MMP-1/TIMP-1 expressions in rectal submucosa of females with obstructed defecation syndrome associated with internal rectal prolapse.

OBJECTIVE: To explore the MMP-1/TIMP-1 expressions in rectal submucosa of females with obstructed defecation syndrome (ODS) associated with internal rectal prolapse (IRP). METHODS: Fifty-six female patients with ODS associated with IRP were enrolled as Case group, and 43 female hemorrhoids of stages III-IV without constipation and IRP were enrolled as Control group. Quantitative reverse transcription polymerase chain reaction (qRT-PCR) and immunohistochemistry were performed to test the expressions of MMP-1/TIMP-1 in the rectal submucosa. Western blotting was used to examine protein expressions of MMP-1/TIMP-1 and pro-inflammatory cytokines (IL-6 and TNF-α) in the rectal submucosa. EVG staining was conducted to detect collagen and elastic fibers in rectal submucosa. RESULTS: The increased expression of MMP-1 was negatively linked to the decreased TIMP-1 level in the rectal submucosa of patients with ODS associated with IRP. Besides, the expressions of IL-6 and TNF-α were increased in the Case group as compared with the Control group. Additionally, ODS severity and the pro-inflammatory cytokines was positively linked to MMP-1, but negatively related to TIMP-1 in Case group. EVG staining showed that the area ratios of collagen and elastic fibers were lower in Case group than Control group. Through Pearson's correlation analysis, the area ratios of collagen and elastic fibers were positively associated with MMP-1 expression, but negatively correlated with TIMP-1 expression in rectal submucosa of patients with ODS associated with IRP. CONCLUSION: Elevated MMP-1 and reduced TIMP-1 were found in ODS associated with IRP, which was related to the ODS severity, inflammation and contents of collagen and elastic fibers.

Role of miR-22 in intestinal mucosa tissues and peripheral blood CD4+ T cells of inflammatory bowel disease.

OBJECTIVE: miR-22 is known to be involved in the pathogenesis of several autoimmune diseases, but it remains unclear whether miR-22 is associated with inflammatory intestinal disease (IBD). METHODS: The patients with ulcerative colitis (UC) and Crohn's disease (CD) were enrolled in this study. After the CD4+ T cells from healthy controls and active IBD patients were isolated and then transfected with miR-22 mimics/inhibitors, Quantitative real-time polymerase chain reaction (qRT-PCR) was conducted to measure expressions of miR-22, HDAC4, specific transcription factors in intestinal mucosa tissue and CD4+ T cells, while enzyme-linked immuno sorbent assay (ELISA) to detect expressions of inflammatory cytokines in PB. Antisense miR-22 was administered into mice during trinitrobenzene sulphoni cacid (TNBS)-induced colitis to determine its role in IBD. RESULTS: A significant elevation of miR-22 but an evident decrease of HDAC4 was found in CD4+ T cells in PB and intestinal mucosa tissues from IBD patients. In addition, there was a great reduction in HDAC4 and a dramatic enhancement in Th17 cell specific transcription factor (RORC) and inflammatory cytokines (IL-17A, IL-6 and TNF-α) after overexpression miR-22, which was opposite to the effect of inhibition of miR-22. Furthermore, administration of antisense miR-22 in TNBS-induced mouse colitis model significantly decreased numbers of interleukin (IL)-17A+ CD4+ T cells and the expressions of IL-17A, RORC, IL-6 and TNF-α. CONCLUSION: MiR-22 was up-regulated in CD4+ T cells in PB and intestinal mucosa tissues of IBD patients, which could promote Th17 cell differentiation via targeting HDAC4 to be involved in IBD progression.

SERPINA3 Silencing Inhibits the Migration, Invasion, and Liver Metastasis of Colon Cancer Cells.

OBJECTIVE: To investigate the impact of SERPINA3 on the migration, invasion, and liver metastasis of colon cancer cells. METHODS: Immunohistochemical staining was conducted to determine SERPINA3 expression in the cancer and adjacent normal tissues of 131 patients suffering from colon cancer. In vitro experiment, colon cancer cells with low (HT-29P), intermediate (KM-12C), and high (HT-29LMM, KM-12L4) metastatic potential were obtained to examine SERPINA3 expression levels. Besides, quantitative real-time PCR (qRT-PCR) and Western Blot were performed to detect SERPINA3 expression in HT-29LMM and KM-12L4 cells transfected with SERPINA3 siRNA; Wound-healing and Transwell assays to measure cell migration and invasion, respectively; and ELISA to detect MMP-2 and MMP-9 levels. In vivo experiment, mice with liver metastasis of colon cancer were established to observe the effect of SERPINA3 silencing on liver metastasis. Immunohistochemical assay was applied to evaluate the expressions of Serpina3, Mmp-2, Mmp-9, and proliferating cell nuclear antigen (Pcna) in liver metastasis tissues. RESULTS: SERPINA3 in colon cancer tissues was higher than in adjacent normal tissues, which was associated with patients' clinicopathological features. Besides, SERPINA3 expression showed a rising trend in low, intermediate, and high metastatic potential colon cancer cells. After KM-12L4 and HT-29LMM cells transfected with SERPINA3 siRNA, the migration and invasive ability of cells, as well as the expression levels of MMP-2 and MMP-9 were all decreased. Moreover, SERPINA3 siRNA could not only reduce live metastasis of mice, but also down-regulate the expression of Mmp-2 and Mmp-9 in liver metastasis tissues. CONCLUSION: SERPINA3 silencing could inhibit the migration, invasion, and liver metastasis of colon cancer cells.