gant-61 and Stomach-Neoplasms

To explore how GLI-1 affects the EMT induced by TGF-β1 in gastric cancer.. Following 24 hours of culture of SGC-7901 cells in presence of TGF-β1, we observed the changes in morphology as well as mRNA and protein expressions of GLI-1, E-cadherin and Vimentin by RT-PCR and Western blot. Transwell assay was conducted to evaluate the changes in invasion ability of SGC-7901 cells. Then, SGC-7901 cells were co-treated with TGF-β1 and GANT 61, and changes of the above indexes were also detected using the corresponding methods.. In presence of TGF-β1, EMT was initiated in SGC-7901 cells EMT with increased cell invasion ability, and the mRNA and protein expressions of E-cadherin were downregulated, while those of the GLI-1 and Vimentin were upregulated. Conversely, the co-treatment of TGF-β1 and GANT 61 suppressed the increased cell invasion ability induced only by TGF-β1, and the changes in mRNA and protein expressions of these factors were abolished.. We found that GLI-1 facilitates the EMT induced by TGF-β1 in SGC-7901 cells, which may serve as a potential target in developing the clinical treatment of gastric cancer.

Topics: Antigens, CD; Cadherins; Cell Line, Tumor; Cell Movement; Epithelial-Mesenchymal Transition; Gene Expression Regulation, Neoplastic; Humans; Neoplasm Invasiveness; Pyridines; Pyrimidines; Signal Transduction; Stomach Neoplasms; Transforming Growth Factor beta1; Vimentin; Zinc Finger Protein GLI1

Mesenchymal stem/stromal cells (MSCs) are known to home to sites of tumor microenvironments where they participate in the formation of the tumor microenvironment and to interplay with tumor cells. However, the potential functional effects of MSCs on tumor cell growth are controversial. Here, we, from the view of bone marrow MSC-derived exosomes, study the molecular mechanism of MSCs on the growth of human osteosarcoma and human gastric cancer cells.. MSCs derived from human bone marrow (hBMSCs) were isolated and cultured in complete DMEM/F12 supplemented with 10% exosome-depleted fetal bovine serum and 1% penicillin-streptomycin, cell culture supernatants containing exosomes were harvested and exosome purification was performed by ultracentrifugation. Osteosarcoma (MG63) and gastric cancer (SGC7901) cells, respectively, were treated with hBMSC-derived exosomes in the presence or absence of a small molecule inhibitor of Hedgehog pathway. Cell viability was measured by transwell invasion assay, scratch migration assay and CCK-8 test. The expression of the signaling molecules Smoothened, Patched-1, Gli1 and the ligand Shh were tested by western blot and RT-PCR.. In this study, we found that hBMSC-derived exosomes promoted MG63 and SGC7901 cell growth through the activation of Hedgehog signaling pathway. Inhibition of Hedgehog signaling pathway significantly suppressed the process of hBMSC-derived exosomes on tumor growth.. Our findings demonstrated the new roles of hedgehog signaling pathway in the hBMSCs-derived exosomes induced tumor progression.

Topics: Blotting, Western; Bone Marrow Cells; Cell Movement; Cell Survival; Cells, Cultured; Exosomes; Hedgehog Proteins; Humans; Mesenchymal Stem Cells; Microscopy, Confocal; Microscopy, Electron, Transmission; Osteosarcoma; Patched-1 Receptor; Pyridines; Pyrimidines; Real-Time Polymerase Chain Reaction; Signal Transduction; Smoothened Receptor; Stomach Neoplasms; Tumor Microenvironment; Zinc Finger Protein GLI1