Enhanced expression of lumican inhibited the attachment and growth of human embryonic kidney 293 cells
Enhanced expression of lumican inhibited the attachment and growth of human embryonic kidney 293 cells
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DOI：
10.1016/j.yexmp.2010.01.010
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发表时间：
2010-06-01
影响因子：
3.6
通讯作者：
Naito, Zenya
中科院分区：
文献类型：
--
作者：
Ishiwata, Toshiyuki;Yamamoto, Tetsushi;Naito, Zenya
Lumican 是富含亮氨酸的小蛋白聚糖 (SLRP) 家族的成员，它调节各种组织细胞外基质中胶原纤维的组装和直径。据报道，Lumican 在多种肿瘤细胞中表达。 Lumican 可抑制黑色素瘤细胞的生长，但胰腺癌中的 Lumican 与晚期以及腹膜后和十二指肠侵袭相关。在这项研究中，我们阐明了lumican表达的增强是否有助于细胞附着、生长、集落形成、迁移和侵袭。 HEK 293 细胞，稳定转染了 lumican cDNA，在培养基中合成并分泌高水平的 50 kDa lumican 蛋白。细胞呈现出具有长突起的多边形外观，并且细胞与纤连蛋白的粘附程度低于空载体转染的对照细胞（模拟细胞）。相比之下，细胞对 I 型胶原的粘附程度与模拟细胞没有不同。 α5 整合素（纤连蛋白的主要整合素亚基）的表达水平在 Lumican 转染的 HEK 细胞中低于模拟细胞。此外，转染lumican的HEK细胞在体外表现出生长速率降低并且在软琼脂中不形成集落。在 lumican 转染的 HEK 细胞中，AKT、细胞外信号调节激酶 (ERK) 1/2 和哺乳动物雷帕霉素靶蛋白 (mTOR) 的磷酸化降低。在 Lumican 转染的 HEK 细胞和模拟细胞中，细胞迁移和侵袭没有改变。这些发现表明 50 kDa lumican 蛋白在抑制 HEK 细胞附着和生长中发挥重要作用，并且可能抑制整合素途径的激活。 (C) 2010 Elsevier Inc. 保留所有权利。
Lumican is a member of a small leucine-rich proteoglycan (SLRP) family and it regulates the assembly and diameter of collagen fibers in the extracellular matrix of various tissues. Lumican expression was reported in various kinds of tumor cells. Lumican inhibits the growth of melanoma cells, but the lumican in pancreatic cancer correlated with an advanced stage and retroperitoneal and duodenal invasion. In this study, we clarified whether the enhanced expression of lumican contributes to cellular attachment, growth, colony formation, migration and invasion. HEK 293 cell, stably transfected with lumican cDNA synthesized and secreted a 50 kDa lumican protein at high levels in culture medium. The cells showed a polygonal appearance with long projections and the degree of adhesion of the cells to fibronectin was lower than that of empty vector transfected control cells (mock cells). In contrast, the degree of adhesion of the cells to type I collagen was not different from that of mock cells. The expression levels of alpha 5 integrin, the major integrin subunit for fibronectin, were lower in lumican-transfected HEK cells than in mock cells. Furthermore, lumican-transfected HEK cells showed reduced growth rates in vitro and did not form colonies in soft agar. Phosphorylation of AKT, extracellular signal-regulated kinase (ERK) 1/2 and mammalian target of rapamycin (mTOR) decreased in the lumican-transfected HEK cells. Cell migration and invasion were not altered in lumican-transfected HEK cells and mock cells. These findings indicate that the 50 kDa lumican protein plays important roles in the inhibition of HEK cell attachment and growth, and it might inhibit the activation of integrin pathways. (C) 2010 Elsevier Inc. All rights reserved.