中国肿瘤生物治疗杂志2011，Vol.18Issue(5)：502-507,6.DOI:10.3872/j.issn.1007-385X.2011.05.007
P65基因对人三阴性乳腺癌细胞MDA-MB-231体外侵袭的影响
Effect of P65 gene on invasion of human triple-negative breast cancer cell line MDA-MB-231
摘要
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-l, 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-l, P65miRNA-2 and P65miRNA-3 plasmids were successfully constructed, P65miRNA-l 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-l and P65miRNA-2 transfection (P>0.05), while the invasive ability of MDA-MB-231 cells was significantly reduced after P65miRNA-l 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 t? 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 ofMMP-2 and MMP-9 in MDA-MB-231 cells after P65miRNA-l or P65miRNA-2 transfection were significantly decreased. Conclusion: Targeting silence of P65 expression can inhibit in vitro invasion of TNBC MDA-MB-231 cells, which is related to the down-regulation of MMP-2 and MMP-9 expression and their activities.关键词
三阴性乳腺癌/miRNA/侵袭/P65基因/MMP-2/MMP-9Key words
triple-negative breast cancer/ miRNA/ invasion/ P65 gene/ MMP-2/ MMP-9分类
医药卫生引用本文复制引用
王玲,单保恩,桑梅香,连易水,丁春艳,周欣亮..P65基因对人三阴性乳腺癌细胞MDA-MB-231体外侵袭的影响[J].中国肿瘤生物治疗杂志,2011,18(5):502-507,6.基金项目
河北省科学技术研究与发展计划项目资助(No.10276167). （No.10276167）
