基础研究(Basic Research)

FIG-ROS 融合基因在肝内胆管细胞癌中表达及其意义

Published at: 2015年第24卷第2期

朱垒 1 , 黄飞舟 1 , 聂晚频 1 , 邓刚 1 , 黄为 1 , 徐宏博 1 , 易中杰 1 , 郑少鹏 1
1 中南大学湘雅三医院 肝胆胰外科, 湖南 长沙 410013
通讯作者 刚 邓 Email: 2477487669@qq.com
DOI: 10.3978/j.issn.1005-6947.10.3978/j.issn.1005-6947.2015.02.009
基金:
湖南省科学技术厅科技计划资助项目 2014SK3071

摘要

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-shRNA 与FIG-shRNA 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-shRNA 与FIG-shRNA 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 shRNA 和FIG-363 shRNA 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 shRNA 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 shRNA 单独或联合FIG-363 shRNA 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。


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

Abstract

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.


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引用

引用本文: 垒 朱, 飞舟 黄, 晚频 聂, 刚 邓, 为 黄, 宏博 徐, 中杰 易, 少鹏 郑. FIG-ROS 融合基因在肝内胆管细胞癌中表达及其意义[J]. 中国普通外科杂志, 2015, 24(2): 199-205.
Cite this article as: ZHU Lei, HUANG Feizhou, NIE Wanpin, DENG Gang, HUANG Wei, XU Hongbo, YI Zhongjie, ZHENG Shaopeng . Expression of FIG-ROS fusion gene in intrahepatic cholangiocarcinoma and its significance[J]. Chin J Gen Surg, 2015, 24(2): 199-205.