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Objective To investigate the biological features of epidermal growth factor receptor inhibitor AG1478 on cervical carcinoma cell. Methods The proliferation of HeLa cells under AG1478 stimulation was determined by CCK8 assay. The expression of EGFR downstream signaling protein and apoptotic relative protein were examined by Western blot and transcription of apotosis?related genes were measured by RT?qPCR in AG 1478 treated HeLa cells. Nuclear transport of phosphorylated ERK were measured through ICC assay. TUNEL assay was used to determine early stage of apoptosis. Results CCK8 assay showed that AG1478 inhibit the proliferation of HeLa cells and also block phosphorylated level of EGFR ,ERK and AKT. Furthermore ,nuclear transport of phosphorylated ERK upon EGF stimulation were blocked and pro?apoptotic proteins were up?regluated with activat ed cleaved Caspases. Conclusion AG1478 inhibited the proliferation and induced apoptosis in HeLa cells and it could be a potential therapeutic drug for cervical carcinoma.
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AIM:To explore the role of Bcl-2 and PCNA expression in the injury of rat myoblasts induced by hydrogen peroxide (H2O2).METHODS:Rat myoblasts at growth phase were divided into 4 groups based on basic fibroblast growth factor (bFGF) and H2O2 levels:normal control group, bFGF group, model group (H2O2 group) and treatment group (bFGF+H2O2 group).The expression of Bcl-2 and Bax was observed by immunohistochemistry and fluorescence methods.The protein levels of Bax, Bcl-2 and PCNA were determined by Western blot.RESULTS:Compared with model group, both immunofuorescence and fluorescence in treatment group showed enhanced Bcl-2 and low expression of Bax.Furthermore, the results of Western blot showed up-regulated PCNA and Bcl-2 protein and decreased Bax expression in treatment group.CONCLUSION:Oxidative stress results in the pathologic changes of myoblasts, and the up-regulation of Bcl-2 and PCNA may attenuate myoblast injury.
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<p><b>OBJECTIVE</b>To investigate the effect of exogenous recombinant human basic fibroblast growth factor (rhbFGF) on the healing of muscles in rats after deep tissue injury of pressure ulcers.</p><p><b>METHODS</b>Forty-eight SD rats were randomly divided into normal control group, injury control group, post injury day (PID) 4 group, PID 7 group, PID 14 group, PID 21 group according to the random number table, with 8 rats in each group. The rats in normal control group did not receive any treatment, whereas the rats in the latter 5 groups were established the deep tissue injury of pressure ulcer model on both sides of the gracilis muscle on the hind limb. The rats in injury control group did not receive any treatment after injury, while the rats in the latter 4 groups were given subcutaneous injection of 0.1 mL rhbFGF to the left gracilis in a dosage of 100 µg/mL immediately after injury, and an equal volume of normal saline (NS) was injected to right gracilis, once every other day. The rats in injury control group were sacrificed immediately after injury, and the rats in normal control group were sacrificed at the same time point. The rats in the other 4 groups were sacrificed on PID 4, 7, 14, 21, and the gracilis muscles on both sides were harvested respectively. The morphology of the gracilis muscle was examined after HE staining. The expression of myogenin in the tissues was detected by immunofluorescence method. The levels of muscle structural proteins myosin heavy chain (MyHC), phosphorylated protein kinase B (Akt), and phosphorylated mammalian target of rapamycin (mTOR) were determined by Western blotting. Data were processed with one-way analysis of variance and LSD test.</p><p><b>RESULTS</b>(1) In normal control group, the nuclei of graciles cells were in uniform size, and they were closely arranged with clear structure, and there were no significant infiltration of inflammatory cells. In injury control group, the nuclei of graciles cells showed signs of pyknosis, dissolution, fracture and structural disorder. Swelling of muscle cells, inflammation infiltration, structural disorder and other pathological signs of injury phenomena in graciles of PID 4 group, PID 7 group, PID 14 group, PID 21 group after rhbFGF treatment were milder compared with those after NS treatment. In addition, the numbers of regenerated myocytes in graciles of PID 4 group, PID 7 group, PID 14 group, PID 21 group after rhbFGF treatment were higher than those after NS treatment. (2) The numbers of graciles myogenin positive cells in normal control group and injury control group were respectively 28 ± 17 and 42 ± 28. The numbers of graciles myogenin positive cells in PID 4 group, PID 7 group, PID 14 group after NS treatment were 100 ± 50, 196 ± 87, 460 ± 110 respectively, while the numbers of graciles myogenin positive cells in PID 4 group, PID 7 group, PID 14 group after rhbFGF treatment were 174 ± 34, 717 ± 182, 613 ± 122 respectively, and the numbers of graciles myogenin positive cells after rhbFGF treatment were significantly higher than those after NS treatment in each group(P < 0.05 or P < 0.01). The number of graciles myogenin positive cells in PID 21 group after rhbFGF treatment was 109 ± 34, which was significantly lower than that after NS treatment (218 ± 71, P < 0.05). (3) The expression of MyHC in graciles in normal control group was high, which was decreased in injury control group. Both the expressions of MyHC in graciles in PID 4 group, PID 7 group, PID 14 group, PID 21 group after treatment of NS and rhbFGF showed a trend of gradual elevation, while the expressions of MyHC in graciles after rhbFGF treatment were significantly higher than those after NS treatment (P < 0.05 or P < 0.01). The expression of MyHC in graciles in PID 21 group showed a high level, and it was similar to that of the normal control group (P > 0.05). The expressions of phosphorylated Akt and phosphorylated mTOR in graciles of normal control group were low, and the expression of phosphorylated Akt in graciles increased in injury control group, while the expression of phosphorylated mTOR in graciles decreased in injury control group. The expressions of phosphorylated Akt and phosphorylated mTOR in graciles of PID 4 group, PID 7 group, PID 14 group, PID 21 group after treatment with rhbFGF showed a trend of elevation in the beginning but declined afterwards. The expressions of phosphorylated Akt and phosphorylated mTOR in graciles of PID 4 group after rhbFGF treatment were significantly lower than those after NS treatment (P <0 .05 or P < 0.01). The expressions of phosphorylated Akt and phosphorylated mTOR in graciles of PID 7 group, PID 14 group, PID 21 group after rhbFGF treatment were significantly higher than those after NS treatment (P < 0.05 or P < 0.01).</p><p><b>CONCLUSIONS</b>Exogenous rhbFGF may effectively facilitate the healing of muscle structure and accelerate the regeneration of muscles in rats after deep tissue injury of pressure ulcers, and its mechanism may be related to the improvement of the expression of myogenin and enhancement of the expression of protein of muscle growth-related signaling pathways.</p>