LncRNA H19 promotes epithelial mesenchymal transition and metastasis of esophageal cancer via STAT3/EZH2 axis.

BACKGROUND: Long non-coding RNA H19 (lncRNA H19) has been widely reported in esophageal cancer (EC), and previous study had found that lncRNAH19 was up-regulated in EC and promoted cell proliferation and metastasis. However, the mechanism still needs further studied. METHODS: Levels of lncRNA H19 were analyzed by qRT-PCR in matched samples from 30 patients. Expression levels of lncRNA H19, let-7, STAT3 and EZH2 were additionally identified by qRT-PCR and western blotting in five EC cell lines. The effects of lncRNA H19 on cell proliferation, migration, invasion and apoptosis in cell lines were performed by MTT assay, colony formation assay, Transwell assay and flow cytometry in vitro, and tumor formation was detected by xenograft nude mice model in vivo. The expression level of STAT3, EZH2, ß-catenin, and EMT and metastasis related molecules such as E-cadherin, N-cadherin, Snail-1 and MMP-9 was assessed by qRT-PCR and western blotting. Finally, luciferase reporter assay and RIP assay were used to verify the interaction between lncRNA H19 and let-7c, and their subsequent regulation of STAT3. RESULTS: Knockdown of lncRNA H19 repressed cell proliferation, migration and invasion as well as EMT and metastasis via STAT3-EZH2-ß-catenin pathway, while lncRNA H19 regulated STAT3 negatively regulated let-7c in EC cell lines. CONCLUSIONS: lncRNA H19 facilitates EMT and metastasis of EC through let-7c/STAT3/EZH2/ß-catenin axis.

Increased connective tissue growth factor expression in a rat model of chronic heart allograft rejection.

BACKGROUND/PURPOSE: Chronic rejection limits the long-term success of cardiac transplantation and the underlying cause of the disease is unknown. Connective tissue growth factor (CTGF) is considered as a mitogenic and chemotactic factor for fibroblasts, and is associated with cell proliferation and collagen synthesis. We evaluated the expression of CTGF in a rat model of heart allograft chronic rejection. METHODS: Intra-abdominal heterotopic heart transplantation was performed from 20 Wistar rats to 20 Sprague-Dawley (SD) rats that received cyclosporine, mycophenolate mofetil and methylprednisolone as immunosuppression. Ten heart allografts were explanted at 2 and 8 weeks postoperatively for analysis of morphologic changes. The hearts from 10 normal Wistar rats served as a control group. Coronary artery density, luminal loss of myocardial coronary arteries, and myocardial fibrosis were measured. The expression of CTGF was studied by immunohistochemistry. Correlation between CTGF expression and development of cardiac allograft vasculopathy (CAV) or fibrosis was studied. RESULTS: Allografts harvested at 8 weeks postoperatively showed more coronary intimal proliferation, fibrosis and CTGF expression compared with the 2-week allografts (p < 0.05) and the controls (p < 0.01), but the coronary artery density was lower than in the control group (p < 0.05). However, the control group showed negligible CTGF expression. There were strong negative correlations between the gray value of CTGF protein expression and cardiac fibrosis and coronary intimal occlusion (r = -0.734, -0.713; p < 0.01), which demonstrated that CTGF protein expression was positively correlated with cardiac fibrosis and coronary intimal occlusion. CONCLUSION: CTGF is expressed in cardiomyocytes in CAV. Increased expression of CTGF in cardiac allografts is associated with development of CAV and fibrosis formation.

Expression of connective tissue growth factor in acute heart allograft rejection in rats.

OBJECTIVE: To detect the expression of connective tissue growth factor (CTGF) in acute heart allograft rejection in rats and to investigate the relationship between CTGF expression and cardiac allograft fibrosis. METHODS: Sixteen Wister rats served as donors and another 16 Sprague-Dawely (SD) rats served as recipients. Intra-abdominal heterotopic heart transplantation was performed. All rats received 10 mg/(kg.d) cyclosporine, 40 mg/(kg.d) CellCept, and 3 mg/(kg.d)methylprednisolone immunosuppression after the surgery. Ten allografts were harvested 2 weeks postoperation while 10 normal Wister rats served as controls. The paraffin sections of harvested heart specimens were stained with hematoxylin and eosin (HE),and van Gieson(VG) for the examination of morphological changes to observe the lumen loss of myocardial coronary arteries and myocardial fibrosis. The expression of CTGF was studied by immunnohistochemical method and was measured semi quantitatively. The correlation between the CTGF expression and allograft fibrosis was studied. RESULTS: The allografts showed a typical symbol of acute rejection with excessive granulocyte infiltration around the vessel wall and myocardial interstice. There were also intimal proliferation and obvious fibrosis in the acute group and the differences between the acute and control group were significant (P< 0.05). The expression of CTGF protein was mainly located around the vascular and myocardial lesions in the acute group while the control group showed no CTGF expression. The gray scale value of CTGF was (AR vs NH: 103.52+/-6.42 vs. 182.61+/-8.72, P< 0.05). Strong negative correlations were found between the gray scale value and fibrosis formation(r=-0.734, P< 0.01). CONCLUSION: CTGF was overexpressed in acute allograft rejection rat hearts and might be involved in the pathogenesis of transplanted heart fibrosis.

[Integrated mediastinal pleura upon aortic arch and left cervical anastomosis in radical operation for esophageal carcinoma].

OBJECTIVE: To evaluate the therapeutic effect of remaining integrated mediastinal pleura upon the aortic arch and performing the anastomosis at the left cervix in radical operation for esophageal carcinoma. METHODS: Ninety-eight patients with esophageal carcinoma were treated with the operation mentioned above. Among them, 56 patients had cancer in the middle, 12 in the upper-middle, 24 in the lower-middle segments, and 6 had double-primary tumors, with carcinoma length of (5.2+/- 2.4) cm. The TNM stages were 6 of Stage I and 92 of Stages II-III. All cases were squamous cell carcinomas. RESULTS: All patients had satisfactory operation processes, without perioperative death, chylothorax, dyspnea, gastric retention, incision infection, and severe gastro-esophageal reflux. The life quality of the patients was improved. CONCLUSION: The radical operation with remaining integrated mediastimal pleura upon the aortic arch and anastomosis at the cervix for treating esophageal carcinomas is of minimal invasion with fewer complications, and may be used in clinical practice.

[Connective tissue growth factor and pulmonary allograft fibrosis in rats].

OBJECTIVE: To investigate the role of connective tissue growth factor (CTGF) in pulmonary allograft fibrosis in rats. METHODS: The lungs of 20 Wistar rats were transplanted into 20 Sprague-Dawley(SD) rats. Ten allograft lungs were harvested 1 week postoperatively (acute rejection group,AR); the other 10 allografts were harvested 6 weeks postoperatively (chronic rejection group,CR); and ten normal Wistar rats served as a control group(normal lung, NL). Paraffin embedded sections of the harvested lung specimens were stained with hematoxylin and eosin (HE), Van Gieson (VG) for the examination of tissue morphology under the microscope. The scores of lung fibrosis were measured and the wet/dry ratio of the lung specimens was evaluated. The CTGF expression was determined by immunohistochemical method. RESULTS: The wet/dry ratios of lung decreased gradually(AR group vs. control group: 3.48+/-0.47 vs. 4.67+/-0.51, P<0.05; CR group vs. AR group: 2.85+/-0.52 vs. 3.48+/-0.47, P<0.05). The transplanted lungs showed massive lymphocytic infiltration, interstitial fibrosis, destroyed alveolus architecture, obliterative bronchiolitis, and lung tissue consolidation. These pathological changes were more severe in the CR group than in the AR group, but there were no such changes in the control group (scores of pulmonary fibrosis: NH, 0.00+/-0.00; AR, 0.98+/-0.47; CR, 2.35+/-0.52; AR vs. NH, P<0.01; CR vs. AR, P<0.01). CTGF was not expressed in the normal rat lungs (0.00+/-0.00); however, it was detected in the lung allograft after the operation. The CTGF expression in the CR group was significantly higher than that in the AR group (P<0.01). CONCLUSION: The expression of CTGF protein is related to the transplanted pulmonary fibrosis,and is involved in the pathogenesis of transplanted pulmonary fibrosis.

[Expression of PTEN, p16, p21, and p53 proteins in lung cancer using tissue microarray and their clinical significance].

BACKGROUND & OBJECTIVE: The genesis of lung cancer was associated with mutation or abnormal expression of PTEN, p16, p21, and p53. Tissue microarray provides a high throughout tool for genes expression. But little is reported about expression of PTEN, p16, p21, and p53 in lung cancers with tissue microarray. The aim of this study was to investigate the expression of PTEN, p16, p21, and p53 proteins and to analyze their relationship with the pathogenesis, invasion, and metastasis in lung cancer. METHODS: The expression of the antioncogene proteins in 100 cases of lung cancer and corresponding adjacent tissues were determined by tissue microarray combined with immunohistochemistry. RESULTS: The positive expression rates of PTEN, p16, p21, and p53 proteins were 31% (31/100), 38% (38/100), 42% (42/100), 53% (53/100) in lung cancer tissues, and were 85% (85/100), 72% (72/100), 80% (80/100), and 23% (23/100) in the adjacent cancer tissues, respectively, showing a low expression of PTEN, p16, p21 in cancer tissues, and high expression of p53 outside of them (P< 0.05, P< 0.01). Furthermore, the expression of PTEN, P16, and p53 proteins showed positive correlation with the clinical degrees and pathological stages of lung squamous carcinomas and adenocarcinomas (P< 0.05,P< 0.01). In lung cancer with lymph node metastasis, the expression of PTEN, p16, and p21 were low, but the expression of p53 increased significantly (P< 0.05, P< 0.01). CONCLUSION: Tissue microarray provided a useful high-throughout tool for multigene expression in large-scale investigations. There existed low expression of PTEN, p16, p21 proteins and over-expression of mutated p53 protein. Coexpression of these antioncogenes played an important role in invasion and metastasis in lung cancer.