ABSTRACT
OBJECTIVES@#Ulcerative colitis (UC) is a chronic, relapsing inflammation of the colon. Impaired epithelial repair is an important biological features of UC. Accelerating intestinal epithelial repair to achieve endoscopic mucosal healing has become a key goal in UC. Yes-associated protein (YAP) is a key transcriptional coactivator that regulates organ size, tissue growth and tumorigenesis. Growing studies have focused on the role of YAP in intestinal epithelial regeneration. This study explore the molecular mechanism for the role YAP in modulating colonic epithelial proliferation, repair, and the development of colitis associated cancer.@*METHODS@#We constructed the acute colitis mouse model through successive 5 days of 3% dextran sulfate sodium salt (DSS) induction. Then YAP-overexpressed mouse model was constructed by intraperitoneal injection the YAP overexpressed and negative control lentivirus into DSS mice. On the 5th day of DSS induction and the 5th day of normal drinking water after removing DSS (5+5 d), the mice were killed by spinal dislocation. The colon was taken to measure the length, and the bowel 1-2 cm near the anal canal was selected for immunohistochemical and Western blotting. We used YAP over-expressed colonic epithelial cells and small interfering signal transducer and activator of transcription 3 (STAT3) RNA to probe the regulation of YAP on STAT3, using cell counting kit-8 and scratch assays to explore the role of YAP on colonic epithelial cell proliferation. Finally, we conducted co-immunoprecipitation to test the relationship between YAP and STAT3.@*RESULTS@#After DSS treatment, the expression of YAP was dramatically diminished in crypts. Compared with the empty control mice, overexpression of YAP drastically accelerated epithelial regeneration after DSS induced colitis, presenting with more intact of structural integrity in intestinal epithelium and a reduction in the number of inflammatory cells in the mucosa. Further Western blotting, functional experiment and co-immunoprecipitation analyses showed that the expression of YAP in nucleus was significantly increased by 2 h post DSS cessation, accompanied with up-regulated total protein levels of STAT3 and phosphorylated-STAT3 (p-STAT3). Overexpression of YAP enhanced the expression of STAT3, p-STAT3, and their transcriptional targets including c-Myc and Cyclin D1. In addition, it promoted the proliferation and the "wound healing" of colonic cells. However, these effects were reversed when silencing STAT3 on YAP-overexpressed FHC cells. Moreover, protein immunoprecipitation indicated that YAP could directly interact with STAT3 in the nucleus, up-regulatvng the expressvon of STAT3. Finally, during the process of CAC, overexpression of YAP mutant caused the down-regulated expression of STAT3 and inhibited the development and progress of CAC.@*CONCLUSIONS@#YAP activates STAT3 signaling in regulation of epithelial cell proliferation and promotes mucosal regeneration after DSS induced colitis, which may serve as a potential therapeutic target in UC. However, persistent and excessive YAP activation may promote CAC development.
Subject(s)
Animals , Mice , Cell Proliferation , Colitis/drug therapy , Colon/metabolism , Dextran Sulfate/adverse effects , Disease Models, Animal , Intestinal Mucosa , Mice, Inbred C57BL , Neoplasm Recurrence, Local/metabolism , STAT3 Transcription Factor/metabolism , YAP-Signaling Proteins/metabolismABSTRACT
Objective To clarify the role of BRAFV600E and TERT promoter mutations in cervical lymph node metastasis in papillary thyroid carcinoma.Methods The data of 432 patients with thyroid papillary carcinoma who underwent surgery from February 2017 to September 2017 at the First Affiliated Hospital of Zhengzhou University were analyzed retrospectively.The mutation of BRAFV600E and TERT promoter was detected by Sanger sequencing.The effect of BRAFV600E and TERT on cervical lymph node metastasis in patients with papillary thyroid carcinoma was analyzed by Chi-square test.Results The mutation rates of BRAFV600E and TERT promoter were 77.8％ (336/ 432) and 5.3％ (23/432) respectively in 432 papillary thyroid carcinoma patients.The probability of cervical lymph node metastasis in patients with BRAFV600E mutation was significantly higher than that in non-mutation patients (P ＜ 0.05).The probability of cervical lymph node metastasis in patients with TERT promoter mutation was significantly higher than that in non-mutated patients (P ＜ 0.05).Patients with both BRAFV600E and TERT promotermutation had a significantly higher incidence of cervical lymph node metastases than patients with the BRAFV600E mutation alone (P ＜ 0.05).Conclusions The mutations of BRAFV600E and TERT promoter are closely relevant to the occurrence of cervical lymph node metastasis in papillary thyroid carcinoma.Preoperative fine-needle aspiration cytology and postoperative routine pathological molecular diagnosis can help clinicians to develop a more rational treatment strategy,and a more accurate assessment of the risk of relapse.
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Objective To investigate the diagnostic efficacy of fecal calprotectin(FC) on assessing endoscopic disease activity in colonic or ileo-colonic Crohn disease (CD) and CD-related surgery patients .Methods Totally 56 colonic or ileo-colonic CD pa-tients ,25 CD-related surgery patients and 25 irritable bowel syndrome (IBS) patients with previously confirmed diagnosis of CD and IBS were enrolled into this study .Fecal samples were collected from 1 to 3 day before bowel preparation and FC was measured by ELISA .Endoscopic activity was determined for colonic or ileo-colonic CD with Simple Endoscopic Score for Crohn′s Disease (SES-CD) and CD-related surgery patients with the Rutgeerts′ score .Results Among colonic or ileo-colonic CD patients ,the levels of FC in endoscopic active patients had significantly higher than that of endoscopic remission patients and IBS patients(P 0 .05) ,FC cutoff level of 250 μg /g gave a sensitivity and specificity of 50 .0% ,66 .7% ,respectively .Conclusion FC is a surrogate marker for the evaluation of colonic or ileo-colonic CD endoscopic disease activity .The FC ,however ,can not distinct remission period and active period after CD surgery .
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Objective To assess the performance of the methylation‐sensitive high‐resolution melting curve analysis (MS‐HRM analysis) on the detection of the methylation in stool DNA for colorectal cancer screening .Methods Eighty‐two qualified stool samples were collected from 27 patients with colorectal cancer patients (CRC group) ,25 patients with advanced adenomas (AA group) ,and 30 healthy people (control group) .The methylation status of vimentin gene in all of the stool samples was detected by the MS‐HRM analysis on the LightCycler 480 platform .The fecal occult blood test (FOBT) was also used for the same samples . Results The positive rates of the MS‐HRM assay in the CRC group ,AA group ,and control group were 81 .5% (22/27) ,80 .0%(20/25) ,and 6 .7% (2/30) respectively .The positive rates of FOBT in the three groups were 37 .0% (10/27) ,12 .0% (3/25) and 3 .3% (1/30) respectively .The diagnostic sensitivity of the MS‐HRM assay for colorectal cancer and advanced adenomas (80 .8% , 42/52) was significantly higher than that of FOBT (25 .0% ,13/52)(P0 .05) .Conclusion MS‐HRM performs better than FOBT and has great application potential in the detection of stool DNA methylation for colorectal cancer screening .
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Objective To study the effect of adeno associated virus hepatocyte growth factor K1(AAV‐HGFK1)on the prolifer‐ation of 4 different colorectal cell lines with or without KRAS or BRAF mutation .Methods The levels of epidermal growth factor receptor (EGFR) mRNA were determined in SW48 without KRAS or BRAF mutation ,Lovo with KRAS mutation ,SW620 with KRAS mutation ,HT29 with BRAF mutation by quantitative real time PCR ,respectively .After the infection of AAV‐HGFK1 ,the expressions of EGFR ,p‐EGFR and β‐actin were detected by Western blot and the proliferation of the cells were assayed using MTT .Results Lovo ,SW48 and HT29 expressed EGFR protein while SW620 did not .EGF promoted the proliferation of Lovo , SW48 and HT29 cells .AAV‐HGFK1 down‐regulated the phosphorylation of EGFR and significantly inhibited their proliferation . But EGF had no effect on proliferation of SW620 stimulated by EGF .Conclusion AAV‐HGFK1 exhibited its antitumor effects through EGF/EGFR signaling irrespective of the KRAS or BRAF mutation and may also act through other signaling pathways .