Department of Molecular and Cellular Biology, University of Guelph, Guelph, ON N1G 2W1, Canada.
J Cell Sci. 2009 Nov 15;122(Pt 22):4089-98. doi: 10.1242/jcs.052761.
Cellular remodeling of the extracellular matrix (ECM), an essential component of many physiological and pathological processes, is dependent on the trafficking and secretion of matrix metalloproteinases (MMPs). Soluble NSF attachment protein receptor (SNARE)-mediated membrane traffic has documented roles in cell-ECM interactions and the present study specifically examines SNARE function in the trafficking of MMPs during ECM degradation. Using the invasive human fibrosarcoma cell line HT-1080, we demonstrate that a plasma membrane SNARE, SNAP23, and an endosomal v-SNARE, VAMP3 (also known as cellubrevin), partly colocalize with MMP2 and MMP9, and that inhibition of these SNAREs using dominant-negative SNARE mutants impaired secretion of the MMPs. Inhibition of VAMP3, SNAP23 or syntaxin-13 using dominant-negative SNARES, RNA interference or tetanus toxin impaired trafficking of membrane type 1 MMP to the cell surface. Consistent with these observations, we found that blocking the function of these SNAREs reduced the ability of HT-1080 cells to degrade a gelatin substrate in situ and impaired invasion of HT-1080 cells in vitro. The results reveal the importance of VAMP3, syntaxin-13 and SNAP23 in the trafficking of MMP during degradation of ECM substrates and subsequent cellular invasion.
细胞外基质（ECM）的细胞重塑是许多生理和病理过程的重要组成部分，依赖于基质金属蛋白酶（MMPs）的运输和分泌。可溶性 NSF 附着蛋白受体（SNARE）介导的膜运输在细胞-ECM 相互作用中具有明确的作用，本研究特别研究了 SNARE 在 ECM 降解过程中 MMP 运输中的功能。使用侵袭性人纤维肉瘤细胞系 HT-1080，我们证明了质膜 SNARE SNAP23 和内体 v-SNARE VAMP3（也称为细胞溶素）部分与 MMP2 和 MMP9 共定位，并且使用显性负 SNARE 突变体抑制这些 SNARE 会损害 MMP 的分泌。使用显性负 SNARE、RNA 干扰或破伤风毒素抑制 VAMP3、SNAP23 或突触融合蛋白-13 会损害膜型 1 MMP 向细胞表面的运输。与这些观察结果一致，我们发现阻断这些 SNARE 的功能会降低 HT-1080 细胞在原位降解明胶底物的能力，并损害 HT-1080 细胞在体外的侵袭能力。结果表明 VAMP3、突触融合蛋白-13 和 SNAP23 在 ECM 底物降解过程中 MMP 的运输以及随后的细胞侵袭中具有重要作用。
