反义RNA抑制Ezrin表达对乳腺癌细胞侵袭能力的影响

Anti-sense RNA inhibits the expression of Ezrin and invasion of breast cancer cells

目的观察特异性阻断Ezrin的表达对人乳腺癌细胞MDA—MB-231和MCF-7的增殖和侵袭能力的影响。方法将anti—pCR3．1-Ezrin质粒经脂质体介导，转染人人乳腺癌细胞MDA—MB-2316、12和24h，应用Westernblot和逆转录-聚合酶链反应（RT—PCR）方法检测Ezrin的表达变化情况；转染质粒24h后，噻唑蓝（MTF）比色法检测抑制Ezrin对MDA—MB-231和MCF-7细胞体外增殖能力的影响，Boyden小室法检测抑制Ezrin对MDA—MB-231和MCF-7细胞体外侵袭能力的影响。结果转染anti—pCR3．1-Ezrin后，对MDA—MB-231细胞中的Ezrin表达抑制在24h时达高峰。MTY法比色实验结果显示，MDA—MB-231和MCF-7细胞中转染anti—pCR3．1-Ezrin组、转染空质粒组和对照组的A值分别为0．410±0．018、0．765±0．058、0．795±0．061和0．480±0．021、0．632±0．052、0．648±0．059。转染anti—pCR3．1．Ezrin组细胞的增殖受到明显抑制，抑制率分别为（47．9±3．1）％和（32．0±2．8）％（P〈0．05）。Boyden小室法检测结果显示，MDA—MB-231和MCF-7细胞中转染anti—pCR3．1-Ezrin组细胞的侵袭能力分别为对照组的（50．5±3．2）％和（74．8±4．6）％（P〈0．05）。结论Ezrin在乳腺癌的牛长和侵袭过程中发挥蕈要作用。

Objective To observe the influence of proliferation and invasion of human breast cancer cells MDA-MB-231 after inhibition of Ezrin expression by anti-sense RNA. Methods Human breast cancer cell lines MDA-MB-231 and MCF-7 were transfeeted with anti-pCR3. 1-Ezrin with lipofeetamine. Western-blot and RT-PCR were used to detect the expression of Ezrin after transfection for 6, 12, and 24 h, respectively. The proliferative ability of cells was tested by MTT assay, and the metastatic ability of cells was investigated by Boyden cabin after transfection for 24 h. Results After transfection with anti-sense RNA for 24 h, both protein and mRNA expression of Ezrin in MDA-MB-231 cells was significantly reduced at their peak time. After transfection with anti-sense RNA for 24 h, the MTT assay showed that absorbance （A） values in anti-pCR3.1 -Ezrin, vacuity plasmid and blank control groups were 0.410 ± 0.018,0. 765 ±0. 058,0. 795± 0. 061 in MDA-MB-231, and 0. 480± 0. 021,0. 632 ± 0. 052,0. 648 ± 0. 059 in MCF-7, respectively. The results suggested that the proliferation of MDA-MB-231 and MCF-7 cells was inhibited significantly in anti-pCR3.1 -Ezrin group by （47.9± 3.1 ） % and （ 32.0 ±2.8 ） % , respectively. After transfection with anti-sense RNA for 24 h, the invasive ability of MDA-MB-231 and MCF- 7 cells was （ 50.5± 3.2 ） % and （ 74.8 ±4.6 ） % respectively as compared with that of blank control group （ P 〈 0. 05 ）. Conclusion Ezrin plays an important role in the process of proliferation and invasion of breast cancer cells.