FHIT overexpression in HepG2 hepatoma cells affects growth and cyclin D1 expression in vitro.
FHIT overexpression in HepG2 hepatoma cells affects growth and cyclin D1 expression in vitro.
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HepG2 肝癌细胞中 FHIT 过表达影响体外生长和细胞周期蛋白 D1 表达
DOI：
10.3892/etm.2013.1436
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发表时间：
2014-02
影响因子：
2.7
通讯作者：
Wang L
中科院分区：
文献类型：
--
作者：
Ge J;Shen S;Zhang X;Wang K;Liu B;Sun D;Wang L
本研究旨在探讨脆性组氨酸三联体（FHIT）基因甲基化状态及其对肝癌细胞生长和细胞周期蛋白D1表达的影响。收集人肝癌细胞系HepG 2、Hep 3B和Huh 7的总蛋白，并分析FHIT的表达水平。用甲基化特异性聚合酶链反应（PCR）检测肝癌细胞中FHIT基因启动子区的甲基化状态。随后用5-氮杂-2 ′-脱氧胞苷（5-azorbine）处理HepG 2、Hep 3B和Huh 7细胞，然后检查FHIT表达的恢复。构建β-血凝素（HA）-FHIT质粒，转染HepG 2细胞，观察其对细胞生长的抑制作用。将pHA-FHIT质粒和cyclin D1荧光素酶报告质粒共转染HepG 2细胞，荧光素酶法检测FHIT对cyclin D1转录因子活性的影响。观察到FHIT在Hep 3B和HepG 2细胞中以低水平表达;然而，其在Huh 7细胞中以相对高的水平表达。Hep 3B和HepG 2细胞中FHIT的启动子区被部分甲基化，5-氮杂环丁烷处理诱导FHIT表达增加。FHIT表达增加可抑制HepG 2细胞的生长。pHA-FHIT质粒共转染HepG 2细胞后，cyclin D1启动子的转录活性明显降低，cyclin D1的表达量明显减少。总之，FHIT在HepG 2和Hep 3B肝癌细胞中部分甲基化。过表达FHIT可抑制HepG 2细胞生长，并降低细胞周期蛋白D1的表达。
The aim of this study was to investigate the methylation status of fragile histidine triad (FHIT) and the effects of FHIT on cell growth and cyclin D1 expression in hepatoma cells. The total proteins from the human hepatoma cell lines HepG2, Hep3B and Huh7 were collected and the expression levels of FHIT were analyzed. The methylation status in the promoter region of FHIT in the hepatoma cells was measured using methylation-specific polymerase chain reaction (PCR). The HepG2, Hep3B and Huh7 cells were subsequently treated with 5-aza-2′-deoxycytidine (5-azadc) and the restoration of FHIT expression was then examined. A p-hemagglutinin (HA)-FHIT plasmid was constructed and used to transfect the HepG2 cells, and the inhibitory effects of the transfection on cell growth were then assessed. In addition, HepG2 cells were cotransfected with the pHA-FHIT plasmid and a cyclin D1 luciferase reporter plasmid, and the effects of FHIT on the activity of cyclin D1 transcription factor were analyzed using a luciferase assay. FHIT was observed to be expressed at a low level in Hep3B and HepG2 cells; however, it was expressed at a relatively high level in Huh7 cells. The promoter region of FHIT in the Hep3B and HepG2 cells was partially methylated, and 5-azadc treatment induced an increased expression of FHIT. The increased expression of FHIT inhibited the growth of HepG2 cells. Cotransfection with the pHA-FHIT plasmid significantly inhibited the transcriptional activity of the cyclin D1 promoter and decreased the expression of cyclin D1 in HepG2 cells. In conclusion, FHIT was partially methylated in the HepG2 and Hep3B hepatoma cells. The overexpression of FHIT inhibited cell growth and decreased the expression of cyclin D1 in HepG2 cells.
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影响因子：
6.4
作者：
Wang, Lin;Li, Haiyang;Zhang, Yujing;Santella, Regina M.;Weinstein, I. Bernard
通讯作者：
Weinstein, I. Bernard
影响因子：
25.7
作者：
Behari, Jaideep;Zeng, Gang;Monga, Satdarshan P. S.
通讯作者：
Monga, Satdarshan P. S.
影响因子：
6.1
作者：
Lee, Heun-Sik;Park, Mee-Hee;Yeom, Young Il
通讯作者：
Yeom, Young Il
影响因子：
25.7
作者：
Calvisi, DF;Conner, EA;Thorgeirsson, SS
通讯作者：
Thorgeirsson, SS
Hint1 抑制人结肠癌细胞的生长和激活蛋白 1 活性
影响因子：
11.2
作者：
Wang, Lin;Zhang, Yujing;Weinstein, I. Bernard
通讯作者：
Weinstein, I. Bernard