Involvement of hepatocyte growth factor in the development of bone metastasis of a mouse mammary cancer cell line, BALB/c-MC.
Involvement of hepatocyte growth factor in the development of bone metastasis of a mouse mammary cancer cell line, BALB/c-MC.
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DOI：
10.1016/j.bone.2005.12.006
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发表时间：
2006-07
期刊：
影响因子：
4.1
通讯作者：
K. Ono;S. Kamiya;T. Akatsu;Chika Nakamura;Minqi Li;N. Amizuka;Kunio Matsumoto;Toshikazu Nakamura;N. Kugai;S. Wada
中科院分区：
文献类型：
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作者：
K. Ono;S. Kamiya;T. Akatsu;Chika Nakamura;Minqi Li;N. Amizuka;Kunio Matsumoto;Toshikazu Nakamura;N. Kugai;S. Wada
一些癌症经常影响骨骼，骨骼微环境支持某些癌细胞的生长。肿瘤转移到骨后，刺激破骨细胞生成并在骨组织中扩张。肝细胞生长因子(HGF)最初被认为是一种强大的肝细胞有丝分裂原，可促进肿瘤的生长、侵袭和转移。HGF主要由间充质来源的细胞产生，成骨细胞/骨细胞和骨髓基质细胞来源于间充质细胞。然而，目前还不清楚HGF在骨转移中对肿瘤进展有什么影响。在本研究中，我们利用小鼠乳腺癌细胞株BALB/c-MC研究了HGF在骨转移中的作用。注射到小鼠心脏的癌细胞转移到后肢的骨骼。瘤巢周围间质HGF免疫反应阳性，HGF阳性区域可见血管内皮细胞标志CD31表达。为了鉴定产生HGF的细胞，我们测量了培养上清液中HGF的浓度。成骨细胞培养中HGF浓度升高(3.13±0.25 ng/ml)，而BALB/c-MC和骨髓细胞培养中未检测到HGF(0.4 ng/ml)。成骨细胞培养中肝细胞生长因子浓度在10−6M前列腺素E_2作用下增加2.5倍。在BALB/c-MC培养中加入HGF可使细胞数增加一倍。Western印迹分析显示BALB/c-MC表达c-Met/HGF受体。在Matrigel侵袭小室实验中，在底孔中加入HGF可增加BALB/c-MC通过膜侵入底孔的速率。此外，当成骨细胞在底孔中培养时，通过膜侵入底孔的BALB/c-MC细胞数量比没有成骨细胞的检测增加3.7倍。加入HGF抑制剂NK4可完全消除成骨细胞对BALB/c-MC细胞侵袭能力的增强。这些发现表明，成骨细胞产生的HGF诱导癌细胞从肝窦毛细血管向骨髓间隙迁移，并刺激癌细胞在骨微环境中的生长。因此，成骨细胞似乎通过HGF-c-Met信号促进某些癌症的骨转移。
Some cancers frequently affect the skeleton, and the bone microenvironment supports growth of certain cancer cells. After tumors metastasize to bone, they stimulate osteoclastogenesis and expand in the bone tissue. Hepatocyte growth factor (HGF), which was originally identified as a potent mitogen for hepatocytes, promotes tumor growth, invasion and metastasis. HGF is mainly produced by cells of mesenchymal origin, and osteoblasts/osteocytes and bone marrow stromal cells originate from mesenchymal cells. However, it is not clear what effect HGF has on tumor progression in bone metastasis. In the present study, we investigated the roles of HGF in bone metastasis using the mouse mammary cancer cell line BALB/c-MC. Cancer cells injected into hearts of mice metastasized to bone in their hind limbs. HGF immunoreactivity was detected in the stroma surrounding the tumor nests, and blood vessels expressing CD31 (a marker of endothelial cells) were observed in the HGF-positive area. To identify the cells producing HGF, we measured concentration of HGF in culture media. HGF concentration was elevated in osteoblast cultures (3.13 ± 0.25 ng/ml), whereas HGF was undetectable (<0.4 ng/ml) in BALB/c-MC and bone marrow cell cultures. HGF concentration in osteoblast cultures increased 2.5-fold in response to 10−6M PGE2. Addition of HGF to BALB/c-MC cultures caused doubling of the cell number. Moreover, Western blot analysis revealed expression of c-Met/HGF receptor by BALB/c-MC. In the Matrigel invasion chamber assay, addition of HGF to the bottom well increased the rate at which BALB/c-MC invaded the bottom well through the membrane. Furthermore, when osteoblasts were cultured in the bottom well, the number of BALB/c-MC cells that invaded the bottom well through the membrane increased 3.7-fold, compared to assays without osteoblasts. Addition of NK4, an inhibitor of HGF, completely abolished the enhancement of the invasive potential of the BALB/c-MC cells in the presence of osteoblasts. These findings suggest that HGF produced by osteoblasts induces migration of cancer cells from sinusoidal capillaries to bone marrow space and stimulates growth of cancer cells in the bone microenvironment. Thus, osteoblasts appear to promote bone metastasis of some cancers via HGF-c-Met signaling.