To investigate whether the expression of exogenous heme oxygenase-1 (HO-1) gene within vascular smooth muscle cells (VSMC) could protect the cells from free radical attack and inhibit cell proliferation, we establishe...To investigate whether the expression of exogenous heme oxygenase-1 (HO-1) gene within vascular smooth muscle cells (VSMC) could protect the cells from free radical attack and inhibit cell proliferation, we established an in vitro transfection of human HO-1 gene into rat VSMC mediated by a retroviral vector. The results showed that the profound expression of HO-1 protein as well as HO activity was 1.8- and 2.0-fold increased respectively in the transfected cells compared to the non-transfected ones. The treatment of VSMC with different concentrations of H2O2 led to the remarkable cell damage as indicated by survival rate and LDH leakage. However, the resistance of the HO-1 transfected VSMC against H2O2 was significantly raised. This protective effect was dramatically diminished when the transfected VSMC were pretreated with ZnPP-IX, a specific inhibitor of HO, for 24 h. In addition, we found that the growth potential of the transfected cells was significantly inhibited directly by increased activity of HO-1, and this effect might be related to decreased phosphorylation of MAPK. These results suggest that the overexpression of introduced hHO-1 is potentially able to reduce the risk factors of atherosclerosis, partially due to its cellular protection against oxidative injury and to its inhibitory effect on cellular proliferation.展开更多
AIM: To observe the growth suppression effect of exogenous introduction of early growth response gene-1 (Egr-1 gene) on esophageal carcinoma tissue as well as on esophageal carcinoma cell line Eca109 and to explore th...AIM: To observe the growth suppression effect of exogenous introduction of early growth response gene-1 (Egr-1 gene) on esophageal carcinoma tissue as well as on esophageal carcinoma cell line Eca109 and to explore the potential application of Egr-1 gene in gene therapy of tumor. METHODS: Eukaryotic expression vector of PCMV-Egr-1 plasmid was introduced into Eca109 cell line which expressed no Egr-1 protein originally with lipofectamine transfection method. The introduction and expression of PCMV-Egr-1 plasmid into Eca109 cell line was confirmed by G418 selection culture, PCR amplification of neogene contained in the vector, Western blot analysis and immunocytochemical analysis. The cell growth curve, soft agar colony formation rate and tumorigenicity in SCID mice were examined to demonstrate the growth suppression effect of exogenous Egr-1 gene on Eca109 cell line. The Egr-1 mRNA and Egr-1 protein were also detected in 50 surgical specimens of esophageal carcinoma by in situ hybridization and immunohistochemistry. RESULTS: Exogenous Egr-1 gene was introduced successfully into Eca109 cell line and expressed Egr-1 protein stably. The transfected Eca109 cell line grew more slowly than control Eca109 as shown by cell growth curves, the soft agar colony formation rate (4.0% vs 6.9%, P 【 0.01) and the average growth rate of tumor in SCID mice (35.5 +/- 7.6 vs 65.8 +/- 7.6, P 【 0.05). The expression level of Egr-1 mRNA and protein significantly increased in dysplastic epithelia adjacent to cancer rather than in cancer tissues (65.8% vs 20.0% by ISH and 57.9% vs 0.01). CONCLUSION: Exogenous Egr-1 gene shows the strong effect of growth inhibition in Eca109 cell line. Egr-1 in the cancer tissue shows down-regulated expression that supports the inhibited function of Egr-1 in cancer growth and suggests Egr-1 may have an important role in gene therapy of esophageal carcinoma.展开更多
Based on the subdivision into three members of the Lower Cretaceous Jiufotang Formation in western Liaoning, this paper deals mainly with the division and correlation of precious fossil birdand reptile-bearing beds of...Based on the subdivision into three members of the Lower Cretaceous Jiufotang Formation in western Liaoning, this paper deals mainly with the division and correlation of precious fossil birdand reptile-bearing beds of the formation in the Dachengzi, Chaoyang, Dapingfang.Meileyingzi and Fuxin-Yixian basins. Among them, the precious fossil-bearing beds in the Dachengzi Basin may be recognized as the Xidagou Bed of the second member and the Yangcaogoudonggou Bed of the third member; those in the Chaoyang Basin may be confirmed as the Shangheshou Bed of the second member and the Dongpochi Bed of the third member; those in the Dapingfang-Meileyingzi Basin are as the Lamagou Bed of the second member, the Huanghuagou Bed of the lower third member and the Yuanjiawa Bed of the upper third member; and those in the Fuxin-Yixian Basin are listed as the Tuanshanzi Bed of the second member and the Pijiagou Bed of the third member. Since these basins are distinctly separated and the bird and reptile fossils are mostly new genera and species, we have to use the associated fossil ostracod assemblages as index to correlate the Xidagou Bed with the Shangheshou Bed and the Lamagou Bed, and to correlate the Yuanjiawa Bed with the Yangcaogoudonggou Bed and the Pijiagou Bed. Primarily, we established the sequence of the precious fossil-bearing beds of the Jiufotang Formation in western Liaoning. They are represented by, in ascending order, the Xidagou Bed of the second member, the Huanghuagou Bed of the lower third member, and the Yuanjiawa Bed of the upper third member. Obviously, this work has significance for the study on the time-space distribution and radiation of birds and dinosaurs during the period of the middle Early Cretaceous.展开更多
基金This work was kindly supported by Na-tional Natural Science Foundation of China(No.39670308)
文摘To investigate whether the expression of exogenous heme oxygenase-1 (HO-1) gene within vascular smooth muscle cells (VSMC) could protect the cells from free radical attack and inhibit cell proliferation, we established an in vitro transfection of human HO-1 gene into rat VSMC mediated by a retroviral vector. The results showed that the profound expression of HO-1 protein as well as HO activity was 1.8- and 2.0-fold increased respectively in the transfected cells compared to the non-transfected ones. The treatment of VSMC with different concentrations of H2O2 led to the remarkable cell damage as indicated by survival rate and LDH leakage. However, the resistance of the HO-1 transfected VSMC against H2O2 was significantly raised. This protective effect was dramatically diminished when the transfected VSMC were pretreated with ZnPP-IX, a specific inhibitor of HO, for 24 h. In addition, we found that the growth potential of the transfected cells was significantly inhibited directly by increased activity of HO-1, and this effect might be related to decreased phosphorylation of MAPK. These results suggest that the overexpression of introduced hHO-1 is potentially able to reduce the risk factors of atherosclerosis, partially due to its cellular protection against oxidative injury and to its inhibitory effect on cellular proliferation.
基金Supported by the National Natural Science Foundation of China,No.39670298.
文摘AIM: To observe the growth suppression effect of exogenous introduction of early growth response gene-1 (Egr-1 gene) on esophageal carcinoma tissue as well as on esophageal carcinoma cell line Eca109 and to explore the potential application of Egr-1 gene in gene therapy of tumor. METHODS: Eukaryotic expression vector of PCMV-Egr-1 plasmid was introduced into Eca109 cell line which expressed no Egr-1 protein originally with lipofectamine transfection method. The introduction and expression of PCMV-Egr-1 plasmid into Eca109 cell line was confirmed by G418 selection culture, PCR amplification of neogene contained in the vector, Western blot analysis and immunocytochemical analysis. The cell growth curve, soft agar colony formation rate and tumorigenicity in SCID mice were examined to demonstrate the growth suppression effect of exogenous Egr-1 gene on Eca109 cell line. The Egr-1 mRNA and Egr-1 protein were also detected in 50 surgical specimens of esophageal carcinoma by in situ hybridization and immunohistochemistry. RESULTS: Exogenous Egr-1 gene was introduced successfully into Eca109 cell line and expressed Egr-1 protein stably. The transfected Eca109 cell line grew more slowly than control Eca109 as shown by cell growth curves, the soft agar colony formation rate (4.0% vs 6.9%, P 【 0.01) and the average growth rate of tumor in SCID mice (35.5 +/- 7.6 vs 65.8 +/- 7.6, P 【 0.05). The expression level of Egr-1 mRNA and protein significantly increased in dysplastic epithelia adjacent to cancer rather than in cancer tissues (65.8% vs 20.0% by ISH and 57.9% vs 0.01). CONCLUSION: Exogenous Egr-1 gene shows the strong effect of growth inhibition in Eca109 cell line. Egr-1 in the cancer tissue shows down-regulated expression that supports the inhibited function of Egr-1 in cancer growth and suggests Egr-1 may have an important role in gene therapy of esophageal carcinoma.
文摘Based on the subdivision into three members of the Lower Cretaceous Jiufotang Formation in western Liaoning, this paper deals mainly with the division and correlation of precious fossil birdand reptile-bearing beds of the formation in the Dachengzi, Chaoyang, Dapingfang.Meileyingzi and Fuxin-Yixian basins. Among them, the precious fossil-bearing beds in the Dachengzi Basin may be recognized as the Xidagou Bed of the second member and the Yangcaogoudonggou Bed of the third member; those in the Chaoyang Basin may be confirmed as the Shangheshou Bed of the second member and the Dongpochi Bed of the third member; those in the Dapingfang-Meileyingzi Basin are as the Lamagou Bed of the second member, the Huanghuagou Bed of the lower third member and the Yuanjiawa Bed of the upper third member; and those in the Fuxin-Yixian Basin are listed as the Tuanshanzi Bed of the second member and the Pijiagou Bed of the third member. Since these basins are distinctly separated and the bird and reptile fossils are mostly new genera and species, we have to use the associated fossil ostracod assemblages as index to correlate the Xidagou Bed with the Shangheshou Bed and the Lamagou Bed, and to correlate the Yuanjiawa Bed with the Yangcaogoudonggou Bed and the Pijiagou Bed. Primarily, we established the sequence of the precious fossil-bearing beds of the Jiufotang Formation in western Liaoning. They are represented by, in ascending order, the Xidagou Bed of the second member, the Huanghuagou Bed of the lower third member, and the Yuanjiawa Bed of the upper third member. Obviously, this work has significance for the study on the time-space distribution and radiation of birds and dinosaurs during the period of the middle Early Cretaceous.
