Laboratory of Cell Transplantation, Institute for Advanced Medical Sciences, Hyogo College of Medicine, Nishinomiya, Hyogo 663-8501, Japan.
Carcinogenesis. 2012 May;33(5):969-75. doi: 10.1093/carcin/bgs109. Epub 2012 Feb 21.
Malignant mesothelioma is an aggressive tumor arising from mesothelial cells of serous membranes. Src family kinases (SFKs) have a pivotal role in cell adhesion, proliferation, survival and apoptosis. Here, we examined the effect of SFK inhibitors in NCI-H2052, ACC-MESO-4 and NCI-H28 cells, mesothelioma cell lines and Met5A, a human non-malignant mesothelial cell line. We found that PP2, a selective SFK inhibitor, inhibited SFK activity and induced apoptosis mediated by caspase-8 in NCI-H28 but not Met5A, NCI-H2052 and ACC-MESO-4 cells. Src, Yes, Fyn and Lyn protein, which are members of the SFK, were expressed in these cell lines, whereas NCI-H28 cells were deficient in Fyn protein. Small interfering RNA (siRNA) targeting Fyn facilitated PP2-induced apoptosis mediated by caspase-8 in NCI-H2052 and ACC-MESO-4 cells. PP2 reduced Lyn protein levels and suppressed SFK activity in all mesothelioma cell lines. Lyn siRNA induced caspase-8 activation and apoptosis in NCI-H28 cells but not in NCI-H2052 and ACC-MESO-4 cells. However, double RNA interference knockdown of Fyn and Lyn induced apoptosis accompanied by caspase-8 activation in NCI-H2052 and ACC-MESO-4 cells. Dasatinib, an inhibitor of multi-tyrosine kinases including SFK, also inhibited SFK activity and induced reduction of Lyn protein levels, caspase-8 activation and apoptosis in NCI-H28 cells but not in other cell lines. Present study suggests that SFK inhibitors induce caspase-8-dependent apoptosis caused by reduction of Lyn protein in Fyn-deficient mesothelioma cells.
恶性间皮瘤是一种源自浆膜间皮细胞的侵袭性肿瘤。Src 家族激酶（SFKs）在细胞黏附、增殖、存活和凋亡中起关键作用。在此，我们研究了 SFK 抑制剂对 NCI-H2052、ACC-MESO-4 和 NCI-H28 细胞（间皮瘤细胞系）和 Met5A（人非恶性间皮细胞系）的影响。我们发现，SFK 选择性抑制剂 PP2 抑制 SFK 活性并诱导 NCI-H28 细胞中 caspase-8 介导的凋亡，但不诱导 Met5A、NCI-H2052 和 ACC-MESO-4 细胞中的凋亡。Src、Yes、Fyn 和 Lyn 蛋白是 SFK 的成员，在这些细胞系中表达，而 NCI-H28 细胞缺乏 Fyn 蛋白。靶向 Fyn 的小干扰 RNA（siRNA）促进了 PP2 诱导的 NCI-H2052 和 ACC-MESO-4 细胞中 caspase-8 介导的凋亡。PP2 降低了所有间皮瘤细胞系中的 Lyn 蛋白水平并抑制了 SFK 活性。Lyn siRNA 诱导 NCI-H28 细胞中 caspase-8 的激活和凋亡，但不诱导 NCI-H2052 和 ACC-MESO-4 细胞中的凋亡。然而，Fyn 和 Lyn 的双重 RNA 干扰敲低诱导了 NCI-H28 细胞中 caspase-8 激活伴随的凋亡，但不诱导 NCI-H2052 和 ACC-MESO-4 细胞中的凋亡。多酪氨酸激酶抑制剂（包括 SFK）抑制剂 dasatinib 也抑制了 SFK 活性，并诱导了 NCI-H28 细胞中 Lyn 蛋白水平的降低、caspase-8 的激活和凋亡，但不诱导其他细胞系中的凋亡。本研究表明，SFK 抑制剂通过降低 Fyn 缺陷型间皮瘤细胞中的 Lyn 蛋白诱导 caspase-8 依赖性凋亡。
