4792

NFKBIA

OBJECTIVE: To investigate the expression of inhibitor of nuclear factor kappaBalpha (IkappaBalpha) in bladder cancer cell lines and its correlation with epithelial-mesenchymal transition (EMT) and invasive potential in vitro. METHODS: The expressions of IkappaBalpha, epithelia cadherin (E-cadherin), neuronal cadherin (N-cadherin) and vimentin in human bladder cancer RT4, 5637, 235J, J82 and T24 cell lines were detected by Western blotting. Invasive potential of these cell lines was evaluated by Transwell(TM) assay. In addition, morphological characteristics of those cells were carefully observed under a microscope. RESULTS: RT4 and 5637 cells exhibited epithelial cell morphological features, but 253J, J82 and T24 cells had mesenchymal cell morphological features. E-cadherin was over-expressed in RT4 and 5637 cell lines, but the expressions of vimentin and N-cadherin could not been detected in the two cells. E-cadherin could be detected in 253J cells with a lower expression level; J82 and T24 cells had no expression of E-cadherin; vimentin and N-cadherin were highly expressed in 253J, J82 and T24 cells. Transwell(TM) assay showed higher invasive potential of 253J, J82 and T24 cells than RT4 and 5637 cells in vitro. In addition, RT4 and 5637 cells expressed higher levels of IkappaBalpha than 253J, J82 and T24 cells. CONCLUSION: expression of IkappaBalpha in human bladder cancer cells is negatively correlated with EMT and tumor invasion in vitro.

BACKGROUND: MicroRNA-196a-5p (miR-196a-5p) has been reported to be involved in the metastatic process of several cancers. In present work, we aimed to investigate the effects of miR-196a-5p and its potential target IkappaBalpha on migration, invasion and epithelial-mesenchymal transition (EMT) of colorectal cancer (CRC) cells. METHODS: CCK-8 assay, wound healing assay and cell invasion assay were performed to evaluate the cell proliferation, migration and invasion. In vivo metastasis models were used to investigate the tumor metastasis ability. Real-time PCR, immunofluorescence staining or western blot were utilized to detect the expression of miR-196a-5p, IkappaBalpha, p-IkappaBalpha, nuclear p65 and EMT markers including E-cadherin, N-cadherin and fibronectin. Dual luciferase reporter assay was carried out to determine whether there is a direct interaction between miR-196a-5p and IkappaBalpha mRNA. RESULTS: Using SW480 cell with miR-196-5p over-expressed plus SW620 and HCT116 cells with miR-196a-5p knockdown, we found that miR-196a-5p promoted cell proliferation, migration and invasion in vitro and facilitated liver metastasis in vivo. We also observed that miR-196a-5p knockdown or NF-kappaB pathway inhibition up-regulated E-cadherin while down-regulated N-cadherin and fibronectin. By contrast, miR-196a-5p over-expression promoted EMT process of CRC. Data of dual luciferase reporter assay indicated that miR-196a-5p targeted the IkappaBalpha. Moreover, miR-196a-5p down-regulated IkappaBalpha expression while up-regulated nuclear p65 expression. Additionally, over-expression of IkappaBalpha in CRC cells attenuated the effects of miR-196a-5p on cell migration, invasion and EMT. CONCLUSIONS: miR-196a-5p may play a key role in EMT, invasion and metastasis of CRC cells via targeting the IkappaBalpha.