Silencing of autocrine motility factor induces mesenchymal-to-epithelial transition and suppression of osteosarcoma pulmonary metastasis.
Silencing of autocrine motility factor induces mesenchymal-to-epithelial transition and suppression of osteosarcoma pulmonary metastasis.
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DOI：
10.1158/0008-5472.can-09-3880
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发表时间：
2010-11-15
期刊：
影响因子：
11.2
通讯作者：
Raz A
中科院分区：
文献类型：
--
作者：
Niinaka Y;Harada K;Fujimuro M;Oda M;Haga A;Hosoki M;Uzawa N;Arai N;Yamaguchi S;Yamashiro M;Raz A
磷酸葡萄糖异构酶（PGI）是一种多功能酶，在细胞内作为糖酵解酶催化葡萄糖和果糖的相互转化，在细胞外作为细胞因子，具有调节肿瘤细胞运动的自分泌运动因子（AMF）等特性。AMF/PGI的过表达可诱导上皮细胞向间质细胞转化（EMT）并增强恶性肿瘤。最近的研究表明，AMF/PGI的沉默导致人肺纤维肉瘤细胞和乳腺癌细胞的间充质向上皮转化（MET），恶性程度降低。在这里，我们构建了一个针对人、小鼠和大鼠AMF/PGI mRNA序列中G390位置的GUC三联体的锤头核酶。用AMF/PGI特异性核酶稳定转染人间充质骨肉瘤MG-63、hs - os -1和小鼠LM8细胞。稳定的转染细胞表现出AMF/PGI表达的有效下调和随后AMF/PGI分泌的消除，导致形态改变，生长、运动和侵袭减少。AMF/PGI沉默诱导MET， E-cadherin和细胞角蛋白上调，vimentin下调。AMF/PGI基因沉默引导下的MET诱导MG-63骨肉瘤最终分化为成熟成骨细胞。此外，MET完全抑制裸鼠LM8细胞的肿瘤生长和肺转移。因此，恶性肿瘤的获得可能部分由AMF/PGI的上调完成，而恶性肿瘤的放弃也可能由AMF/PGI的下调控制。
Phosphoglucose isomerase (PGI) is a multifunctional enzyme that functions in glucose metabolism as a glycolytic enzyme catalyzing an interconversion between glucose and fructose inside the cell, while, PGI acts as cytokine outside the cell, with properties that include autocrine motility factor (AMF) regulating tumor cell motility. Overexpression of AMF/PGI induces epithelial to mesenchymal transition (EMT) with enhanced malignancy. Recent studies have revealed that silencing of AMF/PGI resulted in mesenchymal to epithelial transition (MET) of human lung fibrosarcoma cells and breast cancer cells with reduced malignancy. Here, we constructed a hammerhead ribozyme specific against GUC triplet at the position G390 in the human, mouse, and rat AMF/PGI mRNA sequence. Mesenchymal human osteosarcoma MG-63, H S-Os-1, and murine LM8 cells were stably transfected with the ribozyme specific for AMF/PGI. The stable transfectant cells showed effective down-regulation of AMF/PGI expression and subsequent abrogation of AMF/PGI secretion, which resulted in morphological change with reduced growth, motility, and invasion. Silencing of AMF/PGI induced MET, in which up-regulation of E-cadherin and cytokeratins as well as down-regulation of vimentin were noted. The MET guided by AMF/PGI gene silencing induced osteosarcoma MG-63 to terminally differentiate into mature osteoblasts. Furthermore, The MET completely suppressed tumor growth and pulmonary metastasis of LM8 cells in nude mice. Thus, acquisition of malignancy might be completed in part by up-regulation of AMF/PGI and waiver of malignancy might be also controlled by down-regulation of AMF/PGI.