treatments for 4 and 6 d depressed the proliferation of HepG2 cells by (30.7±10.2)% and (49.8 ±11.1)% , while at 1 μg/L, HGF decreased the proliferation by (10.7±11.2)% and (4.2±9.4)%. Differences between the 2 groups were statistically significant (P0.05, P0.01). DNA ploidy analysis showed that S phase cells were significantly decreased and quiescent G0/G1 phase cells were accumulated by raising HGF concentration. Percentages of S phase cells in 10 and 50 μg/L groups were (7.5±4.4)%, (7.8±1.6)%, significantly lower than the control(16.6 ±2.8) %; percentages of G0/G1 phase cells were (80.5±3.2)% and (73.1±3.9)%, significantly higher than the control (59.6±6.2)% (P0.05, P 0.01). Levels of PCNA were obviously down-regulated in HGF group compared with control group. CONCLUSION: HGF significantly inhibits the proliferation and induces the apoptosis in HepG2, which may be involved in G0/G1 phase arrest.
MTT assay
Cite
Citations (0)
Objective To investigate the effect of recombinant human interleukin-10(IL-10)on proliferation of adventitial fibroblasts and expression of syndecan-4 protein induced by tumor necrosis factor-α(TNF-α).Methods NIH 3T3 cells were cultured in vitro and exposed to 20 ng/mL TNF-α,100 ng/mL IL-10,200 ng/mL IL-10,100 ng/mL IL-10 with 20 ng/mL TNF-α,and 200 ng/mL IL-10 with 20 ng/mL TNF-α respectively.All groups were treated for 24 h in vitro,and were compared with the control group.The ratio of proliferation of NIH/3T3 cells was determined by non-radioactive MTS/PMS assay and the expression of syndecan-4 protein was evaluated by Western blotting using anti-syndecan-4 antibodies.Results Statistical analysis showed that,(1) Compared to the control group,TNF-α significantly stimulated the proliferation of NIH/3T3 cells at the concentration of 20 ng/mL in TNF-α group(P0.001).IL-10 alone had no effect on NIH/3T3 cells growth(P0.05),but significantly inhibited NIH/3T3 cells proliferation induced by TNF-α(P0.01).(2) Compared with the control group,the expression of syndecan-4 protein in NIH/3T3 cells was significantly enhanced by TNF-α at the concentration of 20 ng/mL(P0.05).IL-10 alone had no effect on the expression of syndecan-4 protein(P0.05),but significantly inhibited the expression of syndecan-4 induced by TNF-α(P0.001).Conclusion IL-10 could inhibit expression of syndean-4 protein in NIH/3T3 cells induced by TNF-α in vitro.
Syndecan 1
3T3 cells
Cite
Citations (0)
Objective To investigate the effects of β-sitosterol on the proliferation of NK cells and human pancreatic cancer cell line SW-1990,and to discuss the possible mechanism of action.Methods The NK cells were cultured in special culture established by our laboratory.After culturing the NK cells and SW-1990 cell line with different concentrations of β-sitosterol,the effect of β-sitosterol on growth of NK cells and SW-1990 cell line was detected by MTT assay,cell cycle and apoptosis of SW-1990 cell line were analyzed by Flow Cytometry(FCM).In addition,the cytotoxic activity of NK cells in SW-1990 cell line induced by β-sitosterol in different concentrations was evaluated with LDH method.Results After culturing with 1.25 to 10 μg/mL of β-sitosterol for 48 hours,the growth inhibition rate of NK cells was minus,decreased significantly compared with the control group(P0.05),when the concentration of β-sitosterol was increased to 40-80 μg/mL,the growth inhibition rate of NK cells increased significantly compared with the control group(P0.05).10~80 μg/mL of β-sitosterol can significant inhibit the growth of SW-1990(P0.05),apoptosis rate of SW-1990 cell line was 27.18% when β-sitosterol was 10 μg/mL,increased significantly compared with the control group and the low concentration group(P0.05).Cell cycle of SW-1990 was arrested in S-phase induced by β-sitosterol.After culturing with 1.25 and 2.5 μg/mL of β-sitosterol for 48 hours,the cytotoxic activity of NK cells on SW-1990 increased significantly when compared with the control group(P0.05),and reached a peak(76.3%) in 1.25 μg/mL.When the concentration of β-sitosterol above 5 μg/mL,the cytotoxic activity of NK cells decreased with increasing drug concentration.Conclusion The β-sitosterol could strengthen the cytotoxicity of NK cells against human pancreatic cancer cells SW-1990.
Cite
Citations (0)
To determine the effects of nerve growth factor (NGF) on proliferation of hepatic stellate cells (HSCs) and investigate the related molecular mechanism.After incubating cultured HSCs for 24 h with different concentrations of NGF (100, 200 or 400 ng/mL), the cell proliferation was observed by XTT colorimetric assay and cell cycle was detected by flow cytometry. Morphological changes in response to a 24 h exposure to 100 ng/mL NGF were observed by transmission electron microscopy.NGF significantly inhibited HSC proliferation (P less than 0.05) in a dose-independent manner. The optical densities of the XTT colorimetric assay were 0.66+/-0.03 for 100 ng/mL NGF, 0.69+/-0.03 for 200 ng/mL NGF, and 0.66+/-0.03 for 400 ng/mL NGF, all of which were significantly lower than that of the control group (0.73+/-0.01; P less than 0.05). All concentrations of NGF led to significantly higher numbers of HSCs in the G2 phase (100 ng/mL: 14.83+/-5.41%, 200 ng/mL: 14.73+/-2.50%, and 400 ng/mL: 14.87+/-2.06%), compared to that detected in the control group (7.47+/-4.39%; P less than 0.05). Twenty-four hours of exposure to 100 ng/mL NGF caused morphological changes indicative of apoptosis.NGF inhibits the proliferation of HSCs, possibly by arresting the cells in the G2 phase of the cell cycle. NGF-inhibited cells may also undergo apoptosis.
Hepatic stellate cell
Cite
Citations (0)