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

来源 :南京医科大学学报(自然科学版) | 被引量 : 0次 | 上传用户:whlyxyx
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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。 AIM: To investigate the relationship between hepatocyte nuclear factor 4α (HNF4α) and the expression of angiogenesis-related genes in rat hepatocellular carcinoma (HCC) in vitro and in vivo. Methods: The proliferation of CBRH-7919 cells was induced by overexpression of HNF4α adenovirus. Cell proliferation was assessed by CCK-8 assay. Cell invasion was evaluated by Transwell assay. Angiopoietin-2 (Ang- 2), the expression of vascular endothelial grouth factor (VEGF). In addition, CBRH-7919 cells (blank group) were injected into the residual liver or transfected with the empty vector adenovirus CBRH-7919 cells (negative control group) or transfected with HNF4α overexpressing cells (70%) in vivo. Virus CBRH-7919 cells (experimental group). After 4 weeks, the rats were sacrificed and the expression of Ang-2 and VEGF in the liver specimens was observed by immunohistochemistry. Results: The proliferation and invasion ability of CBRH-7919 cells transfected with HNF4α adenovirus was significantly lower than that of the blank control group (P <0.05). The expression of Ang-2 and VEGF in the experimental group was more than that of the blank control group, Negative control group were significantly inhibited (P <0.05). Immunohistochemical staining of liver tissue in vivo showed that the expression of Ang-2 and VEGF in the experimental group was significantly lower than that in the blank control group (P <0.05). Conclusion: HNF4a can inhibit the proliferation and invasion of hepatocellular carcinoma 7919 and inhibit the expression of Ang-2 and VEGF in hepatocellular carcinoma.
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