层粘连蛋白沉积到IV型胶原上可通过β1整合素增强肝癌细胞的趋触性、化学运动性和黏附性。

Laminin deposition to type IV collagen enhances haptotaxis, chemokinesis, and adhesion of hepatoma cells through beta1-integrins.

作者信息

Torimura T, Ueno T, Kin M, Harad R, Nakamura T, Sakamoto M, Kumashiro R, Yano H, Kojiro M, Sata M

机构信息

The Second Department of Medicine, Kurume University School of Medicine, Japan.

出版信息

J Hepatol. 2001 Aug;35(2):245-53. doi: 10.1016/s0168-8278(01)00127-1.

DOI:10.1016/s0168-8278(01)00127-1

PMID:11580147

Abstract

BACKGROUND/AIMS: In hepatocellular carcinoma, laminin deposition to type IV collagen along the sinusoids is observed with the development of arterial network, coinciding with intrahepatic metastasis. We investigated the influence of laminin deposition to type IV collagen on hepatoma cell adhesion, motility and secretion of matrix metalloproteinases (MMPs), which are indispensable behaviors for tumor metastasis.

METHODS

Hepatoma cell lines (KYN-1, -2 and -3) were used. The expression of integrin subunit mRNAs in hepatoma cells was confirmed by RT-PCR. The influence of laminin addition to type IV collagen on the adhesion, chemokinesis, and migration of KYN-1, -2 and -3 was evaluated by the haptotactic migration, phagokinetic track motility, and cell adhesion assays. The effects of integrin subunits on these activities were evaluated using the function-blocking antibodies for integrins. Phosphorylation of MEK1/2 and secretion of MMPs were investigated by Western blotting and gelatin zymography.

RESULTS

Integrin alpha1, alpha2, alpha3, alpha6 and beta1 subunit mRNAs were detected. The combination of type IV collagen and laminin enhanced the migration, chemokinesis, and adhesion of hepatoma cells compared to that of type IV collagen when used alone. The enhanced activity was significantly suppressed by function-blocking antibodies for integrin alpha1, alpha2, alpha3, alpha6 and beta1 subunits. Hepatoma cells cultured on the combination of type IV collagen and laminin showed phosphorylation of MEK1/2 and increased secretion of MMPs.

CONCLUSIONS

The addition of laminin to type IV collagen enhances hepatoma cell adhesion and motility through beta1-integrins.

摘要

背景/目的：在肝细胞癌中，随着动脉网络的发展，可观察到层粘连蛋白沿肝血窦沉积于IV型胶原，这与肝内转移同时发生。我们研究了层粘连蛋白沉积于IV型胶原对肝癌细胞黏附、运动以及基质金属蛋白酶（MMPs）分泌的影响，这些都是肿瘤转移所必需的行为。

方法

使用肝癌细胞系（KYN-1、-2和-3）。通过逆转录聚合酶链反应（RT-PCR）确认肝癌细胞中整合素亚基mRNA的表达。通过趋触性迁移、吞噬动力学轨迹运动和细胞黏附试验评估层粘连蛋白添加到IV型胶原上对KYN-1、-2和-3细胞黏附、化学运动和迁移的影响。使用整合素功能阻断抗体评估整合素亚基对这些活性的影响。通过蛋白质免疫印迹法和明胶酶谱法研究MEK1/2的磷酸化和MMPs的分泌。

结果

检测到整合素α1、α2、α3、α6和β1亚基mRNA。与单独使用IV型胶原相比，IV型胶原和层粘连蛋白的组合增强了肝癌细胞的迁移、化学运动和黏附。整合素α1、α2、α3、α6和β1亚基的功能阻断抗体显著抑制了增强的活性。在IV型胶原和层粘连蛋白组合上培养的肝癌细胞显示MEK1/2磷酸化和MMPs分泌增加。