Transcriptional Factor Snail Mediates Epithelial-Mesenchymal Transition in Human Bronchial Epithelial Cells Induced by Silica / 生物医学与环境科学（英文）

Epithelial-mesenchymal transition (EMT) plays an important role in fibrotic diseases. We have previously showed that silica induces EMT in human bronchial epithelial cells (BECs); however, the underlying mechanism of silica-induced EMT is poorly understood. In the present study, we investigated the role of Snail in silica-induced EMT in human BECs in vitro. Human BECs were treated with silica at various concentrations and incubation times. Then MTT assay, western blot, electrophoretic mobility shift assay (EMSA), and small interfering RNA (siRNA) transfection were performed. We found that silica increased the expression and DNA binding activity of Snail in human BECs. SNAI siRNA inhibited the silica-induced expression of Snail. Moreover, SNAI siRNA upregulated the expression of epithelial marker E-cadherin, but attenuated the expression of mesenchymal marker α-smooth muscle actin and vimentin in silica-stimulated cells. These results suggest that Snail mediates the silica-induced EMT in human BECs.

Roles of Rho/Rock signaling pathway in silica-induced epithelial-mesenchymal transition in human bronchial epithelial cells / 生物医学与环境科学（英文）

<p><b>OBJECTIVE</b>To investigate the roles of Rho/Rock signaling pathway in silica-induced Epithelial-mesenchymal transition (EMT) in human bronchial epithelial cells (BEC) in vitro.</p><p><b>METHODS</b>Human BEC were incubated with silica with various concentrations for indicated times. Cell viability was assayed by MTT test. Morphologic Changes were observed by microscope. Mesenchymal marker α-smooth muscle actin (α-SMA), vimentin (Vim), and epithelial marker E-cadherin (E-cad) were analyzed by Western Blot. The pull-down assay was used to measure Rho activity. In the prevention experiments, the specific inhibitor for Rho effector ROCK (Y27632) was used to inhibit the activity of Rho.</p><p><b>RESULTS</b>Human BEC stimulated with silica were converted from a "cobblestone" epithelial structure into an elongated fibroblast-like shape structure. Incubation of human BEC with silica induced de novo expression of α-SMA and Vim, and loss of E-cad. Also, silica treatment resulted in Rho activation in human BEC. Y27632 up-regulated the E-cad expression but attenuated α-SMA and Vim expression in silica-stimulated cells.</p><p><b>CONCLUSION</b>The activation of Rho/ROCK signaling pathways is most likely involved in Silica-induced EMT in human bronchial epithelial cells.</p>

Changes of neuroglobin in the pathologic process of contusion and laceration of brain in children / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To study the role of neuroglobin (Ngb) in the pathologic process of contusion and laceration of brain in children.</p><p><b>METHODS</b>The proteins in the brain tissue were extracted by two-dimensional gel electrophoresis in 3 children undergoing brain ventricular neoplasms resection (normal brain tissue) and in 8 children with contusion and laceration of brain. The image analysis was done using the PDQuest 7.0 software. The differential protein spots were detected and analyzed with Applied Biosystems Voyager System 4307 MALDI-TOF Mass Spectrometer and bioinformatical skills. Ngb expression in the brain tissue was measured using immunohistochemisty. Ngb expression in plasma was measured using ELISA in 15 children with contusion and laceration of brain and 10 healthy children.</p><p><b>RESULTS</b>Expression maps of the brain tissue were established by two-dimensional gel electrophoresis in children with contusion and laceration of brain and healthy children. Six differential protein spots were found and 5 of them were identified by mass spectrum. Immunohistochemisty assay showed that Ngb expression in the brain tissue in children with contusion and laceration of brain was significantly higher than in normal controls (P<0.05). ELISA results showed that Ngb expression in the plasma increased significantly 6, 12, 18, 24 and 48 hours after trauma in children with contusion and laceration of brain compared with healthy children (P<0.01).</p><p><b>CONCLUSIONS</b>Ngb may play an important role in the pathologic process of contusion and laceration of brain in children.</p>

Expression of EVEC in ovarian carcinoma and its biological significance / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To investigate the expression of EVEC in ovarian carcinoma and explore its biological significance.</p><p><b>METHODS</b>The expression of EVEC in 22 specimens of normal ovarian tissues and 63 specimens of ovarian cancers was detected by RT-PCR and Western blotting analysis, respectively.</p><p><b>RESULTS</b>RT-PCR showed that the expression level of EVEC in stage I-II ovarian cancer (0.199 ± 0.014) was significantly higher than that in stage III-IV ovarian cancer (0.155 ± 0.015, P < 0.05), and significantly lower than that in normal ovarian tissues (0.415 ± 0.055, P < 0.05). There was no significant difference between the expression levels of EVEC in primary sites and that in corresponding metastatic sites of ovarian cancer (P > 0.05). Furthermore, the results of Western blot also showed that the protein expression level of EVEC in stage I-II ovarian cancer was also significantly lower than that in normal ovarian tissues (0.179 ± 0.026 vs. 0.543 ± 0.032, P < 0.05), and higher than that in stage III-IV ovarian cancer (0.179 ± 0.026 vs. 0.115 ± 0.023, P < 0.05). The EVEC expression level in the epiploic metastasis of stage I-II ovarian cancer was significantly higher than that of stage III-IV ovarian cancer (0.201 ± 0.028 vs. 0.101 ± 0.037, P < 0.05). The expression of EVEC in ovarian carcinoma had no correlation with age, pathologic classification and histological grade (P > 0.05).</p><p><b>CONCLUSIONS</b>EVEC is closely related with carcinoma metastasis. The expression of EVEC in ovarian cancer and its metastatic sites was remarkably decreased. EVEC may play a negative role in the development and metastasis of ovarian cancer and may be a valuable marker in estimation of the prognosis for patients.</p>

Effect of FGF-2 on survivin and subcellular location of Smac in human small cell lung cancer cell NCI-H446 / 中南大学学报(医学版)

OBJECTIVE@#To investigate the effect of basic fibroblast growth factor (FGF-2)on survivin and subcellular location of Smac in human small cell lung cancer (SCLC) cell NCI-H446.@*METHODS@#Western blot was used to detect the expression of survivin protein induced by FGF-2. The release of Smac from mitochondria to cytoplasm affected by FGF-2 was observed by Western blot and immunofluorescence. Apoptosis of NCI-H446 cells was detected with flow cytometry and Hoechst 33258 staining.@*RESULTS@#The expression of survivin could be up-regulated in response to FGF-2 treatment in NCI-H446 cells, and the level of survivin expression is related to the concentration and time of FGF-2 treatment. FGF-2 could inhibit the release of Smac from the mitochondria to cytoplasm induced by serum starving. FGF-2 could inhibit the apoptosis induced by serum starving.@*CONCLUSION@#FGF-2 up-regulates the expression of survivin protein in NCI-H446 cells, and blocks the release of Smac from mitochondria cytoplasm. Survivin and Smac might play important roles in the apoptosis inhibited by FGF-2.

Expression and significance of Notch1, Jagged1 and VEGF in human non-small cell lung cancer / 中南大学学报(医学版)

OBJECTIVE@#To determine the expressions of Notch1, Jagged1 and vascular endothelial growth factor (VEGF) in human non-small cell lung cancer (NSCLC) and to explore its clinical pathological significance.@*METHODS@#Immunohistochemical SP method was used to detect the expressions of Notch1, Jagged1 and VEGF in 65 patients with NSCLC and 15 normal epithelial tissues of the lung, and the relationship between them and clinic-pathological parameters were analyzed.@*RESULTS@#The positive rates of Notch1, Jagged1 and VEGF in NSCLC were 81.5%, 83.1% and 93.8%, respectively, higher than those in normal epithelial tissues of the lung (P<0.05). The positive expression levels of Notch1 and VEGF were closely associated with the tumor stage and the lymph node metastasis (P<0.05). The positive expression levels of Jagged1 was positively correlated with the pathological type and lymph node metastasis. There was a positive correlation between Notch1, Jagged1 and VEGF.@*CONCLUSION@#Notch1, Jagged1 and VEGF protein may play an important role in the pathway of carcinogenesis and progression of NSCLC. The up-regulation of Notch1, Jagged1 and VEGF protein expression probably predict NSCLC carrying relatively strong permeation and metastasis.

Expression of survivin and proliferating cell nuclear antigen in human non-small cell lung cancer / 中南大学学报(医学版)

OBJECTIVE@#To determine the expressions of survivin and proliferating cell nuclear antigen (PCNA)in non-small cell lung cancer (NSCLC) and to explore its clinical pathological significance.@*METHODS@#Immunohistochemical SP method was used to detect the expressions of survivin and PCNA in 43 patients with NSCLC and 15 normal epithelial tissues of the lung. PCNA labeling proliferative index was assessed. Forty-three patients with NSCLC were followed up for more than 5 years.@*RESULTS@#The positive expression of survivin in NSCLC (79.1%) was significantly higher than that in normal epithelial tissues of the lung (P 0.05). The mean proliferative index of PCNA in NSCLC was much higher than that in normal epithelial tissues of the lung (P < 0.01). A positive correlation was present between the proliferative index and the tumor size, lymph node metastase, and clinical stage (P <0.01), while a negative correlation between the proliferative index and survival time (P <0.01). There was no correlation between proliferative index and age, sex, site, histological type and grade. The proliferative index was larger in patients with moderate or strong positive survivin expression than that in patients with negative or weak survivin expression (P < 0.05).@*CONCLUSION@#Over expression of survivin and PCNA is closely correlated to the progression and prognosis of patients with NSCLC, which is helpful to evaluate the progression of cancer and to predict the prognosis of NSCLC. The up-regulation of survivin expression and its close relationship with the cell proliferation in NSCLC suggest that survivin may play an important role in the carcinogenesis and development of lung cancer.

Proliferation inhibition of human lung adenocarcinoma cell line A549 transfected by RASSF1A gene / 中南大学学报(医学版)

OBJECTIVE@#To determine the effects of exogenous RASSF1A gene on the proliferation and expression of P65 and subunit of NF-kappaB, in lung adenocarcinoma cell line A549.@*METHODS@#pcDNA3.0-RASSF1A and pcDNA3. 0 were introduced into A549 cell line by lipofectin transfection, and the A549 cells stably expressing RASSF1A gene were established by G418 selection. The expression of RASSF1A was detected by Western blotting. The cytobiologic characterizations of the positive clone were analyzed by methythiazoletertraolium (MTT) assay and cytometry. The expressing of P65 was analyzed by RT-PCR and Western blotting.@*RESULTS@#A549 cells stably expressing RASSF1A protein were established by lipofection mediated transfection and selected for further study. Compared with the nontransfected and vector transfected cells, the positive clone cells grew more slowly. Flow cytometric data showed that more positive clone cells went into phase G0/G1 and fewer cells went into phase S. The expression of P65 in nuclear protein in positive clone cells was lower than that of the control group while there was no obvious difference between the expression of p65 mRNA and P65 protein in total protein among the 3 groups.@*CONCLUSION@#RASSF1A gene might suppress the proliferation of A549 cells through blocking the activity of P65 protein.