| Background and ObjectsUlcerative colitis (UC) is a form of inflammatory bowel disease (IBD) that causes inflammation and ulcers in the colon. The disease is a type of colitis, which is a group of diseases that cause inflammation of the colon, the largest section of the large intestine, either in segments or completely. The main symptom of active disease is diarrhea mixed with blood. Because of its complex clinical diagnosis and treatment, it was recognized as one of disease difficult to cure by the world health organization. It is important to investigate the molecular mechanism, and looking for an effective targeted clinical therapy.A few studies shows that autophagy is involved in UC or others IBD. Prvious study demonstrated that HSF2 is highly expressed in UC tissue. These suggest that HSF2 gene may play a important role in the pathogenesis of UC, and has the potential in molecular therapy.Materials and methods1, Protein level of HSF2 in 50 UC tissues and normal tissues were detected by immunohistochemical method, and then analyzed the relationship with clinical pathological characteristics. Additionally, Transmission electron microscopy (TEM) were used to detect autophagy, and then autophagy markers like LC3Ⅱ, beclinl and p62 were detected by RT-qPCR and Western blot.2, HT29 cell line were treated with different concentrations of sodium butyrate. HSF2 shRNA and over-expressed lentivirus plasmid were constructed and then infected cells. HSF2 expression levels were detected by RT-qPCR and Western blot. Autophagy levels were assessed by TEM, GFP-LC3 transfection, and levels of autophagy markers using RT-qPCR and Western blot.3, HT29 cell were induced autophagy by Sodium butyrate,The key protein of endoplasmic reticulum stress and insulin/IGF 1 signaling pathways were detected by western blot.4, HSF2 shRNA expression plasmid were constructed, then infected HT29 cell lines, which were treated with 5nM sodium butyrate.Proliferation were detemined by MTT; cell invasion ability was evaluated by transwell cell chamber experiment; And cells apoptosis were determined by annexin V-FITC/PI stain; And autophagy levels were assessed by TEM and Western blot.Results1, The positive ratio between UC (62%) and normal tissues (32%) have significant sense with P value<0.05. more bubble of autophagy and autophagosome were observed by TEM. The results from quantitative PCR and western blot showed that autophagy marker LC3II and beclin 1 expression is higher than that in normal group, and the expression of the substrate LC3——p62 significantly decreased.2, the levels of HSF2 and LC3Ⅱ increased following enhanced concentrations of sodium butyrate. Over-expressing HSF2 could reversed the increase of autophagsome, dots of GFP-LC3 and cheanges of LC3Ⅱ, beclinl and p62 leves, which all were induced by sodium butyrate.3, We also found HSF2 regulating authphagy in UC may be involved in activation of endoplasmic reticulum stress by BIP and GRP94 pathway, as well as insulin/IGF 1/mTOR signaling pathways.4, In vitro, sodium butyrate treatment could promote cell apoptosis, inhibit the proliferation and invasion of HT29, and overpression of HSF2 could antagonised the effect of sodium butyrate, and enhanced the leves of autophagy induced by sodium butyrate.Conclusions1,HSF2, which is highly expressed, accompanied by autophagy overpression, may be play an important role in UC patients,.2, HSF2, which could regulated autophagy induced by sodium butyrate in UC, play an important role, and the molecular mechanism may be involved in ERS (BIP and GRP94) and insulin/IGF1-mTORc signal network.3, HSF2 may be a protective factor for epithelial cell apoptosis and autophagy. |
PDF Full Text Request