论文部分内容阅读
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P>0.05),但能明显减少细胞集落形成(P<0.05);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P<0.05)。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。
OBJECTIVE: To investigate the expression of FIG-ROS fusion gene in intrahepatic cholangiocarcinoma (ICC) cells and the changes of its biological behavior after ICC intervention. Methods: The expression of ROS protein in 4 different ICC tissues and 3 ICC cell lines (HUCCT1, REB, QBC939) was detected by Western blot. ROS-positive ICC cells were selected and expressed in a series of cells expressing different sequences of ROS-sh RNA and FIG. The expression of ROS and FIG protein was detected by Western blot after transfecting the cells with the vector -sh RNA respectively. ROS-sh RNA and FIG-sh RNA sequences with the strongest inhibition of ROS and FIG expression were selected and transfected into these cells respectively After that, cell proliferation, cell cycle, apoptosis and colony formation were observed. Results: Two ICC tissue samples and one HUCCT1 cell line were positive for ROS. The transfection of ROS1-6290 sh RNA and FIG-363 sh RNA had the strongest inhibitory effect on ROS and FIG protein in HUCCT1 cells. Compared with untransfected HUCCT1 cells, FIG-363 sh RNA transfection alone had no effect on cell proliferation, apoptosis and cell cycle (all P> 0.05), but significantly reduced cell colony formation (P <0.05); ROS1 -6290 shRNA alone or in combination with FIG-363 sh RNA transfection could significantly inhibit cell proliferation, induce apoptosis and cell cycle arrest, reduce cell colony formation, and the effect of combined transfection was more obvious (all P <0.05 ). Conclusion: The expression of FIG-ROS fusion gene exists in some ICCs. The combined inhibition of these two genes may be an effective way to target ICC.