CAJ | 학술논문

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。
목적：관찰전이상관단백1（ Metastasis associated protein 1，MTA1）재자격소조공자격소수체（Estrogen Recepto，ER）양성유선암세포기질금속단백매－9（Matrix metalloproteinase －9，MMP－9）、기질금속단백매조직억제인자（ Tissue inhibitor of metalprotease －1，TIMP－1）중적가능작용。방법채용만병독전염MTA1－shRNA적방법건립특이성억제MTA1표체적MCF－7모식세포주。채용10 nM자이순（17β－estradiol，E2）처리세포48 h，Real－time PCR、Western blot분별검측MMP－9、TIMP－1 mRNA여단백표체。결과 MTA1－shRNA최대억제효솔위84．9％，제시성공건립료억제MTA1표체적MCF－7모식세포주（MCF－7MTA1－shRNA）。 MCF－7야생주（MCF－7WT）재E2처리후MMP－9 mRNA화단백표체수평분별상승료46％（P＜0．05）화37％（P＜0．05），TIMP－1 mRNA화단백표체수평분별강저료32．3％（P＜0．05）화18．2％（P＜0．05）；상대MCF－7WT，MCF－7MTA1－shRNA MMP－9 mRNA화단백표체수평분별강저료42．9％（P＜0．05）화36．7％（P＜0．05），TIMP－1 mRNA화단백표체수평미견현저변화；채용E2처리MCF－7MTA1－shRNA후，MMP－9 mRNA화단백표체수평미견명현변화，TIMP－1 mRNA화단백표체수평분별강저료25．4％（P＜0．05）화32．2％（P＜0．05）。결론 MTA1재자격소상조ER양성유선암세포MMP－9표체적신호전도통로중가능발휘중요작용，단미삼여자격소조공ER양성유선암세포TIMP－1표체적신호전도통로。
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 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 .