Knockdown of Grp78 Gene Expression By Over Expression of Mir-181a in Human Bladder Cancer Cell Line; 5637

Background and aim: invasion and metastasis of the bladder tumor to other organs significantly reduce the survival of patients. investigating the molecular and cellular mechanisms of cancer that are responsible for cellular invasion is one of the most promising ways to invent new therapies. some intracellular molecules, such as grp78, mmps, microrna, which play key roles in promoting tumor growth, metastasis, invasion and tissue degradation have been identified as molecular targets. previous researches have indicated that mir-181a down regulates grp78 mrna by targeting its 3′utr. the aim of this study was to investigate the effect of mir-181a construct on targeting grp78 and consequently studying its effects on growth, migration and metastasis in 5637 cells line.methods: the sequence of microrna was designed and then synthesized and cloned into the aav vector. then, successful clones were transferred to the 5637 cell line by lipofectamin 2000, expression of grp78, mmp-2, mmp-9, timp-1, timp-2 genes and 5637 cell’s proliferation and migration rate were measured by rt-qpcr, mtt and scratch assay methods respectively. moreover gelatin zymography was performed to investigate mmp-2 and mmp-9 gelatinolytic activities.results: data showed that mir-181a, grp78 and mmp-2 transcripts increased and expression of mmp-9, timp-1 and timp-2 genes decreased, when compared to controls. the results of wound scratch assay and its analysis showed that mir-181a significantly reduced migration of 5636 bladder cancer cells. there was no toxicity associated with mir-181a in cultures. gelatin zymography analysis revealed that transfection of mir-181a caused meaningful increase in the activities of mmp-2 and mmp-9.conclusion: our data clearly demonstrated that the mir-181a reduced invasiveness and migration of 5637 bladder cancer cells regardless of increased activities of mmp-9 and mmp-2.