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Archive > Volume 18 Issue 5 > 2011,18(5):502-507. DOI:10.3872/j.issn.1007-385X.2011.5.007 Prev Next

Basic Research

P65 gene on invasion of human triplenegative breast cancer cell line MDAMB231

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    Abstract:
    Objective:To investigate the effect of targeting silence of P65 gene by miRNA on adhesion and invasion of human triplenegative breast cancer (TNBC) cell line MDAMB231 in vitro and its possible mechanism. Methods: Three pairs of miRNAs specifically targeting P65 gene (P65miRNA1, P65miRNA2 and P65miRNA3) were designed and transfected into MDAMB231 cells, the level of P65 protein was detected by Western blotting, and P65miRNA with best silencing result was selected and used in the following experiments. The adhesive and invasive abilities of MDAMB231 cells before and after transfection were measured by adhesion assay and Transwell assay, respectively. mRNA expression of matrix metalloproteinase2 (MMP2) and MMP9 were detected by RTPCR and the activities of MMP2 and MMP9 were examined by gelatin zymography assay. Results: P65miRNA1, P65miRNA2 and P65miRNA3 plasmids were successfully constructed, P65miRNA1 and P65miRNA2, but not P65miRNA3, tranfection strongly inhibited the expression of P65 protein in MDAMB231 cells (>80%). The adhesion of MDAMB231 cells was not significantly influenced by P65miRNA1 and P65miRNA2 transfection (P>0.05), while the invasive ability of MDAMB231 cells was significantly reduced after P65miRNA1 or P65miRNA2 transfection (0.371±0.039, 0.309±0.046 vs 0.698±0065, P<0.05). Besides, mRNA expression of MMP2 (0.281±0.018, 0.478±0.023 vs 1.056±0.072, 1.128±0.059, P<0.05) and MMP9 (0.193±0.013, 0.371±0.035 vs 1.206±0.069, 1.089±0.057, P<0.05) and activities of MMP2 and MMP9 in MDAMB231 cells after P65miRNA1 or P65miRNA2 transfection were significantly decreased. Conclusion: Targeting silence of P65 expression can inhibit in vitro invasion of TNBC MDAMB231cells, which is related to the downregulation of MMP2 and MMP9 expression and their activities.

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    Project Supported:
    Project supported by the Science and Technology Development Foundation of Hebei Province (No. 10276167)

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