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Objective To investigate the effect of hepatocyte growth factor (HGF) on migration and apoptosis of,as well as phosphorylated-Akt (p-Akt) expression in cultured human eccrine sweat gland epithehal cells (hESGc).Methods The first generation of hESGc were cultured in keratinocyte serum free medium (KSFM) and treated with various concentrations (2,20,40μg/L) of HGF for different durations.Then,cell scratch test was performed to detect cell migration,a double staining flow cytometry assay using annexin VFITC/propidium iodide to detect cell apoptosis.and Western blot to measure the expression of p-Akt.Results HGF of 2μg/L had no effect on the migration of hESGc,while that of 20 μg/L and 40μg/L could promote the migration of hESGc by 33.2% and 228.2%.respectively.The average number of cells migrating into the scrach zone was significantly lower in untreated cell group than that in 20 and 40μg/L HGF-treated cell group (17.3±5.5 vs 23.0±6.3 and 56.7±7.9,t=2.653, 15.858,P<0.05,0.01, respectively).The apoptosis rate was 14.76% in untreated cells,14.16%,13.5% and 8.87% in cells treated with HGF of 2,20 and 40μ/L, respectively;there was a significant difference between untreated cells and 40μg/L HGF-treated cells (t=7.852,P<0.01).HGF could activate the phosphorylation of Akt protein and increase the expression of p-Akt.Conclusion HGF could promote the migration of,inhibit the apoptosis of,and stimulate the p-Akt expression in.hESGc.
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[Objective] To explore the cytoprotecfion of hydrogen sulfide (H_2S) against cobalt chloride (CoCl_2)-induced apeptosis in PC12 cells and the underlying mechanisms. [Methods] CoCl_2 (a chemical hypoxia-mimetic agent) was used to establish the chemical hypoxia-induced PC12 cell injuries model. Sodium hydrosulfide (NaHS) was used as a H_2S donor. The viability of PC12 cells was measured by CCK-8 assay. The percentage of apeptotic cells was assessed by propidium iodide stain flow cytometry (FCM). The morphological change of apeptotic cells was tested by using the chromatin dye Hoechst 33258. The mitochondrial membrane potential (MMP) was analyzed by rhodamine 123 staining and photofluorography. The level of reactive oxygen species (ROS) in PC12 cells was measured by DCFH-DA staining and photofluorography. [Results] CoCl_2 induced a decrease in cell viability and an increase in percentage of apeptosis in PC12 cells along with dissipation of MMP as well as overproduction of ROS. When PC12 cells were treated with Naris 30 min before CoCl_2 treatment a decrease in viability of PC12 cells induced by 600 μmol/L CoCl_2 was concentration-dependently blocked by NaHs (100, 200, and 400 μmol/L). Pretreatment with NaHS at 200 and 400 μmol/L obviously reduced the apepetotic percentage of PC12 cells induced by 600 μmol/L CoCl_2 and inhibited the dissipation of MMP and overproduction of ROS. [Conclusion] H_2S protected PC12 cells against CoCl_2-induced apeptosis, which may be associated with the inhibition of H_2S on the dissipation of MMP and overproduction of ROS induced by CoCl_2.
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Objective To investigate the effect of zerumbone on the proliferation and apoptosis of human pancreatic cancer cell line PANC1 and its possible mechanism.Methods Zerumbone of various concentrations (3.75, 7.5, 15, 30, 60 μg/ml) were used to treat PANC1, and cells without treatment were used as control.CCK-8 assay was used to detect the inhibitory rate of cell proliferation.Cell apoptosis analysis was determined by using Hoechst 33342 staining and flow cytometry.Western blotting was performed to evaluate the phosphorylation Statl ( p-STAT1 ), and Bax and Bcl2 protein expression.Results Zerumbone caused a time- and dose-dependent reduction of cell viability in PANC1 cells.After 48h treatment of Zerumbone of 15 μg/ml, cells inhibitory rate was increased to (72.8 ± 2.72 )%, and classic apoptosis morphology was observed, with apoptosis rate was 14.2%.At the same time, p-STAT1, and Bax protein expression was significantly increased (0.654 ±0.048 vs 0.074 ±0.011, 0.577 ±0.044 vs 0.218 ±0.027,P＜0.05);Bcl-2 protein expression was significantly decreased (0.162 ± 0.029 vs 0.459 ± 0.034, P＜0.05).Conclusions Zerumbone may inhibit the proliferation of PANC1 cells and inducing cell apoptosis,which may be related to the up-regulation of STAT1's activity and Bcl-2/Bax ratio.
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Objective To study the influences by a Cyelin-dependent kinase inhibitor Roscovitine on cell cycle in mitotic hepatoma carcinoma cell SMMC-7721. Methods Microscope,MTT, flow cytometry and R-T PCR were used to observe the effects of Roscovitine on morphology, proliferation, cell cycle, apoptosis and the mRNA expression of CDK2, Caspase-3, bcl-2 in SMMC-7721 cells. Results Roscovitine inhibited the proliferation of SMMC-7721 cells in dosage and time dependent manner and induced apoptosis. Flow cytometry showed the ratio of G0, G1 increased. R-T PCR showed that the expression of bcl-2 reduced, Caspase-3 increased. Conclusion Reseovitine can inhibit the growth, proliferation, block the cell cycle at G0/G1 and promotes apoptosis of mitotic SMMC-7721 cells, and the mechanism of apoptosis is dependent on the activity of bcl-2 and Caspase-3.
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Objective To explore the effect of sodium fluoride on DNA damage and apoptosis on ox peripheral blood lymphocyte cultured in vitro.Methods Using lymphoeytes separation medium lymphocytes were separated and different concentration of NaF(0,4,8,12,16 mg/L)were added into the cultual system of lymphocytes for 24 h.Cell viability was measured by MTT.nuclear changes stained by acridine orange-ethidium bromine staining (AO/EB)were observed under fluoroscope,DNA fragment was measured by agarose gel electrophoresis.DNA damage was detected by alkaline SCGE.The differences between each groups were compared.Results Cells were exposed to 4,8,16 and 24 mg/L NaF in 24 h,cell survival rates,respectively being(73.743±0.552)%,(69.184±0.724)%,(65.736±0.055)%and(63.651±0.287)%,decreased significantly compared to control group.There were distinctive cell apoptosis,evident DNA damage and visible dose-effect relation(R2=0.7456).Conclusions A certain concentration of sodium fluoride lcads to lymphocytes apoptosis and DNA damage.