Anti-TGFB1 monoclonal antibody, clone 3Bs3

Background: Cholangiocarcinoma (CCA) is characterized by early lymphatic, metastasis, and low survival rate. Epithelial-mesenchymal transition (EMT) is able to induce tumor metastasis. Although the TGF-beta/miR-200 signals promote EMT in various types of cancer, the regulatory mechanism in CCA is still unclear. Methods: Expression of miR-200b, TGF-beta, and EMT markers were measured in tumor samples and cell lines by qRT-PCR and western blot. CCK8 assay was performed to measure the cell viability. Transwell assay was used to evaluate migration and invasion. The target genes of miR-200b and transcription factor of TGF-beta were analyzed using dual-luciferase reporter system. Results: We have demonstrated that CCA exhibited remarkable EMT phenotype and miR-200b was reduced in CCA patients (n = 20) and negatively correlated to TGF-beta. Moreover, two CCA cells, HCCC, and RBE, with epithelial appearances treated with TGF-beta, showed fibroblastic-like cell morphology with downregulated miR-200b expression. Forced expression of miR-200b abrogated TGF-beta-induced EMT initiation, with decreased cell proliferation, migration, and invasion in vitro. Also, TFAP2A (encode AP-2 alpha) and MAPK7 were found to be targeted by miR-200b to downregulate EMT and AP-2 alpha inhibited miR-200b by directly promoting transcription of TGFB1. Overexpression of MAPK7 significantly reversed miR-200b-induced inhibition of EMT, migration, and proliferation by increasing the expression of TGF-beta, cyclin D1, and Cdk2. Further, the administration of miR-200b induced a remarkably tumor regression in vivo and reduced the effect of TGF-beta-related EMT in AP-2 alpha and MAPK7-dependent manner. Conclusions: Our study highlights that miR-200b-based gene therapy is effective in the treatment of CCA.

Scope Methods This study examines the beneficial effects of Goji berry against spontaneous colitis and its prebiotic role in IL-10-deficient mice. IL-10-deficient mice are assigned to a standard rodent diet (control) or a control diet supplemented with Goji (1% of dry feed weight) for 10 weeks, at which point colonic tissues and fecal contents are collected. Results Conclusion Goji supplementation decreases colonic pathobiological scores and mRNA expression of Il17a and Tgfb1, while it enhances Muc1 expression and fecal IgA content. Illumina MiSeq sequencing reveals that Goji supplementation increases Actinobacteria phylum, resulting in a bloom of Bifidobacteria in gut microbiota. Additionally, dietary Goji promotes butyrate-producing bacteria including Lachnospiraceae-Ruminococcaceae family and Roseburia spp. under Clostridium cluster XIVa. Furthermore, butyrate-producers Clostridium leptum and its dominant constituent Fecalibacterium prazusnitzii are markedly increased in the Goji group. Moreover, the gene-encoding butyryl-coenzyme A CoA transferase, a key enzyme responsible for butyrate synthesis in butyrate-producing bacteria, is increased sixfold in the fecal samples of Goji group associated with increased fecal butyrate content. Data collectively show that dietary Goji results in the blooming of Bifidobacteria and butyrate-producing bacteria. These bacteria may cross-feed each other, conferring preventative effects against colitis in IL-10-deficient mice.