肝细胞核因子4α抑制大鼠肝癌细胞增殖、侵袭能力和新生血管生成相关基因表达  

Hepatocyte nuclear factor 4α inhibits the proliferation, invasiveness and expression of new angiogenesis related genes in rat hepatocellular carcinoma cells

作　　者：张勇[1] 吴宁[1] 马永[1] 汪海天[1] 李周骁 卞建民[1] 

机构地区：[1]南京医科大学附属南京医院普外科,江苏南京210006

出　　处：《南京医科大学学报（自然科学版）》2016年第4期430-434,共5页Journal of Nanjing Medical University(Natural Sciences)

基　　金：南京市医学科技发展项目(201308014)

摘　　要：目的 :研究肝细胞核因子4α(hepatocyte nuclear factor 4α,HNF4α)在体内、体外实验中与大鼠肝癌新生血管生成相关基因表达之间的相互关系。方法:将过表达HNF4α腺病毒转染CBRH-7919细胞,采用CCK-8法检测细胞增殖效应,Transwell法检测细胞侵袭能力,RT-PCR和Western blot检测血管生成素2(angiopoietin-2,Ang-2)、血管内皮生长因子(vascular endothelial grouth factor,VEGF)的表达情况。体内试验建立肝大部切除模型(70%),分别在残肝上注入CBRH-7919细胞(空白组)或转染空载体腺病毒CBRH-7919细胞(阴性对照组)或转染过表达HNF4α腺病毒的CBRH-7919细胞(实验组)。4周后处死大鼠,通过免疫组化来观察肝标本Ang-2、VEGF的表达情况。结果:转染过表达HNF4α腺病毒的CBRH-7919细胞增殖能力和侵袭能力较空白、阴性对照组均明显抑制(P<0.05);实验组Ang-2、VEGF在基因、蛋白水平表达较空白、阴性对照组均明显抑制(P<0.05)。体内实验大鼠肝脏组织免疫组织化学切片显示实验组Ang-2、VEGF蛋白表达水平较空白、阴性对照组均明显抑制(P<0.05)。结论 :HNF4a可以抑制大鼠肝癌7919的增殖、侵袭能力,并且抑制肝癌新生血管生成相关基因Ang-2、VEGF的表达。Objective：To investigate the effect of hepatocyte nuclear factor 4α（HNF4α） on the angiogenesis related genes of CBRH-7919 in vitro and in vivo. Methods： CBRH-7919 cells were transfected with HNF4α adenovirus vector. CBRH-7919 cell proliferation was detected by cell counting kit-8（CCK8） assay and the migration and invasion were assessed by Transwell assays.The expressions of Angiopoietin-2（Ang-2） and vascular endothelial growth factor（VEGF） were detected by RT-PCR and Western blotting assay. In vivo, CBRH-7919 cells, CBRH-7919-Null-Vector cells and CBRH-7919-HNF4α cells were respectively injected into the rest of the liver after establishment of subtotal hepatectomy model（70%）. Four weeks later, the expressions of Ang-2 and VEGF in the liver tissues were detected by immunohistochemical assay. Results： In vitro study indicated that after transfected with adenovirus vector LV-HNF4α,the proliferation of CBRH-7919 was significantly inhibited（P〈0.05） and the invasiveness was also significantly inhibited（P〈0.05）compared with the control group,respectively. The result of RT-PCR suggested that the expressions of Ang-2 and VEGF were inhibited after transfected with adenovirus vector. In vivo study showed similar finding that the expressions of Ang-2 and VEGF were inhibited after transfected with adenovirus vector（P〈0.05）. Conclusion： HNF4α can significantly inhibit proliferation and metastasis capability of CBRH-7919, as well as inhibits the expression of Ang-2 and VEGF in CBRH-7919.

关 键 词：肝细胞核因子4Α 肝癌 大鼠肝癌7919 增殖能力 侵袭能力 血管再生 

分 类 号：R735.7[医药卫生—肿瘤]