STAT3 transcribes MMP3 and induces MMP3 protein production in HBVEC cells (original) (raw)

2013

Abstract

<p>STAT3 activation of MMP3 has not been previously reported. Therefore, we sought to detect whether STAT3 transcribes MMP3. We stimulated HBVEC cells with Heme, and determined mRNA and protein levels of MMP3 by using qRT-PCR and Western blot. Consistent with the observation that Heme upregulated protein levels of MMP3 as shown in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0071366#pone-0071366-g002" target="_blank">Figure 2A</a>, Heme upregulated mRNA levels of MMP3 in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0071366#pone-0071366-g005" target="_blank">Figure 5A</a>. We then determined whether STAT3 regulates MMP3 using a STAT3-inducible model in HBVEC cells. To this end, HBVEC was transfected with 1 µg of constitutively active STAT3 (caSTAT3), dominant negative STAT3 (dnSTAT3), wild type STAT3 (wtSTAT3) as well as control vector for 24 h. Protein lysates were then made to probe with anti-MMP3 antibody. The results indicate that wtSTAT3 (<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0071366#pone-0071366-g005" target="_blank">Figure 5B</a>) and caSTAT3 increased MMP3 expression (<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0071366#pone-0071366-g005" target="_blank">Figure 5C</a>) whereas dnSTAT3 reduced MMP3 expression (<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0071366#pone-0071366-g005" target="_blank">Figure 5D</a>). When pSTAT3 is reduced by siSTAT3, MMP3 protein expression was inhibited (<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0071366#pone-0071366-g005" target="_blank">Figure 5E</a>).</p

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