In vitro and in vivo MMP gene expression localisation by In Situ-RT-PCR in cell culture and paraffin embedded human breast cancer cell line xenografts.
In vitro and in vivo MMP gene expression localisation by In Situ-RT-PCR in cell culture and paraffin embedded human breast cancer cell line xenografts.
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DOI：
10.1186/1471-2407-6-18
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发表时间：
2006-01-24
期刊：
影响因子：
3.8
通讯作者：
Griffiths LR
中科院分区：
文献类型：
--
作者：
Haupt LM;Thompson EW;Trezise AE;Irving RE;Irving MG;Griffiths LR
基质金属蛋白酶（MMP）蛋白酶家族的成员是正常和病理条件下基底膜和细胞外基质降解所必需的。在体外，MT 1-MMP（MMP-14，膜型-1-MMP）的表达在更具侵袭性的人乳腺癌（HBC）细胞系中较高，而在体内，其表达与乳腺肿瘤周围的基质有关。MMP-1（间质胶原酶）与体外MDA-MB-231侵袭相关，而MMP-3（基质溶解素-1）在体内定位于乳腺肿瘤的侵袭细胞周围。由于MMP不储存在细胞内，因此难以将其表达定位于其来源细胞。我们利用独特的原位逆转录-聚合酶链反应（IS-RT-PCR）方法定位在体外和体内的基因表达的MT 1-MMP，MMP-1和MMP-3在人乳腺癌。在体外，在MDA-MB-231和MCF-7 HBC细胞系中检测暴露于伴刀豆球蛋白A（Con A）后的MMP诱导。在体内，我们研究了它们在档案石蜡包埋异种移植物中的表达，这些异种移植物来源于一系列具有不同侵袭性和转移潜力的HBC细胞系。小鼠异种移植物是异质的，含有肿瘤性人类实质以及小鼠基质和血管系统，并提供了与人类疾病状态相关的可重复的体内模型系统。在体外，Con A可增加MDA-MB-231细胞MT 1-MMP基因的表达，降低MCF-7细胞MT 1-MMP基因的表达。MMP-1和MMP-3基因表达在两种细胞系中保持不变。在体内，基质细胞招募到每个异种移植表现出局部MMP基因表达水平的差异。具体地，MDA-MB-231、MDA-MB-435和Hs 578 T HBC细胞系能够影响周围基质中的MMP基因表达。我们已经证明了适用性和敏感性IS-RT-PCR检测MMP基因表达在体外和体内。上皮肿瘤细胞和周围基质细胞中MMP基因表达的诱导与转移潜能的增加相关。我们的数据表明，基质上皮MMP基因表达的贡献，并强调基质金属蛋白酶在乳腺癌的基质-上皮相互作用的复杂性。
Members of the matrix metalloproteinase (MMP) family of proteases are required for the degradation of the basement membrane and extracellular matrix in both normal and pathological conditions. In vitro, MT1-MMP (MMP-14, membrane type-1-MMP) expression is higher in more invasive human breast cancer (HBC) cell lines, whilst in vivo its expression has been associated with the stroma surrounding breast tumours. MMP-1 (interstitial collagenase) has been associated with MDA-MB-231 invasion in vitro, while MMP-3 (stromelysin-1) has been localised around invasive cells of breast tumours in vivo. As MMPs are not stored intracellularly, the ability to localise their expression to their cells of origin is difficult. We utilised the unique in situ-reverse transcription-polymerase chain reaction (IS-RT-PCR) methodology to localise the in vitro and in vivo gene expression of MT1-MMP, MMP-1 and MMP-3 in human breast cancer. In vitro, MMP induction was examined in the MDA-MB-231 and MCF-7 HBC cell lines following exposure to Concanavalin A (Con A). In vivo, we examined their expression in archival paraffin embedded xenografts derived from a range of HBC cell lines of varied invasive and metastatic potential. Mouse xenografts are heterogenous, containing neoplastic human parenchyma with mouse stroma and vasculature and provide a reproducible in vivo model system correlated to the human disease state. In vitro, exposure to Con A increased MT1-MMP gene expression in MDA-MB-231 cells and decreased MT1-MMP gene expression in MCF-7 cells. MMP-1 and MMP-3 gene expression remained unchanged in both cell lines. In vivo, stromal cells recruited into each xenograft demonstrated differences in localised levels of MMP gene expression. Specifically, MDA-MB-231, MDA-MB-435 and Hs578T HBC cell lines are able to influence MMP gene expression in the surrounding stroma. We have demonstrated the applicability and sensitivity of IS-RT-PCR for the examination of MMP gene expression both in vitro and in vivo. Induction of MMP gene expression in both the epithelial tumour cells and surrounding stromal cells is associated with increased metastatic potential. Our data demonstrate the contribution of the stroma to epithelial MMP gene expression, and highlight the complexity of the role of MMPs in the stromal-epithelial interactions within breast carcinoma.
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影响因子：
8.4
作者：
Curran, S;Murray, GI
通讯作者：
Murray, GI
影响因子：
3.8
作者：
Barrett, JM;Puglia, MA;Tozer, RG
通讯作者：
Tozer, RG
影响因子：
6.4
作者：
Bisson, C;Blacher, S;Noel, A
通讯作者：
Noel, A
影响因子：
3.7
作者：
Deryugina, EI;Ratnikov, B;Strongin, AY
通讯作者：
Strongin, AY
DOI：
10.1073/pnas.92.16.7362
发表时间：
1995-08-01
影响因子：
11.1
作者：
BRUNER, KL;RODGERS, WH;OSTEEN, KG
通讯作者：
OSTEEN, KG