抑制MTAl对雌二醇调控ER阳性乳腺癌细胞MMP-9、TIMP-1表达的影响

Study on the role of MAT1 in estradiol regulated MMP- 9, TIMP - 1 expression in ER positive breast cancer

目的：观察转移相关蛋白1（ Metastasis associated protein 1，MTA1）在雌激素调控雌激素受体（Estrogen Recepto，ER）阳性乳腺癌细胞基质金属蛋白酶－9（Matrix metalloproteinase －9，MMP－9）、基质金属蛋白酶组织抑制因子（ Tissue inhibitor of metalprotease －1，TIMP－1）中的可能作用。方法采用慢病毒转染MTA1－shRNA的方法建立特异性抑制MTA1表达的MCF－7模式细胞株。采用10 nM雌二醇（17β－estradiol，E2）处理细胞48 h，Real－time PCR、Western blot分别检测MMP－9、TIMP－1 mRNA与蛋白表达。结果 MTA1－shRNA最大抑制效率为84．9％，提示成功建立了抑制MTA1表达的MCF－7模式细胞株（MCF－7MTA1－shRNA）。 MCF－7野生株（MCF－7WT）在E2处理后MMP－9 mRNA和蛋白表达水平分别上升了46％（P＜0．05）和37％（P＜0．05），TIMP－1 mRNA和蛋白表达水平分别降低了32．3％（P＜0．05）和18．2％（P＜0．05）；相对MCF－7WT，MCF－7MTA1－shRNA MMP－9 mRNA和蛋白表达水平分别降低了42．9％（P＜0．05）和36．7％（P＜0．05），TIMP－1 mRNA和蛋白表达水平未见显著变化；采用E2处理MCF－7MTA1－shRNA后，MMP－9 mRNA和蛋白表达水平未见明显变化，TIMP－1 mRNA和蛋白表达水平分别降低了25．4％（P＜0．05）和32．2％（P＜0．05）。结论 MTA1在雌激素上调ER阳性乳腺癌细胞MMP－9表达的信号转导通路中可能发挥重要作用，但未参与雌激素调控ER阳性乳腺癌细胞TIMP－1表达的信号转导通路。

Obej ctive To investigate the role of metastasis associated protein 1（MTA1）in estrogen reg-ulated expression of matrix metalloproteinase -9（MMP-9）and tissue inhibitor of metalprotease -1（TIMP-1） in estrogen receptor（ ER ） positive breast cancer cells .Methods MTA1 knockdown cell model was generated based on MCF-7breast cancer cell line by transfected with MTA 1-shRNA.The mRNA and protein level of MMP-9 and TIMP-1 in wild type MCF-7（MCF-7WT）and MCF-7MTA1-shRNA before and after 17β-estradiol （ E2） treatment were examined by Real -time PCR and Western blot respectively .Results The MTA1-shRNA showed maximally 84.9%suppression of MTA1 expression in MCF-7,suggesting a satisfied MTA 1 knockdown cell model was established for subsequent experiments .After treated with E2 for 48 h,MCF-7WT showed an incre-ment of 46%（P〈0.05）and 37%（P〈0.05）of the mRNA and protein level of MMP -9 and a decrement of 32.3%（ P〈0.05）and 18.2%（P〈0.05）of TIMP-1;MCF-7MTA1-shRNA showed a decrement of 32.3%（P〈0.05）and 18.2%（P〈0.05）of mRNA and protein expression of MMP -9 respectively but no significant differ-ence in TIMP-1 comparing with MCF-7WT before treated with Estradiol.After E2 treatment,MCF-7MTA1-shRNA didn′t show significant change of MMP -9 except decrements of 32.3%（P〈0.05）and 18.2%（P〈0.05）in the＆amp;nbsp;mRNA and protein levels of TIMP -1.Conclusion MTA1 may be involved in the pathway by which estrogen regulated the expression of MMP -9 but not TIMP-1 in ER positive breast cancer cells .