MiR 362 upregulation promoted cell proliferation and induced apoptosis resistance in gastric cancer To investigate the biological impact of miR 362 upregula tion on gastric cancer progression, the BGC 823 and SGC 7901 gastric cancer cell lines had been used to stably ex press miR 362. MTT assay showed that miR 362 upregu lation substantially enhanced the price of cell proliferation, and this was confirmed by colony formation assay. Flow cytometry revealed a dramatic increase in the percentage of S phase cells in miR 362 overexpressing BGC 823 and SGC 7901 cells as compared with manage BGC 823 and SGC 7901 cells, respectively. Annexin V and TUNEL staining demonstrated that miR 362 overexpression augmented the resistance of gastric cancer cells to apoptosis induced by the cisplatin remedy.
These outcomes suggest that miR 362 plays an oncogenic role in gastric cancer cells in vitro. MiR 362 inhibition decreased cell proliferation and induced apoptosis in human gastric cancer We examined the impact of miR 362 inhibition on gastric cancer progression. Constant using the above results, the MTT and colony formation assays showed kinase inhibitor Nutlin-3b that miR 362 suppression significantly inhibited the development price of both BGC 823 and SGC 7901 cells as compared with that of control cells. Flow cytometry showed that miR 362 inhibition decreased the percentage of cells in S phase peak but enhanced that of G1 G0 phase cells, suggesting that miR 362 inhibition outcomes in G1 S arrest in gastric cancer cells. Annexin V and TUNEL staining demonstrated that miR 362 inhibition decreased resistance to apoptosis in cisplatin treated gastric cancer cells.
MiR 362 activated the NF B pathway We investigated the underlying molecular mechanism that may well be accountable for the oncogenic roles of miR 362. As the NF B signaling pathway is regularly found hyperactivated in gastric tumors, and activation of NF B signaling induces cell proliferation and apoptosis resistance, we investigated whether or not miR 362 regulated NF B activity. NF B selleck chemical reporter lucifer ase activity and the expression levels from the eight NF B target genes have been drastically enhanced in miR 362 more than expressing cells, but have been decreased in cells in which miR 362 had been inhibited.
Even though miR 362 had no effect on the total NF B p65 protein expression, cellular fractionation and immunofluores cence staining showed that miR 362 overexpression promoted nuclear accumulation of NF B p65, whilst miR 362 inhibition reduced nuclear NF B p65 expres sion, indicating that miR 362 activates the NF B pathway by means of promotion of nuclear NF B accumulation. Inhibition of NF B signaling by the trans fection of an IB dominant adverse mutant led to a dramatic lower in S phase peak cells but elevated the G0 G1 phase peak population and cisplatin sensitivity in miR 362 overexpressing cells, suggesting that NF B pathway activation is function ally relevant to miR 362 mediated proliferation and anti apoptosis.