FIG-ROS融合基因在肝内胆管细胞癌中表达及其意义

目的:探讨FIG-ROS融合基因在肝内胆管细胞癌(ICC)细胞中的表达,以及对其干预后ICC细胞的生物学行为的变化。方法:用Western blot法检测4份不同ICC组织样本及3种ICC细胞株(HUCCT1、REB、QBC939)中ROS蛋白的表达;选择ROS阳性ICC细胞,用一系列表达不同序列ROS-sh RNA与FIG-sh RNA的质粒分别转染该细胞后,用Western blot检测ROS和FIG蛋白表达;选择对ROS和FIG表达抑制作用最强的ROS-sh RNA与FIG-sh RNA序列分别或联合转染上述细胞后,观察细胞增殖、细胞周期、凋亡及集落形成情况。结果:2份ICC组织样本与1个细胞株(HUCCT1)呈ROS阳性表达;转染ROS1-6290 sh RNA和FIG-363 sh RNA对HUCCT1细胞ROS与FIG蛋白表达的抑制作用最强。与未转染的HUCCT1细胞比较,单独转染FIG-363 sh RNA对细胞增殖、凋亡及细胞周期无明显影响(均P0.05),但能明显减少细胞集落形成(P0.05);ROS1-6290 sh RNA单独或联合FIG-363 sh RNA转染均能明显抑制细胞增殖、诱导细胞凋亡与细胞周期阻滞、减少细胞集落形成,且联合转染的效应更为明显(均P0.05)。结论:部分ICC存在FIG-ROS融合基因表达,对两种基因的联合抑制可能是靶向治疗该类ICC的有效途径。

Expression of FIG-ROS fusion gene in intrahepatic cholangiocarcinoma and its significance

Objective: To investigate the expression of FIG-ROS fusion gene in intrahepatic cholangiocarcinoma (ICC) cells and the effects of its intervention on biological behavior of ICC cells. Methods: ROS protein expression in 4 different specimens of ICC tissue and 3 types of ICC cell line (HUCCT1, REB and QBC939) was determined by Western blot analysis; the ROS positive cell line was selected for use and after transfection with a series of plasmids containing different sequences of ROS-shRNAs or FIG-shRNAs respectively, the protein expressions of ROS and FIG in the cells were measured by Western blot analysis. The sequences of ROS-shRNA and FIG-shRNA with highest inhibitory effect on ROS and FIG expression were chosen, which were alone or in combination transfected into the above cells, and after that, the cell proliferation, apoptosis, cell cycle and colony formation were observed. Results: Two specimens of ICC tissue and one ICC cell line (HUCCT1) showed positive ROS expression. Transfection of ROS1-6290 shRNA and FIG-363 shRNA had the most remarkable inhibitory effect on ROS and FIG expression, respectively. Compared with the HUCCT1 cells without any transfection, lone FIG-363 shRNA transfection had no obvious effect on proliferation, apoptosis or cell cycle phase (all P>0.05), but significantly reduced the colony formation of the cells (P<0.05); either ROS1-6290 shRNA transfection alone or in combination with FIG-363 shRNA showed significant effects of suppression of proliferation, induction of apoptosis and cell cycle arrest and inhibition of colony formation, and these effects were more remarkable in cells with combined transfection (all P<0.05). Conclusion: Some kinds of ICC have FIG-ROS fusion gene expression, and the combined inhibition of the two genes may probably provide a hopeful targeted treatment approach for these ICC.