[LKB1 regulates epithelial-mesenchymal transition in Peutz-Jeghers hamartoma and intestinal epithelial cells].

OBJECTIVE: To investigate the molecular mechanism by which LKB1 regulates epithelial-mesenchymal transition (EMT) in Peutz-Jeghers hamartoma and intestinal epithelial cells. METHODS: Immunohistochemistry was used to detect gene expression of LKB1, E-cadherin, and vimentin in 20 hamartoma tissues and 10 normal intestinal tissues, and collagen fiber deposition was analyzed using Masson trichrome staining. Normal intestinal epithelial NCM460 cells were transfected with LKB1 shRNA plasmid or negative control via lentiviral vectors, and the role of LKB1 in cell polarization and migration were determined using CCK8 and Transwell assays. Western blotting, quantitative real-time PCR (qPCR) and immunofluorescence were used to assess the alterations of EMT markers in the cells with LKB1 knockdown. RESULTS: Compared with normal intestinal tissues, hamartoma polyps showed significantly decreased LKB1 and E-cadherin expressions and increased vimentin expression with increased collagen fiber deposition. The cells with LKB1 knockdown exhibited enhanced cell proliferation and migration activities (P<0.01). Western blot analysis, qPCR and immunofluorescence all detected decreased E-cadherin and increased N-cadherin, vimentin, Snail, and Slug expressions in the cells with LKB1 knockdown. CONCLUSION: s LKB1 deficiency triggers EMT in intestinal epithelial cells and Peutz-Jeghers hamartoma, suggesting that EMT can serve as the therapeutic target for treatment of Peutz-Jeghers syndrome.

[Sample targeting under stereomicroscopy can raise the detection rate of early colorectal cancer].

OBJECTIVE: To establish a set of pathological diagnosis method to raise the detection rate of early colorectal cancer. METHODS: All patients with colorectal tumor underwent ordinary electron enteroscopy in 2005. The lesions larger than 10 mm underwent indigo carmine staining and magnifying electron enteroscopy to observe the pit pattern. Endoscopic mucosal resection (EMR) or endoscopic piecemeal resection (EPMR) was performed on the suspected cases of cancer, such as laterally spreading tumor (LST). The resected specimens were stained with cresyl violet and observed by stereomicroscopy to determine the pit patterns. The parts showing the pit patterns associated with early colorectal cancer were targeted and biopsy specimens collected here to undergo pathohistological examination. Routine pathological examination was conducted on the other parts of the same specimen as control. The results of these specimens were compared with those of the specimens collected by ordinary methods from the patients with colorectal tumor in 2004. RESULTS: In 2005 40 patients with colorectal tumor were suspected as with cancer and underwent EMR or EPMR of which 16 were confirmed to be with early stage colorectal cancer, including severe dysplasia by sampling targeting (40%). And the routine pathohistological examination of the randomly collected parts from these same specimens showed 15 cases of mild or moderate dysplasia and only one case of severe dysplasia, with a detection rate of 2.5%, significantly lower than that of the result of sample targeting under stereomicroscopy (P < 0.01). In 2004, out of the 54 patients suspected to be with colorectal cancer only 4 cases of early cancer, including severe dysplasia were detected with a detection rate of 7.4%, significantly than that of the year 2005 (P < 0.01). CONCLUSION: Sample targeting and localized biopsy under stereomicroscopy raises the detection rate of early colorectal cancer.

Training in early gastric cancer diagnosis improves the detection rate of early gastric cancer: an observational study in China.

Few studies have analyzed the training of endoscopists in the diagnosis of early gastric cancer (EGC). This study assessed whether specific training of endoscopists improves the detection rate of EGC. The rates of detection of EGC by endoscopists at the Digestive Endoscopy Center of the Affiliated Nanfang Hospital of China Southern Medical University between January 2013 and May 2014 were retrospectively analyzed. Because some endoscopists received training in the diagnosis of EGC, beginning in September 2013, the study was divided into 3 time periods: January to September 2013 (period 1), September 2013 to January 2014 (period 2), and January to May 2014 (period 3). The rates of EGC detection during these 3 periods were analyzed. From January 2013 to May 2014, a total of 25,314 gastroscopy examinations were performed at our center, with 48 of these examinations (0.2%) detecting EGCs, accounting for 12.1% (48/396) of the total number of gastric cancers detected. The EGC detection rates by trained endoscopists during periods 1, 2, and 3 were 0.3%, 0.6%, and 1.5%, respectively, accounting for 22.0%, 39.0%, and 60.0%, respectively, of the gastric cancers detected during these time periods. In comparison, the EGC detection rates by untrained endoscopists during periods 1, 2, and 3 were 0.05%, 0.08%, and 0.10%, respectively, accounting for 3.1%, 6.0%, and 5.7%, respectively, of the gastric cancers detected during these times. After training, the detection rate by some trained endoscopists markedly increased from 0.2% during period 1 to 2.3% during period 3. Further, the use of magnifying endoscopy with narrow-band imaging (M-NBI) (odds ratio = 3.1, 95% confidence interval 2.4-4.1, P < 0.001) contributed to the diagnosis of EGC. In conclusion, specific training could improve the endoscopic detection rate of EGC. M-NBI contributed to the diagnosis of EGC.

Low molecular weight heparin relieves experimental colitis in mice by downregulating IL-1ß and inhibiting syndecan-1 shedding in the intestinal mucosa.

Low molecular weight heparin (LMWH) exhibits anti-inflammatory properties, but its effect on inflammation in colitis remains unclear. This study aimed to evaluate the therapeutic effects of LMWH on dextran sulfate sodium (DSS)-induced colitis in mice, in which acute colitis progresses to chronic colitis, and to explore the potential mechanism involved in this process. C57BL/6 mice were randomly divided into control, DSS, and DSS plus LMWH groups (n = 18). Disease activity was scored by a disease activity index (DAI). Histological changes were evaluated by hematoxylin and eosin (HE) staining. The mRNA levels of syndecan-1, interleukin (IL)-1ß, and IL-10 were determined by quantitative reverse transcription polymerase chain reaction. Protein expression of syndecan-1 was detected by immunohistochemistry. The serum syndecan-1 level was examined by a dot immunobinding assay. LMWH ameliorated the disease activity of colitis induced by DSS administration in mice. Colon destruction with the appearance of crypt damage, goblet cell loss, and a larger ulcer was found on day 12 after DSS administration, which was greatly relieved by the treatment of LMWH. LMWH upregulated syndecan-1 expression in the intestinal mucosa and reduced the serum syndecan-1 level on days 12 and 20 after DSS administration (P<0.05 vs. DSS group). In addition, LMWH significantly decreased the expression of both IL-1ß and IL-10 mRNA on days 12 and 20 (P<0.05 vs. DSS group). LMWH has therapeutic effects on colitis by downregulating inflammatory cytokines and inhibiting syndecan-1 shedding in the intestinal mucosa.

[Changes in the biological behaviors of colon cancer cells under simulated hypoxia in vitro].

OBJECTIVE: To investigate the reactivity of colon cancer cell line SW480 and CD133(+) SW480 subsets to hypoxia in vitro and the changes in the expressions of anti-apoptosis and angiogenesis genes. METHODS: SW480 cells was subjected to CoCl(2) exposure at varying concentrations and for different time lengths to induce hypoxia, and the protein expression of hypoxia induced factor 1α (HIF-1α) was detected by Western blotting. The CD133(+) SW480 cells were sorted by magnetic activated cell sorting (MACS) and their proportion was assayed by flow cytometry (FCM). The CD133(+) SW480 subsets were exposed to CoCl(2) at the optimal concentration with exposure time selected in terms of HIF-1α level, and their tumor stem cell sphere formation ability was evaluated. Real-time PCR was used to compare the mRNA expression levels of the surface markers of colon cancer stem cells (CD133 and PROM1), survivin, and vascular endothelial growth factor (VEGF). RESULTS: Exposure to 200 µmol/L CoCl(2) for 8 h resulted in the highest HIF-1α expression in SW480 cells, but the same exposure failed to induce HIF-1α expression in CD133(+) SW480 subsets. The CD133(+) SW480 subsets, after CoCl(2)-induced hypoxia, showed significantly enhanced ability of cell sphere formation. Hypoxia of SW480 cells caused significant increases in CD133, survivin and VEGF mRNA levels by 1.607∓0.103, 2.745∓0.370 and 3.798∓0.091 folds, respectively (P<0.05). CONCLUSION: CoCl(2) can simulate hypoxia in colon cancer cells in vitro to induce stable HIF-1α expression, which is concentration- and time-dependent. The hypoxia-stimulated tumor stem sells show an enhanced sphere formation and anti-apoptotic and anti-angiogenic abilities.